Ubc 1996-090442

SURIMI-BASED PRODUCT DEVELOPMENT AND VISCOUS
PROPERTIES OF SURIMI PASTE.
by
MOEZ M. BOURAOUI
Engineering Degree, School of Agricultural Engineering, ESIER, Tunisia, 1988

M. A. Sc., University of British Columbia, Canada, 1991
A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF
THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
in
THE FACULTY OF GRADUATE STUDIES
(Department of Chemical Engineering)
We accept this thesis as conforming to the required standard
THE UNIVERSITY OF BRITISH COLUMBIA
December 1995
© Moez M. Bouraoui, 1995

In presenting this thesis in partial fulfilment of the requirements for an advanced
degree at the University of British Columbia, I agree that the Library shall make it
freely available for reference and study. I further agree that permission for extensive
copying of this thesis for scholarly purposes may be granted by the head of my
department or by his or her representatives. It is understood that copying or
publication of this thesis for financial gain shall not be allowed without my written
permission.
Department of CM&MUU £NfyMg£#WQs

The University of British Columbia
Vancouver, Canada
DE-6 (2/88)
Abstract
Raman spectroscopy was used to study the protein structure in raw and
salted surimi from Pacific whiting, and in gels formed by setting (32°C),
cooking (86°C) or setting followed by cooking. The intensity of the peaks
assigned to disulfide bond stretching vibrations increased considerably in the
cooked and set-cooked gels. A smaller increase was found in the gels that
were subject to setting alone. Secondary structure estimation based on the
amide I band indicated a change from predominantly a-helical structure in raw
surimi to similar proportions of a-helical and anti-parallel p-sheet after setting.
A further increase in anti-parallel p-sheet and decrease in a-helix content
occurred during the kamaboko stage. The intensity of C-H stretching
vibrations of the aliphatic residues decreased after salting, setting and
cooking. The set and cooked gel had a better gel strength and fold score
than the cooked gel which, in turn, had better properties than the set gel.
Response surface methodology was used to determine the optimal setting
and cooking time and temperature conditions resulting in a maximized gel
strength, fold score and color, (whiteness index), and a minimized gel
expressible liquid. Cooking temperature was the variable that had the
strongest influence on the gel quality characteristics. Level set programming,
ii
a global optimization method, gave essentially the same results as a gradient
based optimization method. The results obtained by these two methods were
better than those generated by the Simplex technique.
Using a formulation composed of pink salmon surimi, salt, whey protein
concentrate and wheat starch, an optimal final product, kamaboko sausage,
was developed. In addition, when this formulation was applied to herring
surimi, the kamaboko obtained had similar gel strength and elasticity as a
commercial surimi-based product. The effects of frozen storage conditions of
roe herring on its gel making ability (GMA) were investigated. Frozen storage
at around -45°C maintained the GMA for seventy days while storage at -83°C
further increased the GMA period.
The viscous properties of a salmon surimi paste were studied using a
rotational viscometer. The paste behaved as a shear thinning fluid with a
yield stress that increased with temperature up to 21 °C. Viscosity also
increased with temperature up to 21 °C, possibly because of protein-protein
interactions. The Theological data were reasonably well represented by a
simple model which takes into consideration the effects of both shear rate and
temperature.
iii
Table of Contents
Abstract ii
Table of contents iv
List of Tables iv
List of Figures xi
Acknowledgments xiv
Chapter 1: Introduction 1
1.1 Surimi manufacturing 3
1.2 Production of surimi-based products 6
1.3 Freeze denaturation of fish proteins 7
1.3.1 Moisture phase changes 9
1.3.2 Fish lipids 10
1.3.3 Enzymatic denaturation by TMAOase 10
1.4 Thesis outline 11
iv
Chapter 2: Investigation of the Protein Structure of Pacific
Whiting Surimi and Gels Using Raman Spectroscopy 13
2.1 Introduction 13
2.2 Materials and methods 18
2.2.1 Materials 18
2.2.2 Raman spectroscopic analysis 19
2.2.3 Measurement of surimi gel properties 22
2.3 Results and discussion 25
2.3.1 Disulfide (S-S) bond stretching vibrations in the
450-600 cm' region

1
25
2.3.2 Backbone and side chain vibrations in the
610-2000 cm" region1

27
2.3.2.1 The tyrosine doublet near 830 and 850 cm" 1

31
2.3.2.2 Secondary structure estimation from the
amide I and III bands 35
2.3.3 C-H Stretching vibrations in the 2500-3400 cm" region

1
41
2.4 Conclusions 45
Chapter 3: Optimization of the Setting and Cooking Conditions for
Kamaboko Making from Pacific Whiting Surimi 46
3.1 Introduction 46
3.1.1 The Nelder-Mead simplex optimization method 47
v
3.1.2 The Gradient based optimization methods 50
3.1.3 The Level Set Programming (LSP) method 50
3.2.1 Gel strength (compression test) 53
3.2.2 Fold score 53
3.2.3 Gel color test 54
3.2.4 Expressible liquid measurement 54
3.3.1 Curve fitting results 58
3.3.2 Optimization results 65
3.3.2.1 Results of Nelder-Mead Simplex optimization 65
3.3.2.2 Results of NLPQLO Gradient optimization 67
3.3.2.3 Results of LSP optimization 67
3.3.2.4 Multiobjective optimization 70
3.4 Conclusions and recommendations 73
Chapter 4: Kamaboko Production from Roe Herring and Pink
Salmon 75
4.1 Introduction 75
4.2 Objectives 77
4.3.1 Materials 78
vi
4.3.2 General procedures 79
4.3.2.1 Surimi making 79
4.3.2.2 Kamaboko making 80
4.3.2.3 Measurement of surimi gel properties 81
4.3.3 Roe herring samples 81
4.4.1 Frozen storage experiments 82
4.4.2 Optimization experiments 85
4.5 Conclusions 92
Chapter 5: Viscous Properties of Salmon Surimi Paste 94
5.1 Introduction 94
5.4 Conclusions 126
Chapter 6: Summary of Conclusions and Proposed Future Work 128
6.1 Conclusions 128
6.2 Proposed future work 130
Nomenclature 132
vii
Bibliography 136
viii
List of Tables
T a b l e ! Surimi production in four countries, 1985-1991 2
Table 2. Rating system of the fold test 24
Table 3. Tentative assignment of some major bands in the Raman
spectrum of the raw sample (610-2000 cm" ) 1

30
Table 4. Estimated proportions of the buried and the exposed tyrosine
residues 34
Table 5. Secondary structure fractions estimated from the amide I
band by RSAP for the five types of samples 38
Table 6. Experimental design for the optimization of surimi heating
conditions 52
Table 7. Experimental results 56
Table 8. Regression results where "log (G )" is the response

s
variable 60
Table 9. Experimental and estimated values of the 4 response
variables 61
Table 10. Regression results where "F " is the response variable
s 62
Table 11. Regression results where "C" is the response variable 63
Table 12. Regression results where "E" is the response variable 64
Table 13. Optimization of individual response variables using the
Nelder-Mead Simplex method 66
ix
Table 14. Optimization of individual response variables using the
NLPQLO Gradient method 68
Table 15. Optimization of individual response variables using the LSP
method 69
Table 16. Multiobjective optimization results 72
Table 17. Effects of different formulations on surimi gel texture 86
Table 18. Moisture content of different products 91
Table 19. Curve fitting results of the second ISR rheograms 110
Table 20. Curve fitting results of the DSR rheograms 112
Table 21. Results of the curve fitting of the yield stress 119
Table 22. Yield stress values from the first ISR rheograms 120
Table 23. Curve fitting results of Theological data by the ST model 122
x
List of Figures
Figure 1. Flow diagram of commercial surimi manufacturing process 5
Figure 2. Flow diagram of a crab analog production process 8
Figure 3. A schematic layout of a Raman spectrophotometer 20
Figure 4. Example of a force profile recorded by the Instron
instrument 23
Figure 5. Raman spectra of surimi and gels in the 450-600 cm " 1
region showing the S-S stretching band near 530 cm 1

26
Figure 6. Variation with treatment of the normalized intensity of the
530 cm" band

1
28
Figure 7. Raman spectra of raw Pacific whiting surimi in the 610-
2000 cm' region

1
29
Figure 8. Raman spectra of surimi and gels in the 820-912 cm" 1
region, including the 830/850 tyrosine doublet, a) Original
spectra, b) Deconvoluted spectra 32
Figure 9. Raman spectra of surimi and gels in the 1590-1720 cm' 1
region which includes the amide I band, a) Original
region which includes the amide III band, a) Original
xi
region showing the C-H stretching band 42
Figure 12. Variation with treatment of the intensity and of the area of
the -2935 cm" peak

1
44
Figure 13. Variation of roe herring kamaboko gel strength with
frozen storage 84
Figure 14. Combined effects of WPC and WS on salmon surimi gel
strength, 87
Figure 15. Combined effects of WPC and WS on salmon surimi gel
fold score 89
Figure 16. Rheograms of salmon surimi paste at 1 °C 99
Figure 17. Rheograms of salmon surimi paste at 6°C 100
Figure 20. Rheograms of salmon surimi paste at 26°C 103
Figure 21. DSR rheograms 106
Figure 22. Second ISR rheograms 107
Figure 23. DSR rheograms at 1°C 113
Figure 25. DSR rheograms at 11 °C 115
Figure 26. DSR rheograms at 21 °C 116
xii
Figure 28. DSR rheograms and the predictions of the ST model 124
Figure 29. Second ISR rheograms and the predictions of the ST 125
model
xiii
Acknowledgments
I would like to express my most sincere gratitude to my supervisors, Dr. Bruce
Bowen and Dr. Shuryo Nakai, for their invaluable supervision, help and
understanding. I greatly appreciate their contributions and their ideas. My
special thanks go to Dr. Tim Durance and Dr. Ken Pinder, my committee
members, for their contributions to this project and their guidance. I also
thank Dr. Eunice Li-Chan for her great help with the Raman spectroscopy, Dr.
Jaouad Fichtali for his contributions to this project and Mr. Dragan Macura for
his help with the IRAP project.
I extend my thanks to everyone affiliated with the Department of Chemical
Engineering and the Department of Food Science, and to all my friends in
Canada and Tunisia.
I am unable to sufficiently thank my parents, Mansour and Radhia, my dear
wife, Amna, my sister, Hella, and all my family members for their great love,
their sacrifices and their encouragements.
xiv
Chapter 1. Introduction
Chapter 1
Introduction
Surimi is a Japanese term for the wet protein concentrate of mechanically
deboned and water-washed fish flesh (Okada, 1992; Lee, 1984). It is often
mixed with cryoprotectants to maintain the functional properties of its
myofibrillar proteins. The high concentration of these proteins produces,
upon cooking, an elastic and chewy texture that can be made to resemble that
of shellfish (Lee, 1984). For centuries, the Japanese have been producing
surimi mainly from Alaska pollock. Around the twelfth century, Japanese
fishermen discovered that when surimi was salted, ground and steam or broil
cooked, the end product, called kamaboko, could be kept longer (Lee, 1984).
Surimi technology has evolved to produce such shellfish analogs as crab and
shrimp. Recently, the USA has become interested in this technology
especially since it is the major producer of Alaska Pollock. Table 1 shows the
trends in surimi production in several countries (Sproul and Queirolo, 1994).
The table demonstrates that surimi production in the USA jumped from none
in 1985 to 160,000 metric tons in 1991. Other countries such as Canada,
Norway and the UK have also started to produce surimi from under-utilized
1
Table 1. Surimi production in four countries, 1985-1991
Production (1,0001)
Year Japan USA Korea Thailand
1985 490 0 20 10
1987 480 20 30 20
1989 390 80 30 20
1991 190 160 10 20
2
fish species (Hastings and Currall, 1989). In Japan, apart from pollock, other
fish species are being used for surimi production. These species include
sardine mackerel, atka mackerel and horse mackerel. Surimi of good quality
can also be made from New Zealand hoki (Macruronus novaezelandiae). In
1988, New Zealand produced 22,800 metric tons of surimi from this species
(Holmes et al., 1992). Pacific whiting is another under-utilized species that
has recently been used for surimi production. It is very abundant on the West
Coast of North America from British Columbia to California (Ohshima et al.,
1993). The allowable catch of this species in the USA ranges between
140,000 and 250,000 metric tons per year (Yongsawatdigul et al., 1995).
These trends should dramatically increase the amount of fish products
available to humans; about half of the harvested fish in the world in 1986 was
reduced to animal feed (Pigott, 1986). Because of the high nutritional and
functional properties of surimi, extensive research is needed to further
develop and improve the production of shellfish analogs as well as other non-
seafood imitations from this important protein source.
1.1 Surimi manufacturing
Because Alaska Pollock is highly susceptible to freeze denaturation
(Matsumoto and Noguchi, 1992), surimi used to be produced and consumed
on a same-day basis. In 1959, Nishiya's research team discovered that by
3
washing out the water soluble components from the minced fish and adding
cryoprotectants such as sugar compounds and polyphosphates, the functional
properties of surimi could be maintained longer during frozen storage (Okada,
1992) . This was a threshold for the production of frozen surimi. The main
cryoprotectants used are sucrose and sorbitol. Their presence, at low
concentrations (i.e., 4% by weight) increases the amount of bound water
which decreases the displacement of water from protein surfaces and hence
reduces protein freeze denaturation (Arakawa and Timascheff, 1982;
Matsumoto and Noguchi, 1992).
Figure 1 summarizes the overall process of commercial surimi production.
The process starts with deboning and gutting to remove the head, viscera and
most of the backbone. The product is then minced and washed with water.
Washing is very important because it removes most of the sarcoplasmic
proteins, inorganic salts, and other undesirable water soluble compounds
such as blood. It is usually carried out in three to four cycles (Ohshima et al.,
1993) . Refining is then performed to remove any connective tissues, skin
scales, small bones, etc. The soft meat is selectively forced through the
perforations of a cylindrical screen [perforation size varies between 1 and 3
mm (Toyoda et al., 1992)]. The excess water due to the washing process is
then eliminated by a screw press. This dewatering reduces the moisture
content of the refined mince from 90% to around 80% (wet basis). The
4
raw fish (100%)
deboning and gutting
\1/
mincing
washing with water
refining
dewa ering
blending of additives
(i.e., cryoprotectants)
filling and packaging
freezing
frozen surimi (20 to 30 %)
Figure 1. Flow diagram of commercial surimi manufacturing process
5
product is then mixed with cryoprotectants in a silent cutter. Then, the surimi
is extruded as blocks (i.e., 10 kg) and packed in polyethylene bags. Surimi
blocks are then frozen at -25°C or lower (Toyoda et al., 1992). The surimi
yield is between 20 and 30% (raw material weight basis); it varies with the fish
species, size and season as well as the manufacturing equipment used
(Toyoda et al., 1992). Surimi is popular because of its high nutritive value,
high protein content and low fat. For example, surimi produced by Alaska
Pacific Seafoods (Kodiac, Alaska) from Alaska pollock contained 75.3%
water, 15.4% protein, 4% sorbitol, 4% sucrose, 0.3% sodium
tripolyphosphate, and 1.0% fat and ash (Nicklason and Pigott, 1989).
1.2 Production of surimi-based products
Most of surimi-based products are gelled surimi. The Japanese have been
producing surimi-based products for centuries. The washed fish flesh, when
combined with certain ingredients, mixed and kneaded, and steamed, forms a
fish gel called kamaboko (Pigott, 1986). To name but a few, there is steamed
kamaboko, broiled kamaboko and fried kamaboko (Pigott, 1986).
More recently, shellfish analogs have been produced from surimi. These
include imitations of crab, lobster, scallop and shrimp (Lee, 1984). Crab
imitation has gained popularity not only in Japan and Korea but also in the
6
USA. For instance, because of overfishing, the US production of King Crab
plunged from 185 million pounds in 1980 to less than 40 million pounds in
1982 (Gwinn, 1992). It has continued to fall further in succeeding years. This
shortage of crab meat, along with its high price, has created a significant
market for the production of crab analogs. A crab analog manufacturing
process is summarized in Figure 2 (Lee, 1984; Wu, 1992). Surimi is also
used in such products as soups, meat pies and fish cakes.
Lee et al. (1992b) stated that the texture of surimi-based products composed
only of surimi and salt is inclined to be rubbery. To improve the texture
quality and water binding capacity as well as the product stability for frozen
distribution, other ingredients are added. These include starch (i.e., wheat or
potato starch), egg white, etc., which become entrapped within the gel matrix
thereby improving its strength and texture (Lee et al., 1992b). Modified starch
was reported to improve the gel freeze-thaw stability (Lee et al., 1992b).
Other ingredients may also be added depending on the final product desired.
These include oil, flavorings and colorants, to name a few.
1.3 Freeze denaturation of fish proteins
Frozen storage of fish products is widely used by the industry to prolong the
storage life of seafood products. However, extended storage causes the
7
tempering to -8 to -4°C
comminution with other ingredients

