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Tofu: technological and nutritional potential

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 Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017

TECHNICAL ARTICLES
Tofu: technological and nutritional potential
Anirban Dey1, Rasane Prasad1,2*, Sawinder Kaur1, Jyoti Singh1, M.D.Luwang1
1
Department of Food Technology and Nutrition, Lovely Professional University,
Jalandhar 144411 Punjab, India;
2
Centre of Food Science and Technology, Banaras Hindu University,
Varanasi - 221 005, Uttar Pradesh, India
*E-mail: rasaneprasad@gmail.com

Abstract soymilk or soy juice. Soy milk should be sterilized

Tofu is popularly consumed for its health benefits
and nutrition. It has been increasingly used in
numerous culinary dishes, replacing dairy products
due to comparatively low cost and high protein
bioavailability. The physico-chemical, functional,
microbiological and sensory quality of tofu varies
due to soybean cultivars, coagulants and varied
techniques used. The mechanized manufacturing of
tofu is still at its infancy and most of the production
is confined to cottage level. Lack of well-defined
standards is hampering its utility. Current paper
reviews various aspects of tofu’s history, gelation
mechanism, manufacturing processes, types,
quality, mechanization, physical, chemical and
sensory properties, health benefits and shelf life, to
establish its importance in the global food market
and to device a possible roadmap for its future in
human diet.
Keywords Tofu, Quality, Mechanisation, Shelf life,
Tofu gel
Introduction
Tofu also known as soy curd is generally
obtained by coagulating soymilk. Coagulants such
as citric acid (Murugkar 2015) and CaCl2 or CaSO4
(Oboh 2006) Calcium sulphate (CS), Nigari (MgCl2)
and glucono-d-lactone (GDL) are majorly used in
Japan for the preparation of tofu (Kohyama et al
1995) During production of the tofu, coagulant
forms a soy protein gel matrix which helps in
entrapping water, lipids and constituents present in
TARs
by boiling for 2 min or an equivalent method which
has a better sterilization effect. The packaged tofu
can be sterilized by heating at 90°C for 45 minutes
or an equivalent method to sterilization. Stored tofu
must be refrigerated or dipped in chilled potable
water in water bath which needs to be changed
periodically. Pasteurization and packaging helps to
extend the storage life of tofu by a year. The
coagulation of soy protein is a complex interaction
as there is a complex interrelationship between the
contents viz. variety of soybean, cooking temperature
of soymilk, pH, coagulants used, solid content and
coagulant amount and time (Cai and Chang 1998).
The texture and flavor of the tofu differ with the use
of different coagulant. However, generalized texture
of tofu should be smooth, firm and coherent
whereas, hard and rubbery texture is not accepted
(Poysa and Woodrow 2004). Tofu standards are
nearly non-existent and a concentrated review of tofu
is much needed to investigate various types of tofu.
This paper, thus reviews various aspects of tofu such
as manufacturing processes, gelatinization
mechanism, mechanization, types of tofu, chemical
composition, health benefits, microbiology,
functional properties and sensory aspects for better
understanding of tofu.
Historical aspects of tofu
It is believed that the soybean has originated
from China nearly 4000-5000 years ago and was
used as a popular product in Chinese cuisine
(Watanabe 1997). China was the world’s largest

the matrix to form curd, further pressurized to form
solids (Cai et al 1997; Oboh 2006). According to
Japan External Trade Organization (JETRO, 2011),
soybean curd (tofu) should be manufactured from
8
producer of soybeans and its exports till 1954 until
United States succeeded them (Penarananda 1999).
China being the birth land of tofu it gave the name
originally called as doufu, documented by Tao Ku
 Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017

