ELSEVIER

Characterization of three different

potato starches and kinetics of their
enzymatic hydrolysis by an cll-amylasc
T. Heitmann, E. Wenzig, and A. Mersmann

Department B of Chemical Engineering, Technical University of Munich,

Munich, Federal Republic of Germany

Three types of potato starch have been hydrolyzed by an a-amylase from Bacillus subtilis. The courses of
enzymatic degradation were observed with gel-permeation chromatography. The changes in molar mass distri-
butions by enzymatic attack alter (depending on the type of starch) the enzyme concentration and incubation time.
Additionally, macromolecules of native potato starch are destroyed by mechanical treatment and acid hydrolysis.
Initial distributions of molar mass for partly soluble and native potato starch are bimodalfiom the interpretation
of the clustered structure of amylopectin. The kinetics of the reaction are described with the Michaelis-Menten
rate equation with the specific activityfor native potato, partly soluble, and Zulkowski starch of 3,000; 2,900: and
2,630 umol mg-’ protein min-‘, respectively. The Michaelis-Menten parameters are 1.4, 1.9, and 6.0 g I-‘,
respectively. The significantly higher K, value of Zulkowski starch is explained by the low degree of polymer-
ization and the content of a-l,6 branching bonds. 0 1997 by Elsevier Science Inc.

Keywords: Starch; enzyme kinetics; alpha-amylase; molar mass distribution; mechanical degradation

Introduction perature of 60”C3 destroy crystallinity.4 Amylose is a poly-

The enzymatic hydrolysis of starch is widely investigated
by numerous scientists. The kinetics of starch degradation is
mostly measured by the velocity of liberation for reducing
ends. Starch is treated as a homogeneous substrate. In spite
of this, starch is one of the oldest “biotechnological” prod-
ucts from human history. Only a few facts about the mo-
lecular mechanism of enzymatic attack and degradation of
the starch molecules exists. The aim of this work is to
characterize native potato starch and partly soluble and
completely soluble potato starch solutions in order to de-
scribe the mechanism of their enzymatic cleavage; there-
fore, in this study the measurement of activity of starch-
degrading enzymes and the analysis of the hydrolyzed
starch types are linked. Native potato starch is a mixture of
77% amylopectin and 23% amylose.’ About 70% of the
starch granula is assumed to be amorphous. The crystallites
of amylopectin are built of alternating crystalline and amor-
phous zones.* Temperatures above the gelatinization tem-
Address reprint requests to Dr. -lng Alfons Mersmann, Prof., Technical
University of Munich, Arcisstrasse 21, 80333, Munich, Germany
Received 18 December 1995; accepted 8 May 1996
mer built of linear o-1,4-linked glucose monomers. Amy-
lopectin differs from amylose by o-1,6 branching of linear
amylose chains.
Two types of basic chains are known: type A and B with
a degree of polymerization (DP) of 15 and 45 glucose units,
respectively. Chains which are connected to the main body
are named Type B and those which are associated with the
B-type chains but not at the main body are named A type.5
Chemical or thermal treatment of starch leads to its deg-
radation. In this way, the other starch types used, namely
partly soluble and Zulkowski starch, are produced. They
differ in the degree of prehydrolysis. Partly soluble starch is
obtained by mild acid hydrolysis (Lintner method). Fully
soluble starch is produced at 190°C under the treatment of
glycerol (Zulkowski method, indicated by the manufac-
turer). Colonna et a1.6 showed that lintnerization degrades
all molecules simultaneously, but in articular, the amor-
phous regions of the starch granula. b is leads to higher
crystallinity of the fragments. On a molecular level, the acid
hydrolysis of polysaccharides (dextran) predominantly at-
tacks the o-1,4 linkages rather than the o--1,6 linkages.7
In the thermal process of extrusion-cooking, the o-1,4
linkages in amylose are hydrolyzed faster than the o-1,4
and a-l ,6 linkages in amylopectin.3 ‘Besides the thermal