(i.e., salt, water, starch, flavor)
pumping and sheet forming (extrusion)
setting and cooking
slitting
bundling
coloring
wrapping
4
cut mg
packaging
I
pasteurization
si,
coo mg
freezing
Figure 2. Flow diagram of a crab analog production process
8
deterioration of the texture, the flavor and the color of the fish. Several
mechanisms have been postulated to explain the causes of the loss of fish
quality. Some of these mechanisms are discussed below.
1.3.1 Moisture phase changes
The formation of ice crystals could cause damage to fish proteins. These ice
crystals are larger when the product is frozen at slow freezing rates or when
the storage temperature fluctuates (Wheaton and Lawson, 1985). Ice crystals
can break cells, rupture membranes and cause drip loss upon thawing of the
fish product (Shenouda, 1980). It is, therefore, recommended to freeze the
product at fast rates allowing the passage of every part of the product through
the critical freezing zone (-0.8 to -5°C) within five to ten hours (Wheaton and
Lawson, 1985).
Matsumoto (1980) and Matsumoto and Noguchi (1992) explained that the
migration of water molecules to form ice crystals causes the dehydration of
the protein molecules. This phenomenon triggers the denaturation of the fish
myofibrillar proteins through aggregation and unfolding. A multitude of
cryoprotectants such as sucrose, sorbitol, phosphates, can be used to reduce
freeze denaturation during frozen storage (Matsumoto and Noguchi, 1992).
9
As water in the fish freezes out, the salt concentration increases. This
increase can cause protein aggregation. Connel (1964) reported that a salt
concentration of 10% caused the most protein damage. This damage is
reported to mainly affect the myosin proteins (Shenouda, 1980).
1.3.2 Fish lipids
Dehydration (caused by freezing) as well as exposure to oxygen (i.e., due to
poor packaging) can increase lipid oxidation during frozen storage of fish.
Shenouda (1980) reported that the oxidized products of lipids interact with the
proteins' functional groups decreasing the solubility of the proteins. It also
causes rancidity and an undesirable fishy taste. Lipid oxidation increases
with storage time and with the fat content of the fish.
1.3.3 Enzymatic denaturation by TMAOase
During the frozen storage of some fish species, especially the gadoid family
(cod, pollock, whiting, hake, etc.), an enzymatic reaction causes the
degradation of trimethylamine oxide (TMAO) into dimethylamine (DMA) and
formaldehyde (FrHO) (Shenouda, 1980). This reaction, caused by the
TMAOase enzyme, is slowed at very low storage temperatures (Wheaton and
Lawson, 1985). FrHO causes cross-linking of proteins and decreases their
10
extractabilty. It mainly affects the tropomyosin and the heavy chains of
myosin (Childs, 1973).
There are still many uncertainties regarding the mechanisms responsible for
fish protein denaturation during frozen storage, and especially concerning the
interactions between the different mechanisms, their relative importance and
the stages at which they occur.
1.4 Thesis outline
This thesis was motivated by the need to provide alternative value-added fish
products from under-utilized fish species. It consists of a comprehensive
study of the various processing factors that play a role in the overall surimi gel
production cycle.
In Chapter 2, the mechanisms of Pacific whiting surimi gelation were studied
using the Raman spectroscopy technique. This method makes it possible to
study the proteins in intact pastes and gels without having to isolate or extract
the myofibrillar proteins.
The effects of setting and cooking conditions on the quality of the final
product (Pacific whiting surimi gel) are studied in Chapter 3. The optimal
11
setting and cooking temperatures and times were determined by finding the
conditions that maximize three and minimize one gel texture quality variable.
Surimi production from two under-utilized species, pink salmon and roe
herring, is investigated in Chapter 4. The effects of frozen storage conditions
on the gel making ability as well as the effects of gel improving ingredients on
the gel texture are discussed. An optimal formulation for the production of
kamaboko sausage was developed.
The viscous properties of a surimi paste consisting of the optimal formulation
obtained in Chapter 4 are studied in Chapter 5.
12
Chapter 2. Raman spectroscopy of surimi
Chapter 2
Investigation of the Protein Structure of Pacific

Whiting Surimi and Gels Using Raman Spectroscopy
2.1 Introduction
Gelation is one of the most important functional properties of surimi. Gelation
is accomplished by first grinding surimi with salt to increase the solubility or
the extractability of the myofibrillar proteins. The resulting paste is then
slowly set at temperatures below 50°C before being cooked at higher
temperatures (usually above 70°C). The formed gel has an elastic and chewy
texture that can be made to resemble the texture of shellfish flesh (Lee,
1984).
A thorough understanding of the mechanisms that govern surimi gelation
would be useful for the improvement of gel quality and texture. Niwa (1992)
indicated that grinding surimi with salt causes the formation of actomyosin
from myofibrils. Myosin has a molecular weight of roughly 480,00 to 500,000
and contains over 40 sulfhydryl residues but no disulfide bonds (Nakai and Li-
Chan, 1988). Actin has a molecular weight of about 42,000 to 47,000 and
contains between 8 and 13 sulfhydryl residues but no disulfide bonds (Nakai
and Li-Chan, 1988). During the setting stage {suwari in Japanese), Gill and
13
Conway (1989) found that heavy meromyosin (HMM S-2) and light
meromyosin (LMM) fragments of the cod myosin tail region were inaccessible
to chymotryptic digestion. They assumed that tail-tail interactions took place.
When isolated, only the LMM fragments formed a gel (Sano et al., 1990).
Hydrophobic interactions have been proposed to be the most dominant
mechanism for the formation of a three-dimensional gel network (Chan et al.,
1992; Niwa, 1992; Stone and Stanley, 1992). Many studies have
demonstrated the importance of hydrophobic interactions in isolated myosin
or actin fractions, actomyosin, meat sols or salt extracts (Nakai and Li-Chan,
1988). In the presence of a hydrophobic probe, sodium anilino-naphthalene
8-sulfonate (ANS), the fluorometric intensity of actomyosin solutions from
setting flatfish increased with heating (Niwa, 1992). During cooking
(kamaboko making or gel strengthening), as more protein unfolding takes
place, interactions between exposed hydrophobic sites become stronger. In
addition, Itoh et al. (1979) postulated an increase in disulfide bonds, based on
a decrease in the SH content of carp actomyosin solution with increasing
temperatures up to 80°C. Niwa (1992) also proposed that some disulfide
bonds may be formed due to oxidation of sulfhydryl residues. Other
researchers, including Tsukamasa et al. (1993), have suggested that enzyme
(transglutaminase) catalyzed cross-linking may occur during setting.
14
In most of the reported literature, the mechanisms of protein sol-to-gel
transition have been postulated by studying the protein structure after
dissolving the gel. A great deal of uncertainty, therefore, still exists regarding
the exact chemistry of surimi gelation. Research is needed to elucidate the
mechanisms involved at each stage of gelation.
Protein molecules are polymers composed of twenty different amino acids
linked together by peptide bonds. The function of each protein depends on its
three-dimensional structure (Branden and Tooze, 1991). Due to the
difficulties in determining a particular protein's tertiary structure, the
knowledge of its secondary structure can be used to predict its overall three-
dimensional structure and consequently the protein's function (Yada et al.,
1988). The secondary structure is usually classified as a-helix, p-sheet or
random coil. Branden and Tooze (1991) explained that, for the a-helices, the
cp and \\t angles for the consecutive residues are around -60° and -50°,
respectively. (q> is the angle of rotation around the N-Ca bond, whereas vy is
the angle around the C a - C bond from the same Ca-atom.) An a-helix has 3.6
residues per turn. On the other hand, p-sheets are composed of several
regions, p-strands, of the polypeptide chain. Each strand contains from five to
ten residues. The strands are aligned adjacent to each other. The p-sheet is
called parallel when the amino acids in the aligned p-strands run in the same
biochemical direction, amino terminal to carboxy terminal. When the amino
15
acids in successive strands run alternatively once from the amino terminal to
the carboxy terminal and once from the carboxy terminal to the amino
terminal, the sheet is called antiparallel (Branden and Tooze, 1991).
Raman spectroscopy is a vibrational spectroscopic technique which can be
used to monitor changes in chemical structure and environmental changes
around atoms. The Raman scattering phenomenon which results in shifts in
wavelength of an exciting incident beam is due to inelastic collisions of the
incident photons with sample molecules. This technique is distinguished by
its ability to analyze solids as well as solutions, and therefore, has great
potential to study food systems (Li-Chan et al., 1994). Raman spectroscopy
has been used to study the myosin substructure (Carew and Asher, 1975) and
the effects of inorganic salts on myosin solutions (Barrett and Peticolas,
1978). Caille et al. (1983) reported on the effects of Mg , ATP and C a

2+ 2+
on
the Raman spectra of intact muscle fibers and internally perfused fibers.
Raman spectroscopy was successfully employed to study the gelation of
lysozyme (Li-Chan and Nakai, 1991) and the gelation of whey proteins
(Nonakaetal., 1993).
Since the Raman spectra of macromolecules such as proteins often consist of
broad bands from overlapping components, many attempts have been made
to apply curve^fitting or mathematical deconvolution techniques to aid in the
16
interpretation of the spectral data. Deconvolution is the unraveling of a band
containing overlapping peaks into separate peaks. This method is utilized in
many areas of science and engineering (i.e., telephone communication,
seismology, etc.) where spectral peaks are overlapping and are therefore
difficult to interpret. Deconvolution was successfully used by Luu et al. (1982)
to study solvent-solute interactions in aqueous solutions, while Mathlouthi and
Portmann (1990) used deconvolution to compare Raman spectra of water with
aqueous solutions of sweeteners. While Fourier deconvolution has been the
most commonly used method, maximum-likelihood spectral restoration or
deconvolution has recently been reported to give superior resolution
enhancement (deNoyer and Dodd, 1991). Unlike Fourier deconvolution which
ignores noise, maximum-likelihood deconvolution takes noise into
consideration. The maximum-likelihood deconvolution requires (Mendel,
1990):
• the specification of a probability model for the measured output;
• the determination of a formula for the likelihood function; and,
• the maximization of the likelihood function.
For noise with a normal distribution, the likelihood function to be maximized is
(deNoyer and Dodd, 1991):
17
P=l 1 u exp[- —j ]
1 = i -42noi 2<jt
where {y y
1( 2l y„} is a measured data set, {CH, O ,2 o } is a more resolved
n
spectrum, ® denotes convolution, a is the standard deviation and s is the
peak shape function. To our knowledge, these techniques have not been
applied before to the study offish myofibrillar proteins.
The objective of this study was to investigate the mechanisms of Pacific
whiting surimi gelation under different processing conditions using Raman
spectroscopy, a method that offers the major advantage of being able to study
changes in protein structure in intact pastes and gels, without having to
isolate or extract the myofibrillar proteins.
2.2 Materials and methods
2.2.1 Materials
Frozen surimi made from Pacific whiting was donated by Ucluelet Seafood
Processors Ltd. (Ucluelet, B.C., Canada). Five types of samples were
studied: raw surimi, salted surimi (ground with 3% salt in a vacuum cutter
18
[Stephan, Model VCM-5, Columbus, OH]), set gel (32°C for 19 min), cooked
gel (86°C for 12 min) and set-cooked gel (setting at 32°C for 19 min followed
by cooking at 86°C for 12 min). A more detailed description of the procedures
used for preparing surimi gels is given in section 4.3.2.2.
2.2.2 Raman spectroscopic analysis
The Raman spectrum of each sample was measured on a Jasco Model NR-
1100 laser Raman spectrophotometer (Jasco Inc., Tokyo, Japan) with
excitation from the 488-nm line of a Spectra-Physics Model 168B argon ion
laser (Spectra-Physics, Mountain View, CA). A schematic layout of the
Raman spectrophotometer used is shown in Figure 3. Laser light is focused
on the sample. The resulting scattered light is accumulated by tollection
optics" and directed to a double monochromator. The latter instrument
separates the scattered light depending on its frequency. Then, the Raman
spectrum is detected before being stored in a personal computer interfaced to
the instrument (Li-Chan et al., 1994). The Raman scattering of samples
placed in hematocrit capillary tubes in a transverse/transverse arrangement
(capillary held horizontally and incident laser beam perpendicular to the
capillary axis) was measured at ambient temperature under the following
conditions: laser power, 100 mW; slit height, 4 mm; spectra resolution, 5 cm"1
19
Collection
optics
Laser 1—H Sample

o Double Monochromator
Detector
Computer
Figure 3. A schematic layout of a Raman spectrophotometer
20
at 19,000 cm" ; sampling speed, 120 cm" min" with data collected at every
1 1 1
cm" ; 6 to 10 scans averaged per sample. These conditions were based on

1
the recommendations of Nonaka et al. (1993), while taking into consideration
the nature of our samples (surimi paste and gels). Duplicate samples were
analyzed in all cases.
Spectral smoothing (to improve the signal to noise ratio), baseline correction,
normalization against the C H bending vibration at -1450 cm" , maximum-