in 950 A.D. The term tofu was however, coined by by numerous scientists in succession. Following are
Japanese in early 1182. During the developmental some popular theories of soy gel formation,
period of tofu in 14 century people of Japan gave however; the presently understood theories still
various nicknames such as shirakabe, shirokabe and suffer from numerous drawbacks.
okabe. Though soybean is not technically a grain,
Kohyama et al (1995) proposed a popular
according to Chinese culture soybean was one of
soy gel formation theory. The gelation mechanism
the essential grains amongst the five sacred grains
of tofu as hypothesized by them follows two steps
including rice, wheat, barley and millet. The creator
denaturation of protein and hydrophobic
of tofu Liu An prepared soybeans similar to grains
coagulation. The first step is to expose the protein
followed by drying, mashing and boiling with
molecules residing in native state, consisting of
addition of sea salt. This lead to seasoning of the
hydrophobic regions, to the heat for denaturation.
slurry and also acted as solidifying agent to form
The denaturation of soybean protein produces
curd. Since then, soybeans were processed into tofu
negatively charged ions. In the second step, geneses
in 200 B.C. using sea salts. According to Fan (2014)
of protons are triggered by glucono-delta-lactone
tofu has originated from a county known as
(GDL) and calcium ions helps in neutralizing the
Shouxian in the Province of Anhui located in
net charge present in the protein. This results in
Southeast China. Shouxian civilization was formerly
better hydrophobic interaction between the
known as Shouchun or Shouyang at the time of
molecules of neutral proteins and induces
Zhou Dynasty 203 B.C. due to various historical
aggregation of protein molecules finally leading to
and cultural occurences Shouxian was awarded the
formation of soy curd. Figure 1. represents the
title of historical and cultural city of China. In
Kohyama’s mechanism of gel formation in tofu.
Shouxian tofu was prepared by coagulating smashed
Summarily, the coagulant addition neutralizes the
soybean along with water and formed into curdling
net charge on the denatured proteins and the
box. Although the world’s largest population
hydrophobic interactions induced aggregation of the
consumed it, tofu appeared in the international
denatured soy proteins this results into formation of
market rather later, it first appeared in the
tofu gel matrix (deMan et al 1986). However, this
supermarkets in America only during 1980’s.
process of denaturation is still in progress and
Gelation mechanism of tofu continues during the storage, leading to syneresis
in tofu. Murekatete et al (2014) studied effect of
Various gel formation theories are proposed

TARs

9
 Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017

coagulants on gelation behavior of tofu. They concentration of protein. However, in yet another
reported that the salt induced gels had high storage study on 11S and 7S proteins by Kohyama and
modulus (G’) and shorter gelation time. Nishinari (1993) the gelation time was found to be
decreased with increase in 7S globulins in presence
Soybean proteins are made up of two major
of GDL. Thus, it could be stated, though 7S proteins
fractions 7S (â-conglycinin) and 11S (glycinin). 7S
are slower to form gel however, the rate of gelation
subunit is a soluble protein while, 11S exists in a
by 7S globulins could be increased in the presence
particulate form (Peng et al 2016). It is determined
of appropriate coagulants. In addition to this 7S
that the rate of gelation of 7S globulins is much
proteins could be coagulated at lower concentration
slower than that of 11S proteins (Kohyama and
as compared to 11S fractions. Thus, it is evident that
Nishinari 1993). Amongst soy proteins, 11S
7S proteins are mainly responsible for rate of
globulin fractions are the major protein determining
gelation, while the hardness is influenced by 11S
the hardness of soy gel (tofu) (Saio and Watanabe
protein fraction of soy protein. Moreover, Nakamura
1978). Kohyama et al (1992) studied the rheology
et al (1985) studied 11S fraction of soy protein in
of gel formation by soybean 11S protein in
minor fractions namely A 1, A 2, A 3, A 4 and A 5
presence of a coagulant. They observed the gelation
subunits. A 3 subunit was found to increase the
curve in relation to temperature and rate order
hardness of the gel, concluding, that the hardness
kinetics. They found that the rate of gelation
of the soy gel is proportional to the percentage of
increases with gelling temperature and is influenced
A 3 subunit present in the soy cultivar. Further,
TARs

by the coagulant such as glucano-ä-lactone. They

Nishinari et al (1991) studied the effect of A5 subunit
also reported that while latent time of the other
of soy globulins on the gel characteristics of tofu.
protein based gels are dependent on the protein
They concluded that the soy gel prepared with A5
concentration, the soy gel is independent of the