Enzyme and Microbial Technology 20:259-267, 1997

0 1997 by Elsevier Science Inc. 0141-0229/97/$17.00
655 Avenue of the Americas, New York, NY 10010 PII SO141-0229(96)00121-4
Papers
depolymerization, redistribution of the chain linkages oc-
curs which forms highly branched molecules.3
On the contrary, milling preferentially degrades amylo-
pectin which can be explained by the bigger size of amy-
lopectin macromolecules which leads to a greater suscepti-
bility for rupturing linkages between clusters rather than the
smaller amylose molecules.4
This article deals with the influence of three starch types
differing in molar mass and internal linkages on the kinetics
of enzymatic hydrolysis with the enzyme o-amylase. This
enzyme from Bacillus subtilis is known to attack the cx-1,4
linkages of amylose in a random fashion.*
The enzymatic hydrolysis of starches by o-amylase
leads to the formation of smaller maltosaccharide frag-
ments. Every starch polysaccharide molecule possesses ex-
actly one reducing end which can reduce dinitrosalicylic
acid; therefore, the measurement of reducing sugars is a
useful method to determine the molar concentration of
starch molecules in solution.* The kinetics of liberation of
reducing sugars in the course of enzymatic starch degrada-
tion are described with Michaelis-Menten kinetics.9*‘0 An
overview is given by Komolprasert and Ofoli. l 1
The causes of different kinetic behavior are assumed to
be in the molecular properties of the starch types, hence the
initial and enzymatically hydrolyzed starch types are exam-
ined by gel-permeation chromatography (GPC), a rapid
method to measure molar mass and its distribution. This
method of analysis is reported by a number of research-
ers 59.12
Additionally, the enzyme iso-amylase was used to cleave
specifically the a-1,6 linkages in the starches (a method
used by Park and Rollings)9 to get further information about
the structure of internal linkages of the starch types.
Materials and methods
Enzymes
The enzyme o-amylase from B. subtilis was obtained from Fluka
(Neu-Ulm, FRG) and the iso-amylase from Pseudomonas umy-
loderumosu was obtained from Sigma (Deisenhofen, FRG).
Enzyme assays
Alpha-amylase activity was assayed by measuring the liberated
reducing sugars from 1% starch solution by the dinitrosalicylic
acid method (DNSA)13 at 60°C with acetate buffer at pH 6.
Samples (500 pl) were incubated with enzyme solution under
aerobic conditions for 10 min. After mixing with the DNSA re-
agent and heating at lOO”C, the absorbance was read at 530 nm
with maltose as standard. One unit (U) of enzyme activity is de-
fined as the amount of enzyme releasing 1 pmol of reducing
sugars min-I. The concentration of o-amylase in the samples was
between OS-25 mg 1-I with a protein content of 1.36% which was
measured by the method of Lowry et aLI using bovine albumin
(Fluka) as standard.
The rates were the means of at least three independent experi-
ments. The specific activity was determined by dividing the activ-
ity with the protein content of the sample.
For measurements of the course of hydrolysis, a 0.2-l fully
baffled reactor with 0.175-l working space was used. It was
equipped with a rushton turbine stirring at 800 rpm. The tempera-
ture was held constant at 60°C. The reactor was filled with 100 ml
260 Enzyme Microb. Technol., 1997, vol. 20, March
of 1% starch solution. After the temperature was adjusted, 10 ml of
o-amylase solution was injected and samples were taken every
hour.
Starch degradation with iso-amylase was conducted in 500 u-1
samples incubated with enzyme solution under aerobic conditions
at 40°C with acetate buffer at pH 3.5. The concentration of iso-
amylase in the samples was 1.6 mg protein I-’ [calculated with the
added volume and the protein content per volume (0.8 mg ml-‘)
given by the manufacturer]. The iso-amylase did not degrade amy-
lose.
Mechanical degradation
A sample of autoclaved 1% starch solution was stirred at very high
shear rates in an Ultra-Turrax stirrer (T25, IKA-Labortechnik,
Janke & Kunkel, FRG) at 20,000 rpm for one min three times and
were analyzed afterwards via GPC. Sample warm up was moder-
ate.
Acid hydrolysis
To 100 ml of 1% autoclaved native potato starch solution, 1 ml of
37% HCl (Fluka) is added which results in a concentration of
0.119 N HCI. The solution is heated to 60°C. Samples are taken
every hour.
Analytical methods
Molar mass distribution and maltose content were measured by
gel-permeation chromatography (OHpak KB-803 column and
OHpak KB-805 column, Shodex, Japan) at 20°C with 1 ml mini
0.05 M NaNO, eluent flow.
Chemicals
Partly soluble starch was obtained from Sigma (Deisenhofen,
FRG), Zulkowski starch from Fluka, and native potato starch from
Brtiggen (Liibeck, FRG). Only Zulkowski starch is soluble in cold
water without any detergents. The partly soluble and especially
native potato starch are mainly insoluble at normal conditions in
water. By fit cooking and afterwards autoclaving at 12 1“C for 20
min clear homogeneous solutions in higher concentrations (up to
40 g 1-i potato starch) were obtained. After cooling, only the
viscosity of native potato starch solution increased (opposite to the
initial suspension) and the flow behavior was pseudoplastic. Fresh
solutions were always used. The samples were stored under sterile
conditions after autoclaving.
Results
Initial starch molar mass distributions
To explain the results, the starches have to be investigated
in more detail. For determining the initial molar mass dis-
tribution of the different starches, GPC analysis was used.
Figure 1 shows the initial logarithmic molar mass distribu-
tions of the starches versus the DP calculated with Eqs. (1)
and (2):
(1)
and
1 dm loge
w,,,(M)=*-=--
mges d(ln M) mges d(log M)
(2)
 Characterization of three different potato starches: T. Heitmann et al.