2
1
likelihood spectral restoration or deconvolution (to unravel overlapping
peaks), integration (to determine peak areas) and other computations were
performed using LabCalc (Galactic Industries Corp., Salem, NH) with Square
Tools (Spectrum Square Associates, Ithaca, NY) software on an IBM
compatible (486DX-33) personal computer. Estimation of the secondary
structure composition of the samples based on the Raman spectra in the
amide I region was carried out using the Raman spectral analysis package
(RSAP) of Przybycien and Bailey (1989), which is based on the algorithms of
Williams (1983) for least-squares analysis of the amide I band. Assignments
of some peaks in the Raman spectra to specific vibrational modes of amino
acid side chains or the polypeptide backbone were made according to
published literature (Tu, 1986; Colthup et al., 1990; Li-Chan et al., 1994).
21
2.2.3 Measurement of surimi gel properties
The quality of the surimi gels obtained after setting alone, cooking alone, and
setting followed by cooking needs to be studied in order to try to correlate it to
the Raman spectroscopy results. The surimi gel quality tests performed were
the compression test and the fold test. The compression test is an indicator
of the gel cohesiveness. It was carried out using an Instron Universal Testing
Instrument (Model 1122, Canton, Massachusetts). The test consisted of
compressing a surimi gel cylinder, having a diameter of 30 mm and a height
of 20 mm, to 90% deformation at a cross head speed of 20 mm/min. The
variation of force with time was recorded. The gel strength was the product of
the force and the distance at the point of rupture (Lee, 1984) (see Figure 4).
The distance at the point of rupture was calculated by multiplying the time of
rupture by the cross head speed. The fold test indicates gel elasticity. It was
conducted by folding a 3 mm thick slice of surimi gel (diameter = 30 mm)
slowly in half lengthwise and then in half again while examining it for
structural failure. A rating score from 1 to 6 was used. The rating system is
shown in Table 2 (Lanier, 1992).
22
Figure 4. Example of a force profile recorded by the Instron instrument
23
C h a p t e r 2. R a m a n spectroscopy o f surimi
Table 2. Rating system of the fold test
Fold score Gel condition
6 No crack showing after folding twice
5 No crack showing after folding in half
4 Cracks gradually when folded in half
3 Cracks immediately when folded in half
2 Does not break by finger pressure
1 Breaks by finger pressure
24
2.3 Results and discussion
The set gel had a gel strength of 246 ± 35 N.mm (n=4) and a fold score of 3
while the cooked sample had a gel strength of 2972 + 87 N.mm (n=4) and a
fold score of 4. The set and then cooked gel had the best properties
consisting of a gel strength of 3897 + 104 N.mm (n=4) and a fold score of 6.
Raman spectra were measured in three wavenumber regions for each
sample. These regions were 450-600 cm' (to assess S-S stretching of
1
disulfide bonds), 610-2000 cm" (to estimate secondary structure and various
1
side chain vibrations) and 2500-3400 cm" (to evaluate C-H stretching
1
vibrations of aliphatic residues). The Raman spectra were reasonably
reproducible.
2.3.1 Disulfide (S-S) bond stretching vibrations in the 450-600 cm' 1
region
Figure 5 shows the Raman spectra of the five types of samples studied. A
major peak was found at 529 cm" (raw, salted and set samples) or at 531 cm"
1 1
(cooked and set-cooked samples), assigned to the S-S stretching vibrations
of disulfide bonds in a gauche-gauche-trans conformation (Li-Chan et al.,
25
Wavenumber ( c m " ) 1
Figure 5. Raman spectra of surimi and gels in the 450-600 cm " 1

region
showing the S-S stretching band near 530 cm" 1
26
1994). Although the normalized intensity of this band increased only slightly
after salting and setting, a considerable increase was observed in the cooked
gel and especially in the set-cooked gel (Figure 6). This may be attributed to
either an increase in the number of disulfide bonds or a change in the
environment around existing disulfide bonds during the gel strengthening
stage at high temperatures. Increase in the intensity of the Raman band
around 528 cm" was previously observed upon gelation of hen egg white
1
lysozyme at high temperature (Li-Chan and Nakai, 1991), whereas thermally-
induced gelation of whey proteins resulted in decreased intensity of the S-S
stretching bands near 508 cm" (Nonaka et al., 1993). An increase in disulfide
1
bonds due to gel strengthening of fish proteins was proposed by Itoh's group
(1979) who found that the sulfhydryl content of carp actomyosin solution
decreased with increasing temperature. Therefore, it is thought that the
cooking treatment caused a considerable increase in the number of disulfide
bonds of the surimi gels of this study.
2.3.2 Backbone and side chain vibrations in the 610-2000 cm' region 1
The Raman spectrum of raw surimi in this region is shown in Figure 7. A
tentative assignment of the major peaks of this spectrum to vibrational modes
of the amino acid side chains or peptide backbone is depicted in Table 3.
Further detailed analysis of selected bands in this region was carried out.
27
2.5
Raw Salt Set Cook Set & cook
Treatment
Figure 6. Variation with treatment of the normalized intensity of the 530 cm

band
28
CO
to
CD
Wavenumber ( c m 1
)
Figure 7. Raman spectra of raw Pacific whiting surimi in the 610-2000 cm

region
29
Table 3. Tentative assignment of some major bands in the Raman
spectrum of the raw sample (610-2000 cm" ) 1
wavenumber (cm" ) 1
assignment
761 Trp
834, 850 Tyr
879 Trp
940 a-helix
1005 Phe
1062 p-sheet
1128 Backbone C-N stretch
1213 Tyr, Phe
1254 Amide III
1303 CH bending or amide III (a-helix)
1322, 1342 Trp
1403 Asp, Glu C O O "
1450 C H bending
2
1658 Amide I
30
2.3.2.1 The tyrosine doublet near 830 and 850 cm" 1
The original and the deconvoluted Raman spectra in the wavenumber region
from 820 to 912 cm' for the five types of samples are shown in Figures 8a
1
and 8b, respectively. Special attention was given to the bands located near
830 and 850 cm' 1

because of their usefulness in monitoring the
microenvironment around tyrosyl residues (Li-Chan et al., 1994). This
tyrosine doublet band arises from vibrations of the para-substituted benzene
ring which are affected by the environment and the involvement of the
phenolic hydroxyl group in hydrogen bonding. In the case of tyrosine
residues which are exposed to the aqueous or polar environment and act as
simultaneous acceptor and donor of moderate to weak hydrogen bonds, the
intensity ratio of the doublet bands (Uso/sa)) usually ranges from 0.90-1.45, but
can be as high as 2.5. On the other hand, U50/830 values for tyrosine residues
which are buried in a hydrophobic environment and which tend to act as
hydrogen donors usually range between 0.7 and 1.0, and can be as low as
0.3 in the case of extremely strong hydrogen bonding to a negative acceptor
(Tu, 1982; Li-Chan et al., 1994).
The intensity ratio ISSO/KJO of raw and salted surimi were 1.18 and 1.14,
respectively suggesting that the tyrosine residues of these samples were
mainly exposed. The ISSO/KJO ratios decreased to 0.59 and 0.22 for the set and
31
gOO 860 820 890 870 850 830
Wavenumber (cm ) Wavenumber (cm ) 1
Figure 8. Raman spectra of surimi and gels in the 820-912 cm" region,
1
including the 830/850 tyrosine doublet, a) Original spectra, b) Deconvoluted

spectra
32
C h a p t e r 2 . R a m a n spectroscopy o f surimi
set-cooked samples, respectively. This suggests that some tyrosine residues
became buried in a more hydrophobic environment and were involved as
strong hydrogen bond donors after these samples were processed by setting
as well as setting followed by cooking. On the other hand, the sample which
received only the cook treatment had a high USO/KJO ratio of 3.1, indicating that
some of the tyrosine residues became exposed to a more polar environment
and were involved as strong hydrogen bond acceptors on the surface of the
protein molecule. Tu (1986) developed an equation for estimating the
proportions of the buried and the exposed tyrosine residues. The equation is:
0.5 N buried
+ 1.25N exposed ~ '850/830
where N is the mole fraction (N b uhed +N exposed = 1). The numerical constants
in this equation were obtained by fitting the general form to the X-ray
diffraction data of Kannan et al. (1975). Table 4 lists the proportions of the
buried and the exposed tyrosine residues (TR) estimated by Tu's equation.
This table shows that the TR of the raw and the salted surimi were mainly
exposed. Setting caused most of the TR to become buried. The negative
numbers shown for the cooked and the set-cooked samples show the
limitations of Tu's equations at too high or too low values of USO/KIO- However,
these numbers might be an indication that all the TR of the cooked sample
were exposed whereas all the TR of the set-cooked gel were buried.
33
Table 4. Estimated proportions of the buried and the
exposed tyrosine residues
Sample '850/830 N exposed N buried
Raw 1.18 0.91 0.09
Salted 1.14 0.85 0.15
Set 0.59 0.12 0.88
Cooked 3.1 3.47 -2.47
Set-cooked 0.22 -0.37 1.37
34
2.3.2.2 Secondary structure estimation from the amide I and ill
bands
The -CO-NH- amide or peptide bond has several distinct vibrational modes,
with the amide I band near 1650 cm" and the amide III band near 1250 cm'
1 1
being most easily identified in the Raman spectra of proteins. The exact
location of these bands depends on the secondary structure of the
polypeptide chain, and these features are therefore useful for estimating
secondary structure fractions of proteins. Due to possible overlap of bands
from the solvent water in the amide I region and from miscellaneous C-H
bending and aromatic ring vibrations in the amide III region, it is
recommended that both regions be analyzed in order to obtain a more reliable
interpretation of the changes in the secondary structure of proteins. The
Raman spectra of the surimi and gelled samples were, therefore, measured in
both regions, and maximum-likelihood deconvolution was used to enhance
the resolution of individual components contributing to the broad bands in
each of these regions.
The amide I band arises primarily from in-plane peptide C=0 stretching
vibrations and also has contributions from in-plane N-H bending vibrations.
Generally, proteins with high a-helical content show an amide I band centered
around 1645-1657 cm" while those with predominantly p-sheet structures

1
35
have a strong band at 1665-1680 cm" . Proteins with a high proportion of

1
undefined or random coil structure have the amide I band centered near 1660
cm' . The original and the deconvoluted Raman spectra in this region are
1
shown in Figures 9a and 9b, respectively. For the raw surimi, a major band at
1656 cm" suggestive of a-helical structure and a minor band near 1679 cm"
1 1
typical of 3-sheet structure are present. The salted surimi indicates a trend
towards uncoiling of helices with the major band centered near 1661 cm' , 1
while the minor band near 1683 cm" is suggestive of p-sheet or turns. For
1
each of the three remaining samples, increasing dominance of the band
assigned to p-sheet structure was evident.
The results of quantitative estimation of the secondary structure fractions
using the RSAP program for least squares analysis of the amide I region are
shown in Table 5. This algorithm has been reported to overestimate
antiparallel p-sheet content and underestimate a-helical content, but is useful
to assess relative changes in the secondary structure of proteins as a function
of different treatments (Przybycien and Bailey, 1989). The results suggest
that raw surimi contained predominantly a-helical structure with a smaller
proportion of antiparallel p-sheet, in agreement with visual inspection of the
deconvoluted spectra. Grinding with salt resulted in a slight increase in the
random coil or unordered fraction, at the expense primarily of the a-helical
fraction. After setting, similar proportions of a-helical and antiparallel p-sheet
36
Figure 9. Raman spectra of surimi and gels in the 1590-1720 cm" region
1
which includes the amide I band, a) Original spectra, b) Deconvolved

spectra'
37
Table 5. Secondary structure fractions estimated from the amide I band by
RSAP for the five types of samples
Treatment total a-helix antiparallel parallel p- total random
p-sheet sheet coil
Raw surimi 0.44 0.24 0.00 0.32
Grinding with salt 0.37 0.26 0.00 0.37
Setting 0.36 0.41 0.00 0.23
Cooking 0.09 0.31 0.20 0.40
Setting & cooking 0.20 0.49 0.00 0.31
38
were observed. Further increase in antiparallel p-sheet resulted upon
cooking after setting, while the cooked gel without prior setting showed an
increase in parallel p-sheet structure and a dramatic decrease in a-helix
content. Similar trends were noted by examining the amide III band in the
Raman spectra of the different samples. The amide III band results primarily
from the C-N stretching and N-H in-plane bending vibrations of the peptide
bond. It is less intense and located in a broad region from 1260-1300 cm" for1
proteins with high contents of a-helical structures, p-sheet structures usually
lead to a more intense band near 1238-1245 cm" while random coil structures
1
appear near 1250 cm" . 1

Figures 10a and 10b show the original and the
deconvoluted Raman spectra in the region which includes the amide III bands
from 1230-1300 cm" . Detailed inspection of the bands suggests that raw and
1
salted samples contain several bands in the amide III region typical of high a-
helical contents. The heat-set sample has increased the intensity of the
component bands near 1234-1242 cm' suggesting an increased proportion of

1
p-sheet structures. The trend towards higher p-sheet structure at the expense
of the a-helical content is even more evident in the cooked and set-cooked
gels. The cooked gel shows two main component bands in this region, while
the set-cooked gel shows only one major band at 1247 cm" with smaller
1
bands near 1280-1290 cm" . 1
39
1
which includes the amide III band, a) Original spectra, b) Deconvoluted

spectra
40
The Raman spectra in the amjde I and III regions thus collectively indicate a
trend toward decreasing a-helical content and increasing p-sheet content
upon setting, which became more pronounced after the cooking treatments.
These changes could be due to an unfolding of helical structures, followed by
the formation of sheet structures possibly through intermolecular interactions
between exposed hydrophobic residues. Chan et al. (1992) also reported an
unraveling of the coiled helical structure of fish myosin after heating to 50°C,
followed by exposure of hydrophobic residues. Moreover, Niwa (1992)
reported an increase in the hydrophobicity of actomyosin solutions caused by
setting. He also indicated that during cooking (kamaboko stage), the
interactions between the exposed hydrophobic sites become stronger.
2.3.3 C-H Stretching vibrations in the 2500-3400 cm' region