10

negative cultivars of soybean yielded harder tofu gels
as compared to their positive counterparts.
However, the model of soy gelation
described by Kohyama et al (1992) suffers from a
major drawback, as it does not take into
consideration lipids present in the soybean in the
gelation process. As compared to proteins, lipids
are not well dispersed in the gel interspaces. As the
coagulant is added, the lipid gets incorporated in
the gel matrix (Yamano et al 1981).
Clark et al (2001) illustrated another view of
gel formation deviating from the particulate gel
theory. They hypothesized that the intermolecular
electrostatic repulsion plays a major role in favoring
the formation of gel, which is less like the particulate
gel. The repulsion is believed to be due to surface
charges and is influenced by the nature of the
particle, pH and concentration of the coagulant.
This hypothesis stated the need to study the
interative effect of particulate matters like phytic
acid, polyacid ions, proteins and lipids
Concomitantly, Shun-Tang et al (1999)
proposed another model for soy gel formation
which integrates soy protein, lipids and small
charged molecules mainly phytates and polyacid
ions. In addition to the Kohyama’s model, it
proposes, after the 11S protein has been coagulated
in presence of Ca 2+ ions, as the coagulant is
added, smaller molecules prevents the protein-lipid
interface from coagulating. The lipid globules are
now wrapped with particulate 11S rich proteins
micelles and later with soluble 7S rich particles.
During this process the lipids form specific oil
bodies and coalesce and get coated with oleosins
(structural proteins). This micelles appears
sandwiched, many such sandwiched protein-lipid
interface forms soy-milk gel matrix. Figure 2. shows
the formation of soy gel according to Shun-Tang’s
theory. This model is further replicated to
understand yogurt and cheese matrix (Grygorczyk
et al 2014; Perg and Guo 2015; Ringgemberg et al
2013). Further improvement in the model is
however thought to be necessary, as the soy gel
matrix is thought to bear a particulate gel structure
and the gelation occurs when protein-lipid complex
coagulates due to intermolecular crosslinking. This
complex appears as particle type proteins clustered
together (Nik et al 2011).
11
Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017
Types of tofu
In last few decades efforts have been made
to introduce numerous varieties of tofu such as
silken tofu, firm tofu, dried tofu, fermented tofu and
low fat tofu. A schematic flowchart for the
manufacturing of different types of tofu is depicted
in Figure 3. A brief description is provided below.
Firm tofu:
Firm tofu also known as “Cotton tofu” is
prepared from soymilk, coagulated with synthetic
or natural coagulant and pressed through cheese
cloth. It is partially coagulated and pressed to
remove excess water and the moisture in the tofu
ranges from 76-81% (deMan et al 1986). It is firm
because excess of moisture is squeezed using heavy
weights. Natural coagulant extracted from fruits viz.
Phyllanthus acidus (gooseberry), Tamarindus indica
(tamarind), Citrus limonum (lemon), Garcinia indica
(garcinia) and Passiflora edulis (Passion fruit) and
synthetic coagulant viz. CaSO4 (Calcium sulphate)
and MgCl 2 (Magnesium chloride) are used for
coagulation. The soymilk is prepared from cleaned
and soaked soybean for 12 h at room temperature.
Soybean slurry is obtained from ground soybean
with water (1:8) and indirectly heated at 85°C for
45 min with constant stirring to avoid charring. The
hot slurry is clarified with the help of cheese cloth
and soymilk is obtained. The soymilk is heated up
to 95°C for 5 min, 0.2% synthetic and/or 2% of
natural coagulant is used, stirred for 5 min and
allowed to coagulate for 15 min. Coagulated mass
is further pressed in tofu mould lined with double
cheese cloth for 20 min at 1 kg pressure initially and
0.5 kg for next 20 min. Finally, tofu is stored at 4°C
(Rekha and Vijayalakshmi 2010).
The application of bittern (MgCl2·6H2O) was
studied by Li et al (2013) for tofu preparation using
W/O emulsion of bittern solution and soybean oil
at ratio of 1:1.5 or 1:1 (w/w). Cleaned soybeans
were soaked in distilled water (3 times more) at
25°C for 12 h. Water was drained and beans were
ground with distilled water (1:6 w/w) using
automatic separation grinding machine. Soymilk
with a total solids ranging from 7.8 to 8.2% was
obtained and then heated in ohmic heating kettle
at 100°C for 15 min. It was then rapidly cooled to
TARs
85°C, around 500 ml of soymilk was coagulated
by pouring into coagulant (bittern solution or W/O
emulsion) chamber. For proper dispersion and
 Indian Food Industry Mag
TARs Vol 36 No 3, May-Jun 2017

12

release of coagulant in case of W/O emulsion, high-
speed shearing machine was used at a speed of
13,000 rpm. Total solidification was observed at
85°C for 30 min. It was further poured in perforated
moulds (10cm x 7cm x 8cm) and pressed for 30 min
at a constant pressure of 2,200 g to produce tofu.
Silken tofu/ soft tofu:
The bean curd generally obtained for the soy
milk is coarse and irregular in texture whereas,
silken (Kinugoshi) tofu as the name suggests is fine
and smooth in texture (Beddows and Wong 1987).
Silken tofu is a variety which is produced by
solidifying all the soymilk providing a soft and fine
texture to silken tofu. The moisture content of soft/
silken tofu is about 87-90% (Wang et al 1983).
Yang and James (2013) manufactured silken
tofu by the soaking method, 55 g of soybeans were
soaked in 200 g of deionized water at 15°C for 16-
17 h. Drained soybeans were rinsed and 400 g of
deionized water was added for blending and
obtaining of soy slurry. The slurry was cooked at
98°C for 15 min. In the dry method used by Evan
et al (1997) raw soybean of 60 g was ground and
blended with 400 g of boiled water. Extraction of
soymilk from both the method was by manual
centrifugal juice extractor. Concentration of
coagulant used ranged from 1.5 to 5.0 g per kg.
Then 220 ml of soymilk was poured in a plastic
container and cooled to room temperature, 7 ml of
coagulant was added and stirred vigorously and
thoroughly. The container was sealed and indirectly
heated at 85°C for 35 min to complete the curd
formation, lids used for sealing was provided with
holes to escape the gases and silken tofu was stored
at 4°C.
Dried tofu:
Among all the types of tofu, dried tofu is the
most firm tofu having moisture content below 76%.
The moisture content of dry tofu is low as the
soybean curd is broken vigorously to exclude extra
water before pressing (Cai and Chang 1997).
The dried tofu is prepared by steeping raw
soybeans in 6-folds of water at 21-22°C for 8 h.
After soaking, excess water is drained and hydrated
soybeans are ground in high-speed grinder which
is clarified by centrifugation to obtain soymilk. For
the preparation of dried tofu, soymilk was boiled
for 20 min with 1 g of antifoaming agent and rested
for 3 min at 96°C. Nigari (3%) (MgCl2) is added and
13
Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017
allowed to stand for 10 min for the formation of
curd. The curd is stirred for 30 s and again
coagulated for the second time. Further, soy curd
is transferred to wooden mould lined with cloth (25
cm x 25 cm x 7 cm) and pressurized at 34.8 g per
cm2 up to 30 min (Cai et al 1997).
Sufu (Fermented tofu):
Sufu is also known as fermented tofu. The
tofu is fermented with various strains of
microorganisms viz. Actinomucor elegans, Mucor
racemosus, Mucor sufu, Mucor dispersus, Mucor
racemosus, Mucor wutuongkiao, Aspergillus sp.
The final product quality depends upon the type of
strains used for the fermentation. Chou and Hwan
(1994) used Aspergillus taiwanensis (CCRC 31159)
and Aspergillu selegans (CCRC 31342) for the
development of sufu. Tofu was sliced into pieces
and weighed (25 g) having 70% moisture content
which was soaked at 95°C for 6 min to prevent
from putrefactive organisms and dried in laminar
air-flow hood for 2 h. Inoculation of test
microorganisms was done by spraying 0-2 ml of the
culture on the surface of each piece. Pieces were
incubated for 48 h at 30°C and maintained at 97%
relative humidity. Finally, ageing of sufu was carried
out for 75 days at 25°C, mixed with brine solution.
Red sufu was prepared by Han et al (2001) soymilk
was coagulated using salt to prepare tofu. Then
diced tofu was sprayed with pure culture of
Actinomucor elegans As3.227 and incubated at
28°C for 48 hours at relative humidity of 90%. Salt
concentration was increased up to 16% by
continuous salting for 5 days and stored for ripening
for 2 months in an enclosed bottle dressed with a
mixture of kojic red rice (Japanese red yeast rice),
sugar, chiang (wheat-based miso), an alcoholic
beverage and spices.
Low fat tofu:
The consumers of the modern era are craving
for health foods with low fat. Tofu being an
excellent source of protein holds a good chance for
being a health food substitute by minimizing fat
consumption. Byproduct such as soy flour is
obtained from the soy oil cake, which is low in fat
content and have high amount of proteins with good
TARs
biological value (Kellor 1974). Shin et al (2014)
reported that supercritical CO2 extracted soy flour
has potential for production of low fat tofu that is
similar to full fat tofu. Fat content of the flour is
 Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017
reduced from 19.5 to 7.1%, leads to decrease in
soymilk viscosity from 50 to 40 cp, however, the
overall yield is better in low fat tofu (69.7%)
compared to full fat tofu (60.8%). The reduction in
fat level does not influence the textural and sensory
properties of tofu significantly. However, low fat
tofu still scored relatively low due to its roasted
appearance and nutty flavor as compared to full f
at tofu.
Specialized varieties of tofu:
Pea tofu is prepared by blending soybean
and pea using glucono-d-lactone (GDL) as
coagulant. Peas are soaked at 20°C for 17 h and
are mixed with soybean in the ratio of 1:1. The
soybean and pea milk is heated at 95°C for 15 min
using GDL and CaSO 4 at 0.3% and 0.1%
respectively at 95°C for 45 min for solidification
(Fengying et al 2009). For the production of dreg-
free tofu, soybeans are ground finer (twice) and
homogenized at pressure of 400 Kg per cm 2
maintaining the particle size of soymilk below 8
µm. Huai et al (2003) prepared dreg-free tofu where,
the concentration of soymilk is kept to 12%, 3.5%
coagulant was used and bittern : gypsum was used
in the ratio of 4:6. Soymilk was maintained at 80°C
during coagulation. Cheng and Li (2010) analyzed
the gelation properties of control and okara
(Abelmoschus esculentus) containing tofu
coagulated with CaCl2. The okara containing tofu
was darker in colour, had an increased yield and
water holding capacity with soft texture than the
control tofu.
Sufupehtze is manufactured from white
whole soybeans. Parameters such as particle size
of soy slurry, total solid content, coagulant type,
and coagulant concentration affect the quality of
Sufupehtze. Optimum sufupehtze is prepared with
maintaining 12% TS, using a blend coagulants of
gypsum and brine (5:5) added at the rate of 3%
and heating at 80°C provides sufupehtze that is
similar to the traditional product (Ran and Kan
2012).
Factors influencing the quality of tofu
Variety of soybean seeds:
Several types of soybean genotypes from
TARs
different locations were evaluated by Bhardwaj et
al (2004) to determine the effect on oil and fatty
acid content of the tofu. The different genotypes
14
used were BARC-8, BARC-9, MD86-57788, Enrei,
Hutcheson, Nakasennari, S90-1056, Suzuyutaka,
Ware, York, V71-370 and V81-1603 among which
Hutcheson resulted in higher oil content (24.0 g/100
g) in tofu and its total saturated fatty acid content
was 3.80 g/100 g in tofu whereas, BARC-8 and
BARC-9 showed lower oil content of 15.8 and 11.3
g/100 g respectively. Seeds grown in different
locations resulted in variable content of total
saturated fatty acid in tofu.
Effect of coagulant:
The properties such as yield, texture, color
and sensory attributes are influenced by different
coagulants used to manufacture tofu. Obatolu (2008)
studied the effect of calcium sulfate, epsom salt,
lemon juice, alum and top water of fermented maize
for coagulating soymilk. Yield of tofu was highest
with calcium sulphate (565.7 g) and lowest with
alum (442 g). Textural properties of tofu such as
hardness ranged from 525.6 to 1008 g, chewiness
was between 2.3 to 4.5 kg and brittleness varied
from 1035 to 1482 g. The yield of tofu was around
404 g using GDL (Jackson et al 2002). Tseng and
Xiong (2009) investigated rheological properties of
inulin incorporated, GDL coagulated silken tofu.
The inulin was incorporated from 1 to 4% which
showed increase in hardness from 115.8 to 137.4
g, cohesiveness decreased from 0.86 to 0.82,
percentage deformability ranged from 5.19 to 8.25
and there was an increase in rupture force from
424.3 to 487.9 g. Coagulants such as acids (lactic
acid, acetic acid and rice vinegar) and salts (CaSO4
and MgSO4) used at industrial scale (18 L coagulant
used in 45 L soymilk) had yield ranging from 178.85
to 186.94% and 204.58 to 221.68% in acid and
salt, respectively (Sidar et al 2011).
In the recent years, application of chitosan
in different fields had led to development of various
applications including edible packaging from
chitosan which is highly successful in the current
era. Study was done using chitosan as a coagulant
to improve the shelf-life of tofu in which six different
types of chitosan were selected on the basis of
molecular weight (1106, 746, 471, 224, 28 and 7
kDa) using various treatments. Optimum treatment
used for coagulation of soymilk 1% chitosan solution
in 1% acetic acid at 80°C for 15 min. A
comparative storage study was done between CaCl2