1
Figure 11 shows the Raman spectra of the five samples in this region, which
corresponds to various C-H stretching vibrations of primarily aliphatic
residues. For each sample, a major peak is shown between 2932 and 2939
cm" . A slight increase in wavenumber of the Raman shift was observed in the
1
gelled samples compared to the raw and salted surimi. However, information
on interpretation of this shift is lacking in the literature. Arteaga (1994)
reported that a slight shift to higher wavenumbers was observed upon
addition of water or deuterium oxide to simple organic solvents such as
41
Raw
Salted
-.-<
W Set
<D
a
i—i
(D
>
-<-( Cooked
-•->
cd
«—i
CD
I 1 I 1 1
3250 3000 2750 2500
Wavenumber ( c m - 1
)
1
showing the C-H stretching band
42
alcohols, dioxane and dimethyl sulfoxide. Several proteins also showed a
shift to higher wavenumbers in the presence of urea compared to non-
denaturing aqueous buffers. It was speculated that protein unfolding with
resulting exposure of methyl and methylene groups should produce spectral
changes to higher Raman shifts.
The normalized intensity as well as the area of the peak assigned to C-H
stretching vibrations decreased in the gelled samples compared to the raw
and salted surimi (Figure 12). Again, the interpretation of these results is not
clear, due to the scarcity of information in the literature on the study of amino
acid C-H stretching vibrations. One may speculate that the higher Raman
shifts and decrease in the intensity of the vibrations of the C-H stretching
band demonstrate changes in the environment of aliphatic C-H groups caused
by surimi gelation. Larsoon and Rand (1973) also found that the intensity of
the 2930 cm' peak increased with increased degree of polar environment of
1
the hydrocarbon chains.
43
Raw Salt Set Cook Set & cook
Treatment
Figure 12. Variation with treatment of the intensity and of the area of the
~2935 cm' peak
1
44
2.4 Conclusions
This study demonstrates the application of Raman spectroscopy to investigate
changes in the protein structure of intact surimi and gels from Pacific whiting.
Several important changes were observed during surimi processing. The
intensity of the peak assigned to disulfide bond S-S stretching vibrations
increased slightly after the setting treatment and considerably after the
cooking treatment. Setting decreased a-helical content and increased p-
sheet content. This trend was more pronounced after the cooking treatments.
A slight increase in the Raman shift and a decrease in intensity of C-H
stretching vibration of aliphatic residues was observed following the cooking
operations. These changes could be caused by an unfolding of helical
structures followed by interactions between the exposed hydrophobic
residues through formation of intermolecular sheet structures.
The gel properties, gel strength and fold score of the set and cooked gel were
higher than those of the cooked gel. The properties of the latter gel were
better than those of the gel that was subjected to the setting treatment alone.
Therefore, setting followed by cooking is the recommended method for the
manufacture of surimi gels.
45
Chapter 3. Optimization of processing conditions
Chapter 3
Optimization of the Setting and Cooking Conditions
for Kamaboko Making from Pacific Whiting Surimi
3.1 Introduction
One of the most important functional properties of surimi is its gelation property.
When surimi is chopped with salt (usually between 2 and 3% of surimi paste
weight), its myofibrillar proteins become soluble in water (Lee, 1984). When salt
is introduced, the salt ions bind to the oppositely charged groups exposed on the
protein surface and the proteins dissolve because of their increased affinity for
water (Niwa, 1992). When salted surimi is heated, it forms a fish gel structure
called kamaboko. The formation of a three dimensional gel network requires the
occurrence of at least three cross-links on every molecule (Niwa, 1992). Usually
slow setting at temperatures below 50°C precedes cooking at higher
temperatures. This results in a stronger gel than cooking without a slow set
(Lee, 1984).
46
Individual effects of heating parameters on surimi gelation have been studied.
For instance, Douglas-Schwartz and Lee ( 1988) tested the effect of setting for
20 minutes at four temperatures; 40, 60, 80 and 90°C. They found that setting of
Alaska pollock surimi and red hake surimi at 40°C resulted in gels with the best
cohesiveness and water holding capacity. For testing the quality of surimi gels,
Lanier (1992) suggested varying setting time and temperature but cooking
temperature was constant (90°C for 15 minutes). Still, there is a lack of
information about the combined effects of setting time and temperature as well
as cooking time and temperature on gel quality especially for surimi made from
Pacific whiting, an abundant species on the Pacific coast of North America. To
obtain this information, an experiment was designed to study the individual and
combined effects of setting and cooking conditions on the gel quality variables.
" Then optimization was performed to determine the processing conditions which
resulted in the best surimi gel.
What follows is a brief description of some the optimization techniques that were
used in this study.
47
C h a p t e r 3. O p t i m i z a t i o n o f p r o c e s s i n g c o n d i t i o n s
3.1.1 The Nelder-Mead simplex optimization method (Khuri and Cornell,
1987; Walters etal., 1991).
This is a 'direct search method" For k independent variables, it uses a
geometric figure; a simplex that is composed of k+1 points or vertices Pi
(i=1,2,..,k+1). The objective function Yj = f(Pj) is evaluated at each vertex. For a
minimization problem, the point having the maximum Y, P , is replaced by a new H
point according to one of these operations:
Reflection
The reflection of P is P* which is defined by:

H
P* = ( 1 + a ) C - a P H
where C is the centroid of the points other than P H , a > 0 is the reflection
coefficient (i.e., a = 1/2, 2/3, or 1). If Y < Y* = f(P*) < Y , [Y = min (Yi), 1 < i <
L H L
k+1], then P is replaced by P* and the process is restarted.

H
Expansion
If Y* < Y , then P* is expanded to P by using the following equation:

L E
48
P E =YP* + ( 1 ^ ) C
where y > 1 is the expansion coefficient (i.e., y = 1.5). If Y E = f(P ) < Y , then
E L
replace P by P and restart. Otherwise, if Y = f(P ) > Y , then replace P by P*

H E E E L H
and restart the process.
Contraction
If the reflection of P to P* results in Y* > Y| for all i * H, then a new P is to be

H H
defined as the old P or P* selecting the point that has the lower Y value. Let us
H
define P as:
c
P C = P P H + (1-P)C
where p < 1. If Y = f(Pc) > min (Y ,Y*) then replace all the values of Pj with
c H
(Pi + P )/2 and restart the process. Otherwise, replace P with P and restart.
L H C
To check the optimization results obtained by the Nelder-Mead simplex method,
another type of optimization technique was used. This method is a gradient
based method that is described below.
49
3.1.2 The Gradient based optimization methods
These optimization methods use the partial derivatives of the objective function
in order to find the direction of search. For a minimization problem and when the
steepest descent method (a common gradient based method) is employed, the
gradient vector at a point gives the direction of the greatest decrease in the
objective function and, therefore, guides the transition from one point to another
until an optimal value is found (Edgar and Himmelblau, 1988). The NLPQLO
gradient based method uses a quadratic approximation of the objective function
(Vaessen, 1984).
The simplex method and the gradient based method are local optimization
methods, e.g., they are only able to find a single minimum even though other
minima having lower objective function values may also be located in the
constrained search field. A global optimization method was used to assess the
results obtained by the two local techniques. This global method is the Level
Set Programming technique.
3.1.3 The Level Set Programming (LSP) method
The fact that LSP is a global optimization technique is very useful when dealing
with problems having many local optima. The LSP theory was initially presented
50
C h a p t e r 3. O p t i m i z a t i o n o f processing c o n d i t i o n s
by Chew and Zheng (1988). This nonlinear programming technique belongs to
the direct search category. It does not need gradient evaluations and is mainly
based on the determinations of the objective and constraint (if any) functions at
solution points. An optimization software based on this method was developed
by Yassien (1993). The method consists of randomly generating solutions
having objective functions less than or equal to a pre-selected value called the
level set value (LSV). The mean of these values defines a new LSV and hence
a new level set interval. The search process continues at progressively
decreasing LSV until the convergence criteria are satisfied. Then, the global
solution or set of solutions is determined (Yassien, 1993).
The objective of this study was to determine the optimal setting time and
temperature and cooking time and temperature resulting in Pacific whiting surimi
gel with the best combination of gel strength, fold score, color and expressible
liquid.
The Response surface methodology (RSM), central composite design was used
in order to correlate the response variables to the independent variables. Table
6 depicts the experimental design. For four independent variables, it consists of
51
Table 6. Experimental design for the optimization of surimi heating conditions
Factors CODES
-2 -1 0 1 2
X, setting temperature (°C) 20 30 40 50 60
X2 setting time (min.) 0 10 20 30 40
X3 cooking temperature (°C) 70 75 80 85 90
X4 cooking time (min.) 5 15 25 35 45
X1 X2 X3 X4
0 0 0 0
0 0 0 0
-2 0 0 0
2 0 0 0
0 -2 0 0
0 2 0 0
0 0 -2 0
0 0 2 0
0 0 0 -2
0 0 0 2
0 0 0 0
0 0 0 0
52
2 (16) factorial settings, 2x4 (8) axial settings and 6 center point replicates
4
(Khuri and Cornell, 1987). The four independent variables were setting
temperature (X^, setting time (X ), cooking temperature (X3) and cooking time
2
(X4). Experiments were carried out on Pacific whiting surimi donated by Ucluelet
Seafood Processors Ltd. (Ucluelet, B. C , Canada). For each set of processing
conditions, surimi was mixed with 3% salt (total weight basis) before being set
and cooked as in section 4.3.2.2. The setting and cooking conditions are
depicted in Table 6. The quality of the obtained surimi gels was tested by
performing the following quality tests.
3.2.1 Gel strength (compression test)
This test is an indication of gel cohesiveness (Lee et al., 1987). The procedure
for this test can be found in section 2.2.3.
3.2.2 Fold score
This test indicates gel elasticity (Lee, 1984). It was performed according to the
procedures outlined in section 2.2.3.
53
C h a p t e r 3. O p t i m i z a t i o n o f processing conditions
3.2.3 Gel color test
The whiteness of a surimi gel sample (thickness = 50 mm, diameter = 30 mm)
was measured using a color measurement instrument (Hunterlab, Model
Labscan II, Reston, Virginia). The test consists of measuring L (lightness) on a
0-100 scale from black to white in shades of gray, a denoting redness (if
positive) or greenness (if negative), and b indicating yellowness (if positive) or
blueness (if negative). The "whiteness" index, C, for the overall color evaluation
is (Lanier, 1992):
C = 100-[(100-L) +a + b ] 2 2 2 05
3.2.4 Expressible liquid measurement
Expressible liquid is a measure of the water binding ability of the gel. It was
measured by compressing a surimi gel cylinder (20 mm height and 30 mm
diameter), using the Instron Universal Instrument (Model 1122, Canton,
Massachusetts) at a cross head speed of 10 mm / min, into a 1 mm thick disk in
1.9 minutes and absorbing the expressed moisture in two pieces of filter paper
for five minutes. Free water is more easily expressed upon compression than
bound water. Expressible liquid, Ei, is defined as (Niwa, 1992):
54
E, (%)=(Wi-W )x 100/ Wi f
where Wj is the weight of the surimi gel before compression while W is the f
surimi gel weight after compression.
These quality tests were conducted in triplicates.
The optimization techniques used were the Nelder-Mead simplex method
(Arteaga, 1992), the NLPQLO gradient based method (Vaessen, 1984) and the
Level Set Programming method (Yassien, 1993). Xi had a lower limit of 22°C
and an upper limit of 45°C. X and X 4 varied between 10 and 40 minutes. The
2
lower and upper limits for X 3 were 70 and 90°C, respectively.
Table 7 summarizes the results of the optimization experiments. For each set of
independent variables, setting temperature (X^, setting time (X ), cooking 2
temperature (X3) and cooking time (X4), it shows the corresponding response
variables, namely gel strength (G ), fold score (F ), color (C) and expressible
s s
liquid (Ei). G values were higher for cooking temperatures of 80°C and above.
s
The fold score values were also better at these cooking temperatures. A lower
55
Table 7. Experimental results

j
Row X, x 2 Xa X4 Gs 1 c 4
E,
(°C) (min.) (°C) (min.) (N.mm) Fs (%)
1 30 10 75 15 130 2.0 55.7 18.1
2 50 10 75 35 940 3.5 55.1 16.0
3 30 30 75 35 230 3.0 56.4 16.7
4 50 30 75 15 330 2.5 55.6 16.2
5 30 10 85 35 2860 5.0 55.8 19.1
6 50 10 85 15 3080 4.5 55.6 13.0
7 30 30 85 15 3950 6.0 57.6 19.3
8 50 30 85 35 720 3.0 55.2 14.5
9 40 20 80 25 2540 5.0 55.9 19.5
10 40 20 80 25 2160 5.0 56.3 19.5
11 30 10 75 35 520 4.0 56.5 16.5
12 50 10 75 15 150 2.5 54.1 17.4
13 30 30 75 15 110 1.5 56.9 18.9
14 50 30 75 35 720 3.0 56.8 14.4
15 30 10 85 15 3680 5.0 56.8 21.1
16 50 10 85 35 1470 3.5 55.6 14.9
17 30 30 85 35 3640 3.5 55.5 20.3
18 50 30 85 15 1270 3.0 55.4 14.3
19 40 20 80 25 2120 5.0 54.9 18.8
20 40 20 80 25 2940 6.0 59.1 12.4
21 20 20 80 25 3050 6.0 68.1 13.2
22 60 20 80 25 3050 6.0 65.4 12.7
23 40 0 80 25 2920 4.0 59.6 13.1
24 40 40 80 25 2800 6.0 59.9 12.1
25 40 20 70 25 340 2.0 67.2 16.7
26 40 20 90 25 1620 3.0 67.0 14.8
27 40 20 80 5 2810 6.0 58.3 10.4
28 40 20 80 45 2150 5.0 53.5 19.1
29 40 20 80 25 2960 6.0 60.7 12.5
30 40 20 80 25 2960 6.0 61.4 13.0
'Arithmetic mean (n=3)

Chapter 3 .Optimization of processing conditions
fold score, however, was observed for surimi cooked at 80°C without previous
setting. This shows the effect of setting on the gel elasticity and supports the
findings of Lanier et al. (1982) who reported that surimi gels prepared by setting
prior to cooking were more elastic than those that were directly cooked. Setting
at 50°C for 30 minutes followed by cooking at 85°C for 35 minutes resulted in
weaker and less elastic gels than were obtained under the same cooking
conditions but at a setting temperature of 30°C for 30 minutes. This could be
due to the gel softening phenomenon, called modori in Japanese, which usually
occurs when surimi is heated at temperatures between 50 and 70°C. Niwa
explained that proteolytic degradation of myosin at the modori temperature
range may be the cause of gel softening. Most of the C values were higher than
55 which corresponds to grade 4 according to the classification reported by
Lanier (1992). (Grade 1 corresponds to C > 60.0.) Seven out of eight of the
combinations that resulted in a fold score of 6 also resulted in low expressible
moisture. This supports the deduction of Tagagi (1973) that the more elastic the
surimi gel the lower the expressible moisture. The six center point replicates
were reasonably reproducible for G , F and C. However, three points resulted

s s
in high E, whereas the other points gave low E, values. Since these six points
resulted in high fold scores, it is thought that the high values of E observed for
t
three points could be due to experimental error. In most of the cases, less
moisture was expressed from gels that were cooked at 80°C and above.
57
Each one of these response variables was fitted by an empirical equation in
order to correlate them to the independent variables. The empirical equation
has the general form:
Y = Bo + £/3iXi + f BnXi j
2
+2 ^PaXiXj
i=l i=l i=l ;'=i+l
where X 4 are the input variables which influence the response variable Y
while Po, Pi(i=1,...,4) and Py (i=1,...,3; j=i+1,..., 4) are the model parameters. This
is the second order model (SOM) used in the Response Surface Methodology
(Khuri and Cornell, 1987).
3.3.1 Curve fitting results
Curve fitting was carried out using the Systat 1990 (Systat, Inc., Evanston, IL)
software. The parameter estimates obtained by the multiple, stepwise
regression of the measured gel strengths are shown in Table 8. Only the terms
that significantly affected the model (P < 0.05) are included in the table and were
taken into account in the optimization procedures which are discussed in section
3.3.2. The logarithm of the gel strength was used in the regression analysis as
the unaltered values, presumably because of their two-orders-of-magnitude
range (from 110 to 3950 N.mm), yielded poor regression results (R < 0.50). 2
Table 8 shows that cooking temperature had the strongest influence on gel
58
Table 8. Regression results where "log (G )" is the response variable

s
Coefficient R z
Standard Error of
Estimate
Po -114.024 0.858 0.180
01 0.284
p 3
2.606
P4 0.285
Pl3 -0.004
P33 -0.014
P34 -0.004
59
strength while setting temperature and cooking time had less effect on G . s
However, setting time, at least in the range of 10 to 40 minutes, did not affect G . s
Table 9, which compares the experimental and estimated values of G , F , C and s s
Ei, also demonstrates that the G s data were well fitted by the SOM. The
experimental data of F were also reasonably well fitted by the SOM (Tables 9
s
and 10). F was mainly dependent on the cooking temperature. The three other
s
independent variables affected F only slightly. Table 11 shows that the curve
s
fitting results for the color variable were worse than those of the other response
variables. The higher the setting temperature and/or the cooking temperature,
the lower the whiteness index of the surimi gel. Setting time and cooking time
did not affect color. Measured E, data were well fitted by the SOM (Tables 9 and
12). The model parameters listed in Table 12 demonstrate that the higher the
cooking temperature and/or time, the lower the expressible liquid. For all four
response variables, the quadratic parameters had very little influence on the
overall fit.
The fact that cooking temperature was the variable that had the most influence
on the gel cohesiveness, elasticity and water holding capacity shows the
importance of the cooking stage on the gel texture. At the cooking stage, as
found in Chapter 2, more disulfide interactions take place and the p-sheet
content increases. This could result in better gel texture.
60
Table 9. Experimental and estimated values of the 4 response variables
Row# log G s log G s Fs Fs

C C E, E,
(exp.) (est.) (exp.) (est.) (exp.) (est.) (exp.) (est.)
1 2.12 2.11 2.00 1.96 55.7 56.8 18.1 19.2
2 2.97 2.88 3.50 2.76 55.1 54.9 16.0 15.2
3 2.37 2.55 3.00 1.61 56.4 55.5 16.7 17.3
4 2.52 2.44 2.50 1.68 55.6 56.2 16.2 17.1
5 3.46 3.52 5.00 3.53 55.8 55.4 19.1 19.7
6 3.49 3.39 4.50 4.17 55.6 68.7 13.0 14.4
7 3.60 3.78 6.00 4.64 57.6 56.7 19.3 19.6
8 2.86 3.14 3.00 1.61 55.2 67.4 14.5 14.6
9 3.40 3.33 5.00 4.61 55.9 58.3 19.5 13.0
10 3.33 3.33 5.00 4.61 56.3 58.3 19.5 12.0
11 2.72 2.55 4.00 2.33 56.5 55.5 16.5 17.3
12 2.17 2.44 2.50 2.40 54.1 56.2 17.4 17.1
13 2.02 2.11 1.50 1.25 56.9 56.8 18.9 19.2
14 2.86 2.88 3.00 2.05 56.8 54.9 14.4 15.2
15 3.57 3.78 5.00 5.36 56.8 56.7 21.1 19.6
16 3.17 3.14 3.50 2.32 55.6 67.4 14.9 14.6
17 3.56 3.52 3.50 2.80 55.5 55.4 20.3 19.7
18 3.10 3.39 3.00 3.45 55.4 68.7 14.3 14.4
19 3.33 3.33 5.00 4.61 54.9 58.3 18.8 13.0
20 3.47 3.33 6.00 4.61 59.1 58.3 12.4 13.0
21 3.48 3.36 6.00 4.99 68.1 52.6 13.2 21.9
22 3.48 3.30 6.00 4.23 65.4 64.0 12.7 14.7
23 3.47 3.33 4.00 2.54 59.6 58.3 13.1 13.0
24 3.45 3.33 6.00 1.10 59.9 58.3 12.1 13.0
25 2.53 0.95 2.00 -1.54 67.2 55.1 16.7 16.5
26 3.21 2.87 3.00 1.42 67.0 67.5 14.8 16.3
27 3.45 3.24 6.00 5.35 58.3 59.1 10.4 21.8
28 3.33 3.43 5.00 3.87 53.5 56.5 19.1 20.0
29 3.47 3.33 6.00 4.61 60.7 58.3 12.5 13.0
30 3.47 3.33 6.00 4.61 61.4 58.3 13.0 13.00
61
Table 10. Regression results where "F " is the response variable
s
Coefficient R 2
Standard Error of
Estimate
Po -354.198 0.80 0.627
Pi 0.631
p 2
0.243
p 3
8.211
P4 0.848
Pl3 -0.008
P22 -0.007
P33 -0.047
P34 -0.011
62
Table 11. Regression results where "C" is the response variable
Coefficient R 2
Standard Error of
Estimate
Po 388.221 0.667 1.911
Pi -4.776
p 3
-6.602
Pl3 0.063
P33 0.029
P44 -0.001
63
Table 12. Regression results where "E" is the response variable
Coefficient R 2
Standard Error of
Estimate
Po 245 0.892 0.895
P3 -5.1
P4 -1.868
P11 0.013
Pl3 -0.016
P33 0.034
P34 0.010
P44 0.020
64
The generated equations for the response variables were then used to
determine the optimal processing conditions.
3.3.2 Optimization results
Using the empirical equations found for each variable as objective functions,
three optimization techniques were used to determine the best set of
setting/cooking times and temperatures in each case. These were the Nelder-
Mead Simplex method, the Level Set Programming method, and the NLPQLO
Gradient method.
3.3.2.1 Results of Nelder-Mead Simplex optimization
The results of optimizing one response variable at a time (maximizing G , F and

s s
C, and minimizing Ei) using the Simplex method are shown in Table 13. This
table contains the optimal processing conditions (Xj, i=1,2 4) along with the
corresponding values for the four response variables. The optimized values of
log (G ), F
s S| C and E, were 4.08, 6.41, 70.8 and 12.3, respectively. Apart from
the Ei value, the other optimized values were better than the best experimental
results. The processing conditions corresponding to a maximized C, 70.8,
resulted in a negative F value. This is a further indication that SOM fit of the
s
color values was relatively poor.
65
Table 13. Optimization of individual response variables using the Nelder-Mead
Simplex method
Variable Maximize G s
Maximize F s Maximize C Minimize Ei
* (°C) 22.0 24.1 45.0 45.0
X (min.)
2
36.1 21.1 40.0 24.3
XaCC) 87.4 85.9 90.0 81.1
X 4 (min.) 10.0 10.0 40.0 25.8
log (G ) s
4.08 4.00 2.23 3.40
F s
3.98 6.41 -4.80 4.26
C 51.7 53.2 70.8 60.8
E, (%) 28.3 26.0 20.3 12.3
66
3.3.2.2 Results of NLPQLO Gradient optimization
Table 14 depicts the results of optimizing the response variables using the
NLPQLO gradient method. These results include the optimal processing
conditions and the corresponding response variable values. The optimized
values of log (G ), F , C and Ei were 4.08, 6.64, 72.7 and 12.3, respectively. The
s s
maximized F and C values were higher than those obtained by the Nelder-Mead
s
Simplex method indicating that the Simplex optimizer likely found only local
maxima for these two response variables. In addition, the processing conditions
corresponding to a maximized C resulted in better values of log (G ), F

s s and Ei
than those obtained by the Simplex technique.
3.3.2.3 Results of LSP optimization
The optimization results obtained using this method were almost the same as
those obtained by the NLPQLO gradient method (see Table 15). The optimal
setting time values for G , C and Ei were considerably different from those
s
obtained by the gradient method. However, these response variables, as
indicated in Tables 8, 11 and 12, respectively, were independent of setting time
over the range from 10 to 40 minutes. Hence, the results of the global LSP
method confirmed the results of the local gradient method for the independent
variables limits specified in section 3.2.
67
Table 14. Optimization of individual response variables using the NLPQLO
Gradient method
Variable Maximize G s Maximize F s Maximize C Minimize Ei
* (°C) 22.0 22.0 45.0 45.0
X (min.)
2 20.0 20.0 20.0 20.0
XaCC) 87.5 84.9 90.0 81.2
X 4 (min.) 10.0 10.0 10.0 25.8
log (G ) s
4.08 3.99 3.14 3.40
F s
6.34 6.64 3.13 4.53
C 51.7 52.1 72.7 60.9
E, (%) 28.4 27.0 18.5 12.3
68
Table 15. Optimization of individual response variables using the LSP method
Variable Maximize G s Maximize F s Maximize C Minimize E t
X, (°C) 22.0 22.0 45.0 45.0
X (min.)
2 23.8 17.4 30.8 28.0
X3(°C) 87.5 85.0 90.0 81.2
X 4 (min.) 10.0 10.0 10.0 25.8
log (G ) s 4.08 3.99 3.14 3.40
F s
6.09 6.68 1.94 3.80
C 51.7 52.0 72.7 60.9
E. (%) 28.4 27.0 18.5 12.3
69
3.3.2.4 Multiobjective optimization
As can be seen in Tables 13 to 15, the optimal conditions for the optimization of
one response variable (e.g., G ) did not give satisfactory results for the other
s
response variables (i.e., F , C and Ei).

s Therefore, multiobjective optimization
had to be performed in order to simultaneously optimize the four response
variables in question. To fulfill this task, the multiobjective problem was
transformed into a single objective problem. This was achieved by dividing each
variable by its maximum experimental value and forming a single objective
function, Z, as the weighted sum of these normalized variables (de Neufville,
1990). Each variable was normalized in this fashion in order to put all the
response variables on the same scale. The expression for Z is:
Z = w, [log(G )/3.60]+ w (Fs/6)+ w (C/68.1) - w (E,/21.1)

s 2 3 4
where Wi,...,w are weights that are chosen arbitrarily taking into consideration
4
the importance of each response variable so as to generate acceptable results
for all the four response variables. The terms involving gel strength, fold score
and color were added because they were to be maximized. However, the
expressible liquid term was subtracted because it was to be minimized (Yassien
(1993).
70
Several combinations of w (k=1 k 4) were tried. Some of these combinations
along with the corresponding results are shown in Table 16. Those results were
obtained using the LSP method. Emphasis was put on maximizing gel strength
and fold score without excessively compromising color or the expressible liquid.
In most cases, the color variable was assigned a low weight, 0.1, because its
objective function was not well fitted to the measured color data. The Z values
could not be compared because the weights in each row were different. The row
in the bold font seems to give satisfactory results for all four response variables.
These results consisted of a gel strength of 3980 N.mm, a fold score of 5.4, a gel
whiteness index of 60.5 and an expressible liquid of 15.6%, corresponding to
setting at 39.2°C for 17.4 minutes followed by cooking at 82.8°C for 14.7
minutes.
71
Table 16. Multiobjective optimization results
w 2 W 3 w 4 Xi x 2 Xa X4 log G s F s
c E, z
1 1 1 1 44.8 17.0 83.2 16.9 3.5 5.0 62.4 13.9 2.0
1 6 0.1 3 36.6 17.4 83.5 11.3 3.6 5.8 59.7 18.2 4.4
1 5 0.1 3 39.2 17.4 82.8 14.7 3.6 5.4 60.5 15.6 3.4
1 6 0.1 3.5 38.5 17.8 83.0 14.1 3.6 5.5 60.3 16.1 4.0
1 6 0.1 4 40.9 17.4 82.6 16.8 3.5 5.2 61.0 14.5 3.6
1 8 0.1 5 40.3 17.4 82.6 15.9 3.5 5.3 60.7 14.9 4.7
1
9 0.1 5 38.9 17.5 83.1 14.1 3.6 5.5 60.6 16.0 5.6
72
3.4 Conclusions and recommendations
The experimental results on the effects of processing conditions on surimi gel
quality showed that high cooking temperatures (80°C and above) resulted in
better gel cohesiveness, elasticity and water holding capacity. They also
showed the importance of the setting treatment prior to sample cooking.
However, setting should be performed at temperatures below 50°C. The data
were well fitted by a second order model which allowed four objective functions
to be generated for the four response variables; gel strength, fold score, color
and expressible liquid. The optimal processing conditions corresponding to
individually maximizing G , F and C, and minimizing Ei were determined using