coagulated tofu and chitosan-tofu at 7°C for 7 days,
revealed chitosan-tofu exceeds the shelf-life by 3
days than that of CaCl2 coagulated tofu (No and
Meyers 2004).
Effect of enzymes :
Yasir et al (2007) analyzed the effect of
transglutaminase (tga) enzyme on texture of tofu.
The fracture force of control tofu was over 30N and
the tga-tofu had fracture force less than 25N.
Transglutaminase treated tofu with 1000 ppm
showed facture force of 25 N and for tofu treated
with 5000 ppm transglutaminase it was above 30N.
This showed that enzyme content has a significant
effect on the texture of tofu.
In another study, commercial
transglutaminase was used to induce the gel
formation in tofu, the addition of transglutaminase
had significant effect on the texture profile of tofu
as a function of pH and NaCl. Hardness and
springiness of tofu was highest at pH e”6.4,
whereas, cohesiveness had no significant changes
in all the pH range tested. Addition of sodium
chloride (NaCl) only affects the hardness of
transglutaminase induced tofu and no effect on
springiness, gumminess and cohesiveness at an
optimum temperature of 37°C was observed (Tang
et al 2007).
Effect of process parameters
The process parameters such as thermal
treatment with sodium bicarbonate (NaHCO3), total
solid content of soymilk, time of stirring, moulding
have a significant effect on the texture and quality
of tofu. The softness of tofu (3.2 N) increased with
the increases of blanching time whereas control tofu
had a hard texture (7.8 N). Ten minutes ( 10 min)
of thermal treatment with 1% sodium bicarbonate
treated soybeans produced smooth tofu obtained
from soymilk having 7° Brix solid content. Stirring
time and moulding both affects the yield of tofu, 5
s stirring of soymilk after addition of coagulant
resulted in higher yield of 22.5 g/100 ml than 20 s
stirring (19.16 g/100 ml) whereas, in moulding,
1000 g weight for 15 min followed by 500 g results
in 22.6 g/100 ml yield having regular texture of 4.2
N hardness than only 1000g weight for 30 min
resulting in 22.3 and 7.8 N in yield and hardness,
respectively (Rekha and Vijayalakshmi 2013).
15
Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017
Mechanization of tofu making
In the soybean products and tofu production
degree of industrial automation is limited especially
in the section of dried tofu production. The
manufacturers generally runs, handicraft workshops,
mechanized and industrialized processes are few.
Huang and Dong (2010) designed pressing
equipment for the dried tofu production for
industries. Virtual prototyping technology was used
for the designing, three-dimensional modeling by
Pro/E software and AD-AMS software helped to
check feasibility analysis of kinematics and
dynamics. It helped in optimizing the design,
analysis and cost control which had great effect on
mechanization and industrialization.
Kuan (1998) invented a form system for fast
and hygienic production of tofu. The form system
comprised of a frame, top and bottom plate, form
grid and border having perforations. Tofu was
produced by fixing the frame on the bottom plate
and horizontally positioning the lower form grid
followed by spreading the lower gauge cloth. Then
raw material is poured uniformly on the lower
gauze cloth and an upper gauze cloth were used
to cover the raw material from the top. The system
was then packed using an upper form grid and top
plate on it. Finally, pressure was applied on the
system and the final product was recovered by
dismantling all the parts of the form system.
For the production of tofu in pilot scale,
automated tofu processing systems are used
consisting of soaking tanks, conveyors, feeder,
soybean grinder, milk separator, storage tank of raw
milk, antifoam feeder, cooker equipped with direct
steam injection, coagulant mixing vat consisting of
stirrer, filler, tank filled with coagulant, tofu trays
and presses with an air cylinder. To run the process
automatically, the system is equipped with a
computer to keep a check on the various process
parameters. The machine has a mass production
capacity of approximately 240 kg tofu per hour (Cai
et al 1997).
Chemical composition
The chemical composition of tofu varies to
great extent depending upon soybean type,
TARs
sprouting of soybeans and processing as shown in
Table 1.
 Indian Food Industry Mag
TARs Vol 36 No 3, May-Jun 2017