s s
the Nelder-Mead Simplex optimization and the NLPQLO gradient optimization.
The results obtained using the latter method appeared to be closer to the global
optima than those of the Simplex method. The results obtained using a global
optimization method, LSP, essentially confirmed the results of the gradient
method. Multiobjective optimization was also conducted to simultaneously
optimize the four response variables. It gave a set of setting and cooking times
and temperatures which led to satisfactory values for all four response variables.
However, these optimal processing conditions are only applicable to Pacific
whiting kamaboko sausages having the size used in this project. The same
73
methodology can be followed to determine the optimal processing conditions for
other species and for other surimi-based products.
The ingredients used for this work were Pacific whiting surimi and salt. For
further studies, it is recommended that the gel texture improving ingredients
(GTII) such as starch, egg white or whey protein concentrate be taken into
account in the RSM experimental design. The concentrations of these
ingredients would therefore be considered as independent variables to be
optimized. This will allow a comprehensive study of the effects of setting and
cooking temperatures and times along with the effects of the GTII on the surimi
gel characteristics.
74
Chapter 4. Production of salmon and herring kamaboko
Chapter 4
Kamaboko Production from Roe Herring
and Pink Salmon
4.1 Introduction
Alaska pollock has been the most popular species for the production of
surimi. This is due to its accessibility, subtle flavor and relatively large size,
among other characteristics (Holmes et al. 1992). However, with the increase
in surimi production, the stocks of Alaska pollock have decreased
dramatically. For instance, the pollock biomass in the eastern Bering Sea
was estimated at 5.3 million metric tons in 1989, a 47% decline since 1986
(Holmes et al., 1992). This situation has encouraged the use of other species
such as whiting, cod and New Zealand hoki.
Roe herring (Clupea pallasi) is a species that is abundant on the coast of
British Columbia, Canada. Its annual harvest is between 15,000 and 40,000
tons. The catch is almost exclusively aimed at collecting herring roe, a very
75
popular commodity in Japan. The male fish and the remainder of the female
fish, after roe collection, are either wasted or reduced to fish meal. Making
surimi or a related commercial product from the leftover herring could be a
good alternative. Kamaboko of average quality was obtained from fresh North
Sea herring (Hastings et al., 1990). The quality of kamaboko was worse
when North Sea herring was frozen at -30°C for six months. Nakai (1993)
found that no kamaboko could be made from herring surimi that was frozen at
-35°C for three months.
In this study, an attempt was made to produce acceptable surimi-based
products from herring that was frozen at different conditions in the form of raw
fish. However, due to the limited availability of roe herring and because its
small size makes surimi production difficult, another species was also tried.
Pink salmon (Oncorynchus gorbuscha) is very abundant on the British
Columbian coast and is mainly used for canning. It is characterized by its
large size and relatively low cost. Therefore, Pink salmon was also utilized to
produce surimi and kamaboko.
When washed fish mince is the only component of a surimi-based product,
the texture tends to be rubbery and freeze-thaw stability is often poor (Lee et
al., 1992). To improve the texture of the final product, several ingredients can
be added. In this study, the ingredients used were salt, whey protein
76
C h a p t e r 4. P r o d u c t i o n o f s a l m o n and herring k a m a b o k o
concentrate and wheat starch. Salt was used to improve the solubility of
surimi myofibrillar proteins. Kim and Lee (1987) explained that starch is
mainly used as a filler ingredient. Upon heating, it swells and increases water
uptake. This improves the firmness and the cohesiveness of the surimi gel
matrix. Wu et al. (1985) reported that starch does not directly interact with the
surimi protein matrix nor significantly influence its formation. Several studies
have reported the use of whey protein concentrate in the manufacture of
surimi-based products (Burgarella et al., 1985; Chung, 1990; Lee and Kim,
1986). It is also used as a filler. Its gel strengthening effect has been
reported by Lee et al. (1992b). Thus, there was a need to investigate the
combined effects of salt, wheat starch and whey protein concentrate on
kamaboko made from pink salmon and roe herring.
4.2 Objectives
a. To study the effect of frozen storage conditions on the kamaboko
making ability of roe herring.
b. To optimize a formulation consisting of surimi, salt, whey protein
concentrate and wheat starch for kamaboko made from pink salmon surimi
and from roe herring.
77
4.3.1 Materials
Frozen, whole, roe herring was purchased from a local fishing company (Sung
Fish Company, Vancouver, B. C.) on May 17, 1994. The company kept this
fish frozen at -40°C for approximately one month (since mid-April 1994). It
was transported to our laboratory where 11 pounds (lb) were stored at -83°C,
70 lb at -45°C and 70 lb at -35°C. The fish comprising the last sample were
soaked in glycerol (10% solution) for 24 hours before further frozen storage.
The cryoprotective effect of glycerol has been reported by Matsumoto and
Noguchi (1992).
Due to the limited quantity of herring in our possession, pink salmon was
selected for the tests relating to the optimization of the kamaboko formulation.
Pink salmon is suitable for this type of study because the surimi preparation
from it is less labor intensive and because there is also a commercial
incentive to produce surimi from this species. It was therefore intended to
optimize the kamaboko formulation with salmon surimi and to use the
resulting optimum conditions to prepare roe herring kamaboko. Fresh pink
salmon was purchased at the end of July 1994. It was kept frozen at -35°C for
78
two months before salmon surimi was made from it. After four months of
storage at -35°C, the salmon surimi was then used in the optimization
experiments. To improve the texture of salmon kamaboko, several
ingredients were used. These are salt, modified whey protein concentrate
(WPC) (Alaco Surimi Plus), wheat starch (WS) and water. The ingredient
combinations and their respective concentrations are listed in Table 17.
Two Japanese surimi-based products (Maruu Kamaboko Shiro and Kibun
Hanpen-L), which were made from Alaska pollock, were used as commercial
target products.
4.3.2 General procedures
4.3.2.1 Surimi making
The fish was filleted, minced, washed 4 times in a 0.3% salt solution (the
volume of the solution was five times the volume of the fillets), and mixed with
cryoprotectants [4% sucrose (meat weight basis), 4% sorbitol, 0.15%
tripolyphosphate and 0.15% sodium pyrophosphate] in a silent cutter (Hobart
Manufacturing Co., Model 84142, Don Mills, Ontario) for 5 minutes. These
procedures were performed at 4°C. The surimi was stored frozen at -35°C.
79
4.3.2.2 Kamaboko making
Surimi, partially thawed at room temperature for one hour, was chopped, in a
vacuum cutter (Stephan, Model VCM-5, Columbus, Ohio), at low speed for
two minutes. Then 3% salt (total weight basis) was added and the mixture
was mixed at low speed for three minutes. Finally, mixing was performed
under vacuum (to prevent oxidation) at high speed for one minute. Circulating
ice chilled water in a jacket allowed the mixing to be carried out at
temperatures below 10°C as recommended by several researchers (i.e., Lee
et al., 1992a; Douglas-Schwartz and Lee, 1988; Lanier, 1992). When WS
and WPC were used, the mixing procedure was different. Surimi chopped at
low speed for two minutes, was mixed with salt at low speed for two additional
minutes. The paste was then blended with WS (premixed with water) and
WPC for two more minutes at low speed. The paste was finally mixed at high
speed and under vacuum for one minute. Using a meat stuffer (Kitchenaid,
Model K5-A, Troy, Ohio) the paste was stuffed into sausage casings having a
diameter of 30 mm and a length of 140 mm. These sausages were then
subjected to setting at 45°C for thirty minutes in a Blue M water bath (Model
MW-1120A-1, Blue Island, IL) and then cooking at 90°C for half an hour (in a
second Blue M water bath). The end product, kamaboko sausages, were
immediately cooled in ice for fifteen minutes and kept at room temperature
overnight before testing. The setting and cooking conditions were selected in
80
light of previous experiments aimed at optimizing the processing conditions
for kamaboko made from fresh herring (Nishimura et al., 1993).
4.3.2.3 Measurement of surimi gel properties
The quality of the kamaboko sausages was tested by performing the punch
and die test and the fold test. The punch test provides an indication of the gel
firmness, while the fold test measures the elasticity of the product (Lanier,
1991). The punch tests were performed using an Instron Universal Testing
Instrument (Model 1122, Canton, Massachusetts). A surimi gel cylinder,
having a diameter of 30 mm and a height of 20 mm, was centered under a
plunger (5 mm diameter). The plunger goes through the sample and then
through an orifice (8 mm diameter). The cross head speed was 10 mm / min.
The variation of the penetration force with time was recorded. The gel
strength is the product of the force and the distance at the point of rupture
(see Figure 4). The fold test was carried out according to the description
given in section 2.2.3.
4.3.3 Roe herring samples
On May 18,1994 surimi was made from about 30 pounds of whole herring
(one day after its purchase). The surimi was stored frozen at -35°C. On the
81
following day, kamaboko was made from this surimi and its quality was
evaluated by performing the punch and the fold tests. Surimi and kamaboko
were made from herring stored at -35°C and at -45°C for forty days. Fish
stored at the latter temperatures for 130 days was used to produce surimi and
kamaboko. Finally, surimi and kamaboko were made from frozen fish stored
at -35°C, at -45°C and at -83°C for 190 days.
4.4.1 Frozen storage experiments
After 40 days of storage at -35°C the glycerol soaked roe herring gave a
punch test score of 51 N.mm and a fold score of 3. The sample which was
stored at -45°C for the same period of time resulted in a punch test score of
61 N.mm and a fold score of 3. By comparison, kamaboko made from frozen
herring before the cold storage was initiated (i.e., at the time when the frozen
herring was purchased from Sung Fish Co.) gave a punch test score of 61
N.mm and a fold test value of 3. From these results it appears that the fish
stored at -45°C without the use of glycerol gave the same gel strength and
fold test values as the sample before prolonged frozen storage. These results
82
are similar to the findings of Fukuda (1991) in which he concluded that freeze
damage of surimi made from cod, salmon, sardine and mackerel and frozen
for six months could be reduced at cold storage temperatures lower than
-AO°C.
However, Figure 13 shows that after 130 days of storage, the sample stored
at -45°C had a gel strength of 43 N.mm and a fold score of 2 while the sample
soaked in glycerol and stored at -35°C had a gel strength of 37 N.mm and a
fold score of 2. Thus, a notable decrease of gel strength occurred between
40 and 130 days of frozen storage. After 190 days of frozen storage, the
sample stored at -83°C had a gel strength of 57 N.mm and a fold score of 3.
This sample had maintained reasonable gel quality. By comparison, the gel
strength values of the samples stored at -45°C and at -35°C for 190 days
decreased further to 36 and 33 N.mm, respectively. These two samples had
a fold score of 2. Hastings et al. (1990) also found that the frozen storage of
whole herring for six months at -30°C resulted in a kamaboko having a gel
strength about half of that of kamaboko made from fresh herring.
The deterioration of the gel making ability of roe herring after prolonged
frozen storage could be due to the denaturation of myofibrillar proteins, a
phenomenon suggested by many researchers (i.e., Watabe and Hashimito,
83
70 - x — 3 5 C + glycerol
—I 45 C
-83 C
20 4-
10 4-
50 100 150 200
Time [days]
Figure 13. Variation of roe herring kamaboko gel strength with frozen storage
84
1987; Matsumuoto and Noguchi, 1992). This denaturation seems to be
prevented by frozen storage at ~45°C up to forty days. Frozen storage at
-83°C for 190 days maintained the gel firmness and elasticity. These findings
are similar to those of Watabe and Hashimito (1987) who reported a decrease
in the myofibrillar protein fraction caUsed by frozen storage of Alaska pollock
surimi at -20°C and at -30°C. This decrease, however, did not occur for the
storage temperatures of -40 and -80°C. These researchers concluded that
frozen storage at -40°C or lower temperatures prevented protein denaturation
and maintained the gel making ability of Alaska pollock surimi.
Because of the complexity of the mechanisms responsible for the
deterioration of the gel making ability of frozen fish muscle, it is
recommended, for future studies, that Raman spectroscopy be used to
investigate the changes that take place during the freeze denaturation of fish
proteins.
4.4.2 Optimization experiments
The results of the formulation optimization experiments are shown in Table
17. Figure 14 shows the variation of gel strength with whey protein
concentrate and wheat starch. The combined effects of the latter ingredients
85
Table 17. Effects of different formulations on surimi gel texture
%WPC %WS % Water Gel Strength Fold % Moisture

# Starting Material added (N.mm) Score
1 SS S
0 0 0.00 58.9*+ 5.5" 3 71.8
2 SS 2 3 2.24 65.4±14.7 4 69.6
3 SS° 2 5 7.08 79.6+12.9 6 69.2
4 SS 2 7 11.92 68.3±9.6 6 70:2