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TARs

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 Indian Food Industry Mag
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Study conducted by Murugkar (2014)
observed, that germination of soybean influences
the composition of tofu. Soymilk was obtained from
both sprouted and non-sprouted soybeans,
coagulated with 3% CaSO4 at 80°C. Tofu from
sprouted soybean varieties showed 13% increase
in protein and decrease in fat (12%), trypsin
inhibitor (81%) and phytic acid (56%).
Health benefits of tofu
Numerous researchers suggest several health
benefits of tofu. Consumption of soy food has been
studied to lower certain chronic diseases like
cancer (hormone-dependent), cardiovascular
diseases and osteoporosis. Soybean protein showed
hypocholesterolemic and soy isoflavones possesses
anti-atherogenic effect (Carrol 1991). It was also
observed that the intake of soy protein effectively
decreased serum concentration of total cholesterol,
LDL cholesterol and triglycerides as compared to
animal proteins (Potter 1998). Isoflavones, aglycones
and proteins present in tofu have antioxidant
properties protecting from lipid oxidation. Though
processing of soybean into tofu reduces the amount
of isoflavones from 1267 in soybean to 455 mg per
g in tofu nonetheless, this amount present is
sufficient enough to be provide potential health
benefits (Jackson et al 2002).
Functional tofu
The live microorganisms that when
administered in adequate amounts could confer a
health benefits on the host, these are known as
probiotic bacteria (FAO 2001). Probiotic bacteria
exhibit various health benefits such as stabilization
of gut microflora, decomposing indigestible
saccharide and produce organic acid. Zielinska et
al (2015) developed probiotic tofu using
Lactobacillus casei at the rate of 9.26 log CFU/g,
having an incubation temperature of 37°C for 2 h
using CaSO4 as a coagulant. The Bifidobacterium
animalis, Lactobacillus and other lactic acid bacteria
was maintained at, 103, 108-109 and 109-1010 CFU/
g, respectively, for 15 days at 4°C and acceptability
was checked by sensory which was quite
acceptable with high count of probiotic bacteria.
Similarly, growth and bioactivity of
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probiotics in tofu using various microorganisms
such as Lactobacillus acidophilus FTCC 0291,
Lactobacillus bulgaricus FTCC 0411, Lactobacillus
fermentum FTD 13, Lactobacillus casei FTCC 0442
18
and Bifidobacterium bifidum BB12 was studied by
Keat-Hui et al (2008) All these microorganisms were
screened for their á-galactosidase activity over 24 h
out of these L. bulgaricus and L. fermentum
performed best. For the production of probiotic tofu,
L. bulgaricus and L. fermentum was inoculated on
tofu and stored for 9 days at 4 and 25°C
respectively, during the storage period concentration
of both the probiotic bacterium were maintained at
10 6 CFU/g. During storage tofu showed
preservative effect, increase in organic acid
concentration, decrease in pH and angiotensin-
converting enzyme inhibition.
Packaging of tofu
Tofu is a perishable product and is prone to
physico-chemical and microbial spoilage. Therefore,
its packaging is necessary to provide protection from
spoilage to maintain the quality, freshness, consumer
appeal and sales of tofu. Cha et al (2003) analyzed
the effect of nisin coated polyethylene films for the
inhibition of Listeria monocytogenes. Shelf life study
of tofu for 30 days at 5°C incorporated with L.
monocytogenes Brie-1 packaged in normal
polyethylene pouches and nisin-coated polyethylene
films was carried out. The study revealed that the
polyethylene film having 1000 IU/g concentration
had a reduction of counts from 2.74 x104 to >101
after 30 days than other concentrations of 0, 100,
500 IU/g. The study shows that packaging of tofu
in nisin film can overcome the problem of survival
of L. monocytogenes.
A comparative study between air and
modified atmosphere packaged (MAP) tofu was
carried out for a period of 28 days. Gas mixture
containing CO2 and N2 was used. Headspace gas
was analyzed in the packaged samples. Low levels
of oxygen was present in vacuum compensation
package than in flushed samples after 10 days of
storage, whereas, the air packed tofu samples had
no change in the O2 levels. In short term analysis,
the CO2 level decreased to 15 and 20% for vacuum
compensation and flushed samples, respectively and
increased by 4% in air packed samples. The O2
simultaneously increased in vacuum compensation
(4%) and flushed (9-9.5%) tofu samples, while, a
small decrease of 3% in oxygen levels was observed
in air packed tofu. In long term storage study,
oxygen concentration decreased for air packaged
tofu however, there was no certain increase for
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 Indian Food Industry Mag