5 SS 4 3 7.80 86.2±7.7 4 70.4
6 SS 4 5 12.64 61.7±7.3 6 69.6
7 SS 4 7 17.49 45.6+4.5 6 69.3
8 SS 6 3 13.36 53.6±7.7 4 69.8
9 SS 6 5 18.20 50.5±6.1 5 70.1
10 SS 6 7 23.00 51.3+6.6 6 69.3
11 SS 4 0 0.00 72.2+6.2 3 69.4
12 SS 0 5 0.00 64.7±5.4 4 69.3
13 SS 2 5 0.00 104.7±7.8 4 66.7
14 HS h
2 5 7.00 74.4* ±3.3 6 NM"
15 MKS m
99.7*+8.6 6 74.2
16 KH-L k
84.2* ±2.3 6 71.5
8
Salmon surimi plus 3 % salt
h
Herring surimi plus 3 % salt
0
Optimal formulation
m
Maruu Kamaboko Shiro
k
Kibun Hanpen-L (Boiledfishcake)
" Not measured
* Arithmetic mean (n=5-7)
* Arithmetic mean (n=3)
** Standard deviation
86
Figure 14. Combined effects of WPC and WS on salmon surimi gel strength
87
on the fold score are depicted in Figure 15. Formulation 5, consisting of
82.2% salmon surimi (total weight basis), 3% salt, 4% WPC, 3% WS and
7.8% water, gave a gel strength of 86.2 N.mm and a fold score of 4.
Formulation 3 composed of 83% salmon surimi (total weight basis), 3% salt,
2% WPC, 5% WS and 7% water resulted in a gel strength of 79.6 N.mm and
a fold score of 6. Since the objective was to maximize both the gel strength
and the fold score, formulation 3 was chosen to be the optimal formulation.
The characteristics of the gel obtained from this optimal formulation are very
close to those of commercial Kibun Hanpen-L (Boiled fish cake) (see Table
17). However, the gel strength of kamaboko produced from this optimal
formulation was 20% lower than that of Maruu Kamaboko Shiro, the second
target product. When compared to a control formulation (salmon surimi plus
3% salt), the optimal formulation caused a 35% increase in gel strength. It
also increased the fold score from 3 to 6. Formulations having a 6% WPC
consistently gave weak gels. Moreover, formulations consisting of 5% and
7% WS resulted in better fold score values than those containing 3% WS.
When the cumulative proportions of WPC and WS reached 11 % or higher
(i.e., formulations 7, 9 and 10), the corresponding fold score values were very
good but the gel strength values were low. The individual gel texture
improving effects of WPC and WS are demonstrated by formulations 11 and
12. These results confirm the findings of Kim and Lee (1987) who reported
the gel strengthening effect of starch and Lee et al. (1992b) regarding the
88
Figure 15. Combined effects of WPC and WS on salmon surimi gel fold score.
89
gel strengthening effect of W P C .
The amount of added water was calculated in a way that the final product
moisture content had to be around 70% (wet basis). The moisture contents of
the ingredients used are shown in Table 18. Although the addition of water
decreased the gel strength, it improved the product elasticity.
The optimal formulation (formulation 3) which was developed for surimi was
applied to surimi made from fresh herring. The product had a gel strength of
74.4 N.mm and a fold score of 6. These gel quality values are close to those
of Kibun Hanpen-L which is the lower quality target product. However,
because the composition of roe herring (i.e., fat content, protein content,
proportion of dark muscle) is different from that of pink salmon, the optimal
formulation for salmon is not necessarily the optimal formulation for herring.
Therefore, more research should be conducted to find the optimal formulation
for herring kamaboko.
90
Table 18. Moisture content of different products
Sample % Moisture
Salmon surimi 74.9
Herring surimi 73.6
WPC 5.1
WS 14.1
Salt 0.2
91
4.5 Conclusions
Kamaboko made from an optimized formulation consisting of 83% pink
salmon surimi (total weight basis), 3% salt, 2% WPC, 5% WS and 7% water
gave gel strength and fold score values close to those of Kibun Hanpen-L
(Boiled fish cake), which was used as a target product. When this formulation
was applied to surimi made from fresh herring, similar gel characteristics were
obtained.
Roe herring frozen at -40°C for one month and then at -45°C for forty days
maintained its gel making ability. Further storage for ninety more days
resulted in surimi gels of lower quality. This could be due to protein
denaturation due to frozen storage. However, when roe herring was frozen
at -40°C for one month and then at -83°C for 190 days, the gel making ability
was preserved.
Pink salmon seems to have a great potential to be used for surimi-based
products. Using pink salmon to make crab analogs, for instance, could be
more profitable than using it for canning. Because of its darker color, roe
herring could be successfully used in surimi-based products that do not
require a white color. The problem of its small size can be overcome by using
92
equipment that is designed for producing surimi from under-sized fish species
such as sardines. Using male roe herring to make value-added surimi-based
products is a better alternative than simply reducing it to fish meal.
93
C h a p t e r 5. S u r i m i v i s c o u s properties
Chapter 5
Viscous Properties of Salmon Surimi Paste
5.1 Introduction
The viscous properties of surimi are among the physical properties that need
to be thoroughly studied before new and improved processes for surimi-based
products can be developed. These flow properties of surimi pastes are of
particular importance during the pumping and the extruding operations that
occur during food processing.
Several researchers have studied the viscous properties of actomyosin
solutions. Nakayama et al. (1979) investigated the effects of incubation at
40°C and of the additions of Na P20 and MgCI on the viscosity of carp
4 7 2
actomyosin. The viscosity changes of croaker actomyosin solutions during
gelation were studied by Wu et al. (1985). They found that viscosity
decreased with temperature up to 31 °C. It then increased quickly, peaking at
36°C and then rapidly decreased. Owusu-Ansah et al. (1988) also observed
94
Chapter 5. Surimi viscous properties
similar trends of viscosity variation with temperature for sucker myosin. They
found that viscosity peaked at 43°C. Jimenez-Colmenero and Borderias
(1983) and Borderias et al. (1985) stated that the apparent viscosity of
proteins from frozen fish isolated in high ionic strength solutions can be used
as a quality index for frozen fish proteins. Groninger et al. (1983) used
viscosity measurement to assess changes in processed frozen fish muscle. A
test for the viscosity of surimi diluted (599% by weight) in a 3.5% salt solution
was used as a Japanese standard test for the quality evaluation of frozen
surimi protein (Hamann and MacDonald, 1992). The apparatus to be used for
this test is a Brookfield viscometer (Tokyo Keiki Type C). Scott et al. (1988)
found that the viscosity of Alaska pollock surimi, diluted in accordance with
the Japanese standard test, decreased with frozen storage time of the headed
and gutted Alaska pollock up to 128 days. The effects of incubation time and
temperature on the viscosity of a surimi paste (30% surimi, 2% salt, 1.2% beef
plasma protein and 66.8% water) were studied by Park et al. (1994). The
viscosity of the paste incubated at 5°C remained close to 40 Pas. When the
paste was incubated at 25°C, the viscosity increased to 80 Pa.s after 20 hours
of incubation. The viscosity of the paste incubated at 40°C for 30 minutes
increased to 65 Pa.s. However, the viscosity of the paste incubated at 60°C
for 30 minutes decreased to 10 Pa.s. The viscosity increases at 25 and 40°C
were attributed to intramolecular binding of myofibrillar proteins whereas the
95
viscosity decrease at 60°C was thought to be due to proteolytic degradation of
the myofibrillar proteins.
In the manufacturing of surimi-based products, surimi is mixed with
ingredients (i.e., salt, starch, etc.) in a silent cutter. The resulting viscous
paste is then pumped before being cooked and extruded (Wu, 1992). Hence,
it is important to study the viscous properties of surimi in the paste form.
The objective of this work was to study the viscous properties of a salmon
surimi paste having an optimum formulation for kamaboko making. The
results should help to clarify the flow behavior of this minced fish muscle.
The surimi used was the same salmon surimi described in Chapter 4. The
optimal formulation, which was selected in section 4.4.2, was used to make
the surimi paste for the Theological studies. As in section 4.3.2.2, partially
thawed surimi was cut in a vacuum cutter (Stephan, Model VCM-5, Columbus,
Ohio) at low speed for two minutes. Then 3% salt (total weight basis) was
added and the mixture was chopped at low speed for two minutes. Then 5%
96
wheat starch, premixed with 7% water, and 2% whey protein concentrate
(Alaco Surimi Plus) were added. This order was recommended by Wu
(1992). The ingredients were blended at low speed for two minutes. Finally,
mixing was performed under vacuum and at high speed for one minute.
There are several methods for the measurement of viscosity. The glass
capillary method is based on the measurement of the time for a known volume
of fluid to pass through a length of capillary tubing. The orifice viscometers
are simple instruments for quick measurements. The method consists of
measuring the time needed for a volume of fluid to flow through an orifice.
This method, however, is less accurate than the glass capillary method
(Bourne, 1982). Other instruments include the capillary viscometer, the falling
ball viscometer and the coaxial rotational viscometers. The latter type of
viscometer is very popular because it allows the study of the time-dependent
Theological behavior of the fluid, permits the continuous variation of the shear
rate, can handle Newtonian and non-Newtonian fluids and can be equipped
with a temperature control feature. For these reasons, the viscous properties
of the salmon surimi paste (kept in ice) were studied using a rotational
viscometer (Haake Rotovisko, Model VT500, Haake, Germany). A sensor
with a profiled cup and spindle, SVP, was used to reduce slippage. The inner
cylinder (rotor) has a radius of 10.65 mm and a height of 31.45 mm, while the
outer cylinder has a radius of 11.55 mm. Experiments were carried out at five
97
temperatures; 1,6, 11, 21 and 26°C. For each temperature, the paste was
put in the cup and kept there for ten minutes to come to temperature. Then,
the sample was sheared at shear rates increasing linearly from 0 to 400 s" in 1
ten minutes. After that, the shear rates decreased from 400 to 0 s" in ten 1
minutes. Finally, the sample was sheared again for ten minutes at rates
increasing from 0 to 400 s'\ Experiments were carried out in duplicate. A
personal computer to which the viscometer was interfaced was used to vary
shear rates and to record the corresponding shear stress values.
The Systat 1990 (Systat, Inc., Evanston, IL) software was used for the
regression analysis.
Figures 16 to 20 are surimi paste rheograms at temperatures of 1, 6, 11, 21
and 26°C, respectively. Each figure shows an increasing shear rate (ISR)
rheogram, a decreasing shear rate (DSR) rheogram and a second ISR
rheogram. For all the temperatures, the first ISR rheogram was not smooth or
reproducible. This could be due to the fact that the paste was initially at rest
or that the structure was being ruptured by the applied shear, possibly
98
600
• Increase D, 1st T=1 C •* •••
• Decrease D • • • ••••
550 4- A Increase D, 2nd
500 A .OS
A A A A A A A N A
450
A A A n
400 A *
A * °
350 4- A A
• A •
A O
A a
n°
£ . 300 A •
(0 A n D
A •
A C D
250 4-
200 4-
150 4-
100
50
H h
0 50 100 150 200 250 300 350 400
D [1/s]
Figure 16. Rheograms of salmon surimi paste at 1°C
99
650
+ Increase D, 1st T = 6C
a Decrease D
•••• ^ —6'
600
A Increase D, 2nd • ^rW 3
550
500 -0° .^AA"
450 • • A AAA
400
„ 350
2A A
10
Q.
A6 k
(0 300 AD
•
A
250 •
200
150 +
100
50
•+-
0 50 100 150 200 250 300 350 400
D [1/s]
100
650
• Increase D, 1st T = 11 C
• Decrease D ,fio n
600
A Increase D, 2nd
A
•o
A A
A A
A
550 AA
A A
A A A A
D A A A " A-A-AA-
A
D nO A A
D
A A
500 A A A
•
450 •• •
_D2 a A
8A
0°
O A
0
400
•a
• A
A A
350 AD
CO
(0
300
250 4-
200
150
100 4-
50
+
0 50 100 150 200 250 300 350 400
D [1/s]
101
650
• Increase D, 1st
• Decrease D
600 A Increase D, 2nd
• • •
550 A* U
•A A •
¥
*Aa = D
• A A A A A AA A
500 A A
D dO A A ^ A A
A
A
HO
A A^
450 +
A A A
H D
400
A "
A °
350
(0
0.
(0
300
250 +
200 +
150
100
50
+ +
50 100 150 200 250 300 350 400
D [1/s]
102
500
• Increase D, 1st T = 26C

• Decrease D
450
A Increase D, 2nd
400
••••
350 •
• •
A A
A
300 nrf 3
A A A
A A
n 0 0
AA A A A
A A A
A A A
co
D D D A A A A
A
<L 250 D A A A
A
W CP
• " A A
° A A
200
A° A
AA&V
•
150 4-
100
50
04 H h
0 50 100 150 200 250 300 350 400
D [1/s]
103
causing oscillations. After ten minutes of shearing, however, the DSR
rheogram and the second ISR rheogram became relatively smooth and
monotonic. These rheograms show that shear stress did not vary linearly with
shear rate. Therefore, surimi paste had a non-Newtonian behavior.
Moreover, a yield stress [shear stress at zero shear rate, or the shear stress
required for the material to flow (Tung, 1989)] was observed. This yield stress
increased with increasing temperature (for T < 21 °C). For all the
temperatures and for shear rates lower than 220 s" , the DSR rheograms and
1
the second ISR rheograms were very similar. However, at temperatures of
11°C and above and for shear rates higher than 220 s' , the shear stress
1
values of the second ISR rheograms became considerably lower than those of
the DSR rheograms. This hysteresis could be due to the loss of structure of
the paste that could occur at higher temperatures and especially after
shearing at higher rates for more than 25 minutes. It can be deduced that this
surimi paste behaved as a shear thinning fluid with a yield stress. The
existence of hysteresis despite the low ramp speed (40 s" / min) that was
1
used to increase and decrease the shear rate, demonstrates that the paste
rheology was time dependent and had a slow recovery.
Park et al. (1994) found that the viscosity of a Pacific whiting surimi paste
started to increase after 20 hours of incubation at 25°C, and did not increase
for an incubation temperature of 5°C. This is thought to be due to the
104
relatively low concentration of surimi (30% by weight) in the paste they
studied. (The surimi concentration of the paste used in this project is 83% by
weight.)
The DSR rheograms for the five temperatures used are shown in Figure 21
while the second ISR rheograms are depicted in Figure 22. Both of these
figures demonstrate that for the same shear rate, provided D < 200 s" , shear
1
stress values increased with temperature up to 21 °C. This is thought to be
caused by the protein-protein interactions which increased with temperature.
As suggested by Douglas-Schwartz and Lee (1988), actomyosin from salmon
could be less thermally stable than actomyosin from species living in warmer
waters. This increase of shear stress with temperature differs from the results
of Wu et al. (1985) who found that the viscosity of croaker actomyosin
solutions decreased with temperature up to 31 °C. This could be due to the
high concentration of surimi used in this study. Shear stress values at 26°C
were the lowest. This is possibly due to the fact that the paste started to gel
and the shear stress readings could have been influenced by slippage or by
the rupture of gel structure near the cup wall. It was attempted to run the
experiment at 31 °C but the spindle could not rotate because the required
torque was above the operating range for the rheometer. The gelling effect
on the sample was apparent when it was taken from the cup.
105
650
600 mA—A
A
A
W wXX*
MT~ •
.A A wX* •
Decreasing D
A A
**
550
500 X**
***A N
•P ••••
w * A A A
A
no 0
450 4- X* A A •
X* A
•
X A A • •
AA
400 X A
A
A D
A •
350
co
CL x
*A
A n °o • •••
CO
300 | A •
250 4- •
• T= 1 C
•
•
•••
200 - • T= 6C
• AT=
XT =
11 C
21 C
150 - -
• • T = 26 C
100 4-
50 4-
0 4 + +
50 100 150 200 250 300 350 400
D [1/s]
Figure 21. DSR rheograms
106
600
550 Increasing D
*XX * A A A
A A
A * A A £ : W a
* *°T
***X X A A A
x »H »rV
B
8 D
*\
500
X X A A A A
A BpD • # j;
w X . .A • • ^ • %
x x* *
• ° * •••••• * \
X A A
¥
450 4- x x> A
0*. •
AA
XX .A ••• ** D W
400 A • »
X AA D D ™ / •
X A A
350 A • • • *
A D D
ra
£L 300
••••
co • • ••••
•
250 +
•
200 • T = 1 C
• T = 6C
A T = 11 C
150 + X T = 21 C
• T = 26 C
100 4-
50 +
+ -+• +
50 100 150 200 250 300 350 400
D [1/s]
Figure 22. Second ISR rheograms
107
Since the first ISR rheograms were affected by the time dependency of the
paste, only the yield stress values were calculated for these curves. The
Theological data for the DSR rheograms and the second ISR rheograms were
fitted by the Herschel-Bulkley model and by the Casson model. (Although
only half of the results are shown, all the Theological data including the
replicate results were used in the curve fitting.) These models were explained
by Bourne (1982). The appropriate model should represent the flow behavior
of the fluid over the shear rate range to which the fluid will be subjected
during processing (i.e., pumping, mixing, etc.). It should
also accurately fit the experimental data. The following is a brief description
of the two models which were found to best represent the data.
Herschel-Bulkley (HB) model:
This model is also called the power-law plastic model. It is different from the
standard power-law model in that it includes a yield stress term. It is therefore
suitable for fluids having a yield stress. The equation of this model is:
S = S + KD
0
n
108
where S is the shear stress, K is a consistency coefficient, D is the shear rate,
n is the flow behavior index and S is the yield stress. The parameters of this
0
model can be determined by non-linear regression based on the least
squares method.
Casson model:
This model was originally developed to simulate the flow of printing inks
(Bourne, 1982). It is appropriate for some foods with a yield stress, such as
tomato catsup and molten chocolate (Bourne, 1982). The equation for this
model is:
S 05
= So 05
+ (Kc D ) 05
where Kc is a parameter. The parameters of this model can be determined by
first plotting the square root of shear stress against the square root of shear
rate and then finding the equation of the straight line which provides the best
fit of the data points.
Table 19 shows the curve fitting results of the second ISR rheograms. The
data were well fitted by both models (R > 0.94). The values of n, the flow
2
behavior index in the HB model, were close to 0.5. For both models, yield
109
Table 19. Curve fitting results of the second ISR rheograms
HB model T[C] R 2
So K n Note
1 0.963 219.235 11.822 0.534
6 0.988 253.365 8.302 0.597
11 0.948 309.752 12.65 0.505
21 0.957 319.549 17.117 0.477 D<220
26 0.970 134.562 8.137 0.513
Casson model T[C] R 2