Vol 36 No 3, May-Jun 2017
vacuum compensation and flushed packaged tofu.
For carbon dioxide concentration the decrease and
increase of all the packaging material became equal
at certain point of time (Campenhout et al 2013).
Microbiology and shelf life of tofu
Microorganisms require nutritious substrate
to grow which is readily provided by tofu.
Yersiniosis is an infectious disease caused by a
bacterium of genus Yersinia. The particular
bacterium responsible for yersiniosis in humans is
Yersinia enterocolitica. The outbreak of yersiniosis
was reported by Aulisio et al (1983) associated with
tofu consumption led to infections in 87 persons.
Other types of microorganisms associated with
spoilage of tofu are Clostridium botulinum,
Salmonella typhimurium and Staphylococcus
aureus (Kovats et al 1984).
In the experiment conducted by Han et al.
(2001) on commercial sufu, presence of
microorganisms such as Bacillus cereus (103-105
CFU/g), Clostridium perfringens (<10 3 CFU/g),
Staphylococcus aureus and Listeria monocytogenes
were reported in samples of sufu procured from
different markets of China and the Netherlands.
Based upon their color and flavor sufu is classified
into three types white, red and grey sufu in China.
Most of samples were high in mesophilic aerobic
bacteria (>105 CFU/g) out of which 85% were
gram-positive bacteria, lactic acid bacteria were
identified at a range of 105-107 CFU/g. White and
grey tofu samples had negative results for Listeria
monocytogenes, enterobacteriaceae and fungi,
hence it was concluded that commercial sufu is safe
to consume. Anbarasu and Vijayalakshmi (2007)
studied the effect of tulsi (Ocimum sanctum) extract
on the shelf-life of tofu at ambient temperature.
Tulsi extract elongated the shelf life of tofu, the
aerobic count of non-treated fresh tofu and tulsi
treated tofu samples had 5.96 x 102 and 2.37 x 103
CFU/g, respectively. Seven days of storage study
revealed total microbial count of 6.21 x 108 CFU/g
in non-treated tofu and 1.08 x 109 CFU/g of tulsi
treated samples. Yeasts and molds count in fresh
and tulsi treated tofu remained the same (<1.0 x
102), after 7 days. The tulsi extract treated tofu was
acceptable for more than 7 days without the use
TARs
of refrigeration.
Cloves have various essential oils which acts as
antibacterial agent and can be used for microbial
destruction in food. Cui et al (2015) studied the
20
effect of clove oil with liposome encapsulated on
the microbial activity of E. coli and S. aureus and
its application to tofu. Chemical stability of clove oil
is less to enhance the stability liposome
encapsulation was done. Clove oil treatment of
about 4 to 8 h, registered a reduction of E. coli and
S. aureus population to about 99.998%.
Modified atmosphere packaging helps in maintaining
the microbial count for tofu. Air packaged tofu had
higher microbial count (1-4 log cycles) compared to
the modified atmosphere packaged tofu and
inhibited the growth of microorganisms and kept
safe for consumption up to 14 days (Campenhout
et al 2013).
High hydrostatic pressure (HPP) processing
has also been studied for the inactivation of microbes
in tofu. HPP of food is influenced by several factors
such as type of microorganism, temperature of
processing, duration of high pressure treatment and
composition of the food. According to the factors
suitable pressure treatment is applied for the
inactivation of vegetative cells, pathogens and
spoilage causing microorganisms in food. Prestamo
et al (2000) studied the effectiveness of high
hydrostatic pressure treatment on tofu by applying
a pressure of 400 MPa at 5°C for varied
temperatures 5, 30 and 45 min. The treatments
showed decrease in microbial count from 5.54 x 104
CFU/g to 0.31, 1.56 and 2.38 log units with time
respectively. They reported reduction in
psycrotrophs and mesophiles by 2 log units and 1
log unit, respectively. In addition microorganisms
such as Salmonella, Pseudomonas and Gram-
negative bacteria were not detected post treatment.
Table 2 represents the probable microbial standards
of tofu, however no fixed microbial standards for
tofu has been reported by relevant bodies yet.
The storage life of tofu is relatively shorter
than its raw material, proving to be a major hurdle
for the commercial production of tofu on a large
scale. It has shelf life of near about a day in tropical
conditions. Park et al (2007) investigated effect of
turmeric (Curcuma aromatic Salab.) extract on the
shelf life of tofu at 25°C for 12 days. The bacterial
count and pH was found to be lower with
application of 0.02% turmeric extract. The bacterial
count was 1000 fold less in turmeric extracted tofu
than control tofu. It is postulated that spoilage of
tofu begins when the viable count crosses over 107
CFU/ml. Oyster shell has been studied in