So Kc
1 0.956 215.591 0.164
6 0.989 245.365 0.153
11 0.944 311.571 0.102
21 0.954 333.741 0.119 D<220
26 0.955 137.223 0.092
110
stress values increased with temperature up to 21 °C. The yield stress values
at 26°C were the lowest.
These trends were also observed for the curve fitting results of the DSR
rheograms (Table 20). These rheograms were also very well fitted by the HB
model and by the Casson model (R > 0.96). The curve fitting results of the
2
DSR rheograms were slightly better than those of the second ISR rheograms.
Figures 23 to 27 show the DSR rheograms along with the predictions of the
fitted models for the five temperatures used. The curve fitting results obtained
with the HB model demonstrate the existence of a yield stress that increased
with temperature up to 21 °C. The yield stress values of the second ISR
rheograms were higher than those of DSR rheograms.
The yield stress values, S 0 [Pa], predicted by the HB model for various
temperatures, T [°C], was fitted by the following equation:
So = C Ty
m
where C and m are the equation parameters, and T < 21 °C. A logarithmic
y
transformation of this equation yields the following linearized form:
111
Table 20. Curve fitting results of the DSR rheograms
HB model T[C] R 2
So K n
1 0.997 183.145 10.021 0.576
6 0.995 217.791 14.381 0.548
11 0.994 264.433 13.383 0.562
21 0.990 285.473 19.03 0.478
26 0.965 129.023 3.336 0.751
Casson model T[C] R* So Kc
1 0.996 184.311 0.196
6 0.994 227.624 0.228
11 0.994 267.594 0.215
21 0.988 310.236 0.137
26 0.972 103.338 0.274
112
150 4-
100 4-
50 4-
0 4 1 1 1 1 1 1 1 1
0 50 100 150 200 250 300 350 400
D [1/s]
Figure 23. DSR rheograms at 1°C
113
114
700
650
• Deacrease D T= 11 C
H.B. model
Casson model
600
550
500
450 +
400
n
SL 350
(0
300 | D
250
200 4/
150
100
50
— i —
50 100 150 200 250 300 350 400
D [1/s]
115
• Decrease D T = 21 C
H.B. model
Casson model
450 +
400 +
200 +
150 4-
•+-
50 100 150 200 250 300 350 400
D [1/s]
Figure 26. DSR rheograms at 21 °C
116
50 4-
0 -I 1 1 1 1 1 1 1 1
0 50 100 150 200 250 300 350 400
D [1/s]
117
log(So) = log(Cy) + m log(T)
The results obtained by fitting the above logarithmic equation to the S values 0
predicted by the HB model for both the second ISR and the DSR rheograms
are listed in Table 21. The fit was good for both the second ISR and the DSR
rheograms. The latter rheograms resulted in a better fit (R > 0.90). 2
However, for the pumping of surimi paste, the yield stress value that must be
overcome at the beginning of the process is the yield stress determined at the
first increasing shear rate rheogram. Table 22 lists these yield stress values
for different temperatures. These were the recorded shear stress values
corresponding to the lowest shear rate, 8.97 s~. 1

Generally, yield stress
increased with increasing temperature up to 21 °C. The lowest yield stress
value was observed at 26°C. Moreover, for each temperature the initial
yield stress was much higher than that of the DSR rheograms and the second
ISR rheograms.
The Herschel-Bulkley and Casson models depend only on the shear rate.
They do not account for the influence of temperature. The viscosity of
Newtonian fluids is found to vary with temperature according to the following
equation (Morgan et al., 1989):
118
Table 21. Results of the curve fitting of the yield stress
Rheograms Cy m R 2
Second ISR 215.406 0.130 0.874
DSR 179.205 0.148 0.915
119
Table 22. Yield stress values from the first ISR rheograms
Temperature, T Initial yield stress, S 0
(°C) (Pa)
1 371
6 462
11 457
21 522
26 329
120
ri = C exp (AE / RT)

T V
where r| is the viscosity, T is the absolute temperature, AE is the molar free

V
energy of activation in a stationary fluid, R is the universal gas constant and
C is a constant.
T
A new model, taking into consideration both the shear rate effect and the
temperature effect, is proposed. The equation of this model is assumed to be:
S =eD + gT f h
where S is the shear stress in Pa, T is the temperature in °C, D is the shear
rate in s" and e, f, g and h are the model parameters.

1
The form of this
equation derives from the fact that only the yield stress appears, in most
experiments, to be a strong function of temperature. Usually, the viscosity of
most fluids decreases with temperature (Streeter and Wylie, 1985). The
system used in this study, however, was likely affected by temperature
because of changes in protein-protein interactions.
For temperatures ranging from 1 to 21°C, the DSR rheograms and the second
ISR rheograms were fitted by the above 'shear and temperature (ST) model"
Table 23 summarizes the curve fitting results. The model parameters show
121
Table 23. Curve fitting results of Theological data by the ST model
Rheograms e f g h R
DSR 9.945 0.590 178.382 0.185 0.968
Second ISR 96.915 0.240 75.893 0.251 0.903
122
that shear stress increases with increasing temperature and shear rate.
Figure 28 shows the DSR rheograms as well as the predictions of the
fitted ST model. The second ISR rheograms along with the predictions of the
ST model are depicted in Figure 29. Curve fitting by the ST model was better
for the DSR rheograms (R > 0.96). The curve fitting was also better for shear
2
rates from 0 to 200 s" . The time dependency of the paste was mainly present
1
after shearing for 25 minutes. Therefore, the ST model should be applicable
for typical residence times of the paste as it is pumped between the mixing
tank and the extruder.
123
150 4-
100 4-
50 4-
0 -1 1 1 1 1 1 1 1 1
0 50 100 150 200 250 300 350 400
D [1/s]
Figure 28. DSR rheograms and the predictions of the ST model
124
600
50 100 150 200 250 300 350 400
D [1/s]
Figure 29. Second ISR rheograms and the predictions of the ST model
125
5.4 Conclusions
The viscous properties of a salmon surimi paste composed of surimi, salt,
whey protein concentrate and wheat starch were studied using a rotational
viscometer. The paste behaved as a time dependent non-Newtonian shear
thinning fluid with a yield stress. For the same shear rate, shear stress
increased with temperature up to 21 °C. This could be caused by the protein-
protein interactions that were facilitated by grinding the surimi with salt. The
yield stress values also increased with temperature up to 21 °C. The DSR
rheograms and the second ISR rheograms, after surimi had undergone shear
for sufficient time, were well fitted by the Herschel-Bulkley model and by the
Casson model.
A new model taking into account the shear rate effect and the temperature
effect has been developed. For temperatures up to 21 °C, the DSR rheograms
and the second ISR rheograms were well fitted by this new model. Predicted
shear stress increased with increasing temperature and shear rate.
Finally, it is important to mention that the paste used in this study was similar
to what is used in the commercial manufacture of surimi-based products (i.e.,
the paste was not first diluted with water as in most previous Theological
126
studies). The Theological information obtained should be useful for the
engineering design of the equipment needed for the pumping and extruding
operations that immediately follow the mixing process. However, the results
obtained here apply only for a salmon surimi paste having a specific
formulation and for temperature ranging between 1 an 26°C. A similar
approach can be followed to determine the rheological properties of other
surimi pastes.
127
Chapter 6. Conclusions and future work
Chapter 6
Summary of Conclusions and Proposed Future Work
6.1 Conclusions
Raman spectroscopy was successfully used to investigate the effects of
grinding with salt as well as setting and cooking on the protein structure of
raw surimi, surimi paste and surimi gels. The disulfide bond stretching
intensity increased slightly after setting and greatly after cooking at higher
temperature. These results explain the importance of the disulfide bonds and
the stages at which they occur. Secondary structure analysis showed that
setting caused a decrease in the a-helical content and an increase in the p-
sheet content. Cooking alone or preceded by setting resulted in a greater
increase of the p-sheet fraction and a further decrease of the a-helical
content. These findings suggest an unfolding of proteins allowing
hydrophobic interactions between exposed hydrophobic residues to take
place. Studies of gel properties also showed that cooking, whether preceded
by setting or not, resulted in a better surimi gel texture than the setting
128
treatment alone. In comparison with cooking alone, setting followed by
cooking resulted in a higher disulfide peak intensity and in stronger and more
elastic surimi gels.
Experiments on optimizing the setting and cooking conditions for kamaboko
making showed that setting at temperatures below 50°C followed by cooking
at 80°C and above resulted in surimi gels with the best gel texture
characteristics. The optimal processing conditions corresponding to
individually maximized gel strength, fold score and color, and minimized
expressible liquid were determined. Multiobjective optimization also gave the
optimal processing conditions resulting in a satisfactory combination of all
four response variables.
An optimized formulation composed of pink salmon surimi, salt, whey protein
concentrate and wheat starch has been developed. The final product,
kamaboko sausage, had gel strength and fold score values close to those of a
commercial surimi-based product. This formulation was successfully applied
to roe herring surimi. Freezing roe herring at around -45°C maintained the gel
making ability for up to seventy days. Freezing at -83°C preserved the gel
making ability for a longer period of time. However, this temperature is too
low to be used by the industry. Using male herring for surimi-based products
has a great potential in British Columbia.
129
A rotational viscometer was used to study the viscous properties of a paste
made from the formulation optimized in Chapter 4 and containing pink salmon
surimi, salt, whey protein concentrate and wheat starch. This paste had a
non-Newtonian shear thinning behavior with a yield stress. Apparent
viscosity and yield stress increased with temperature up to 21 °C. This could
be due to protein-protein interactions that increased the viscosity with
temperature. A new model taking into consideration the effects of shear rate
and temperature provided a reasonable representation of the rheological data
and demonstrated that up to 21 °C, shear stress increased with increasing
shear rate and temperature.
6.2 Proposed future work
For future studies, it is recommended that the mechanisms of surimi gelation
of other species be investigated using Raman spectroscopy. This method
could also be used to assess the changes of protein structure caused by the
frozen storage offish or surimi.
More work needs to be done in order to commercialize the use of herring, pink
salmon and Pacific whiting in the manufacture of surimi-based products. For
instance, new storage conditions for surimi should be determined in order to
130
prolong the frozen storage at reasonable temperatures (i.e., -20°C). Raman
spectroscopy could be used to investigate the changes in fish protein
structure due to frozen storage.
The viscous properties of raw surimi and of surimi paste should be studied
further using a measuring instrument, such as a capillary rheometer, that
could handle very viscous materials and hence could allow higher incubation
temperatures.
131
Nomenclature
Nomenclature
a redness index
b yellowness index
C whiteness index
CT model parameter of the Morgan model
D shear rate, s"1
e model parameter of the ST model
Ei expressible liquid, %
AE V molar free activation energy, J mol" 1
f model parameter of the ST model
F s fold score
g model parameter of the ST model
G s gel strength, N.mm
h model parameter of the ST model
•850/830 intensity ratio of the tyrosine doublet
K consistency coefficient of the HB model
Kc parameter of the Casson model
m parameter of the yield stress model
n flow behavior index
132
Nomenclature
N buried mole fraction of the buried tyrosine residues
N exposed mole fraction of the exposed tyrosine residues
Oi more resolved spectrum
P Likelihood function
Pc contracted vertex
PE expanded vertex
PH vertex with the highest objective function value
P* reflected vertex
R universal gas constant, J mol" K"

1 1
R 2
regression coefficient
s peak shape function
S shear stress, Pa
So yield stress, Pa
T temperature, °C
setting temperature, °C
x 2
setting time, min.
X3 cooking temperature, °C
X4 cooking time, min.
yi measured spectral data
Yj objective function of the vertex Pj
Y objective function of the SOM
Wj weighting coefficient
133
Nomenclature
WPC whey protein concentrate, %
WS wheat starch, %
Z multiobjective function
134
Nomenclature
Greek letters
a reflection coefficient
0 contraction coefficient
00 parameter of the SOM
0. parameter of the SOM
0H parameter of the SOM
0« parameter of the SOM
Y expansion coefficient
a standard deviation
viscosity, Pa.s
® convolution
135
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SURIMI-BASED PRODUCT DEVELOPMENT AND VISCOUS

PROPERTIES OF SURIMI PASTE.

Engineering Degree, School of Agricultural Engineering, ESIER, Tunisia, 1988

A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF

THE REQUIREMENTS FOR THE DEGREE OF

THE FACULTY OF GRADUATE STUDIES

(Department of Chemical Engineering)

We accept this thesis as conforming to the required standard

THE UNIVERSITY OF BRITISH COLUMBIA

© Moez M. Bouraoui, 1995

Department of CM&MUU £NfyMg£#WQs

cooking (86°C) or setting followed by cooking. The intensity of the peaks

assigned to disulfide bond stretching vibrations increased considerably in the

were subject to setting alone. Secondary structure estimation based on the

amide I band indicated a change from predominantly a-helical structure in raw

surimi to similar proportions of a-helical and anti-parallel p-sheet after setting.

A further increase in anti-parallel p-sheet and decrease in a-helix content

occurred during the kamaboko stage. The intensity of C-H stretching

vibrations of the aliphatic residues decreased after salting, setting and

Response surface methodology was used to determine the optimal setting

and cooking time and temperature conditions resulting in a maximized gel

strongest influence on the gel quality characteristics. Level set programming,

better than those generated by the Simplex technique.

Using a formulation composed of pink salmon surimi, salt, whey protein

concentrate and wheat starch, an optimal final product, kamaboko sausage,

was developed. In addition, when this formulation was applied to herring

commercial surimi-based product. The effects of frozen storage conditions of

further increased the GMA period.

The viscous properties of a salmon surimi paste were studied using a

rotational viscometer. The paste behaved as a shear thinning fluid with a

yield stress that increased with temperature up to 21 °C. Viscosity also

increased with temperature up to 21 °C, possibly because of protein-protein

interactions. The Theological data were reasonably well represented by a

1.1 Surimi manufacturing 3

1.2 Production of surimi-based products 6

1.3 Freeze denaturation of fish proteins 7

1.3.1 Moisture phase changes 9

1.3.2 Fish lipids 10

1.3.3 Enzymatic denaturation by TMAOase 10

1.4 Thesis outline 11

Whiting Surimi and Gels Using Raman Spectroscopy 13

2.2 Materials and methods 18

2.2.2 Raman spectroscopic analysis 19

2.2.3 Measurement of surimi gel properties 22

2.3 Results and discussion 25

2.3.1 Disulfide (S-S) bond stretching vibrations in the

450-600 cm' region

2.3.2 Backbone and side chain vibrations in the

610-2000 cm" region1

2.3.2.1 The tyrosine doublet near 830 and 850 cm" 1

2.3.2.2 Secondary structure estimation from the

amide I and III bands 35

2.3.3 C-H Stretching vibrations in the 2500-3400 cm" region

Chapter 3: Optimization of the Setting and Cooking Conditions for

Kamaboko Making from Pacific Whiting Surimi 46

3.1.1 The Nelder-Mead simplex optimization method 47

3.1.3 The Level Set Programming (LSP) method 50

3.2 Materials and methods 51

3.2.1 Gel strength (compression test) 53

3.2.2 Fold score 53

3.2.3 Gel color test 54

3.2.4 Expressible liquid measurement 54