association of tofu and 0.2% oyster shell
incorporation in tofu has reported to increase the
shelf life of tofu to upto 11 days at 10°C with an
acceptable level of viable microorganism (Kim et al
2007). Microorganisms responsible for spoilage of
tofu are represented in Table 2.
Conclusion
Tofu represents a product of soybean, mostly
consumed for its protein content. It is used in
various culinary dishes and health food for its
nutritious property and wholesomeness. Tofu is
largely preferred by the populations in eastern
countries such as Japan, Korea, Taiwan etc.
however, is gaining popularity all over the world.
Tofu is majorly manufactured by the unorganized
sector using traditional methods which leads to
unequal quality as there are no set standards of tofu.
Thus, a common international standards for tofu
needs to be established to promote
commercialization of tofu. Mechanization and
commercial scale production of tofu is practiced to
some extent, however, extensive study on making
efficient mechanized process for tofu production is
needed to fulfill the ever increasing demand of this
highly nutritious valuable foods product. The
perishability of tofu is a major constrain on
commercial scale production as the shelf life is 1-2
days at room temperature and around 11-12 days
in refrigerated conditions. Application of proper
packaging such as polyethylene, modified
atmospheric packaging (MAP) and use of natural
antimicrobials, have been found to reasonably
increase the practical storage life of tofu. Tofu can
prove to be our most efficient weapon against global
hunger and malnutrition problems provided
sufficient efforts are made to popularize its
nutritional significance amongst masses and efforts
are made to develop technologies for its industrial
production.
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