,,

‘\
\
\
\
\
\
,’
/!L \
/ \
\
\
\

L b
lo: (1’
Figure 1 Initial starah molar mass distri-
degree of polymerisation DP [-] - butions

The average molar masses are numerically calculated by the
moments of molar mass:
and
(4)
The calculated average molar masses of the three starch
types are listed in Table 1.
Both the partly soluble and native potato starch show
bimodal molar mass distributions. The peak of higher molar
mass of native potato starch is exactly in the range given for
amylopectin of 107-lo9 g mol-I.*
The enzyme iso-amylase was used to cleave specifically
the cl--l,6 linkages to get more information about the inter-
nal linkages of the starches. The results of hydrolysis after
sufficient time (no further change in distribution was ob-
served) are shown in Fzjpre 2.
Starch degradation with this enzyme leads to a drastic
drop in the molar mass of the partly soluble and native
potato starch (degradation of Zulkowski starch was not de-
tectable). A small fraction with higher molar mass of both
Table 1 Kinetic properties of starches depicted in Figure 1 and
Figure2
starches resists the enzymatic attack. These fractions con-
tain approximately 20% (w/w) of the total mass of both
starches.
The fractions of lower molar mass liberated by iso-
amylase are nearly equal for both starches and show distri-
butions of A- and B-chain types with a DP of 17 and 51,
respectively. These fractions are in the flame range as initial
Zulkowski starch.
h4echanical disruption of potato sttirch
The polysaccharides of the fraction of higher molecular
mass are degraded by stirring. As seen jn Figure 3, a part of
the second peak representing the fractjon of higher molar
mass decreased after every stirring se&on even in the first
min (initial distribution not shown in $igure 3; see Figure
1). First, the mechanical disruption dihinishes the fraction
of higher molar mass due to the attack of the biggest mol-
ecules. After three min, the peak nearlp disappeared.
On the contrary, it was not possiblei to erase the second
peak of partly soluble starch under the same conditions.
Changes of molar mass distribution in the course of
acid hydrolysis
Similar to the mechanical disruption first, the fraction of
higher molar mass of native potato sta$ch is degraded (Fig-
ure 4). The products are uniformly didtributed in the range
of the lower molar mass fraction. With further degradation,
this fraction splits in a bimodal distribution similar to the
distribution of initial partly soluble starch (Figure 1).

Specific
activity a Michaelis-Menten
Changes of molar mass distributidn in the course of
(pm01 mg-‘) K,,,(g I-') enzymatic hydrolysis
With the used c+amylase concentratjon, the native potato
Zulkowski starch 2,630 6.0
Partly soluble starch 2,900 1.9 starch is degraded quickly (1 h) to saccharides (Figure 5)
Native potato starch 3,000 1.4 which shows a distribution similar tci initial partly soluble
starch (Figure 1). With further degradation, the fraction of

Enzyme Microb. Technol., 1997, vol. 20, March 261

Papers

0.8

1 ---- native starch

0.6

4 a
O3 07
Figure 2 The starch types after 8 h of
degree of polymerisation DP [-] incubation with the enzyme iso-amylase

higher molar mass completely disappeared. This was ac-
companied by the production of oligosaccharides over the
entire smaller range which was not uniformly distributed
but demonstrated a preference for higher molar mass and
especially maltosaccharides with a DP of 8. Using a lower
enzyme concentration in the first stages of hydrolysis of
native potato starch was investigated in more detail. The
disappearance of the second peak (mentioned before) is
again visible in F@gure 6. With further hydrolysis, the
“movement” of the former first peak can be observed.
Also, the appearance of a new peak with a DP at approxi-
mately 180 is noted. After 9 h, the distribution is equal to
that of F&M-~ 5 after 1 h.
The course of hydrolysis of partly soluble starch was also
examined, but the data does not need to be shown because
the initial molar mass distribution was similar to that of
native potato starch in Figure 5 after the first hour. The
further changes in distribution are also approximately simi-
lar to those of native potato starch in that Figure.
The last starch investigated was the Zulkowski starch;
the course of hydrolysis is shown in Figure 7. This type of
starch is degraded slower since it is expected from the mea-
surements of reducing sugars presented above. After long
incubation periods, the hydrolysis of Zulkowski starch is
visible by slightly altered distribution. Besides the liberation
of maltosaccharides with DP 8, the formation of maltose
occurs. The changes are similar to the last stages of hydro-
lysis depicted in Figure 5.
Enzyme kinetics
The substrate of the enzyme o-amylase are the o-l ,4 bonds
in the polysaccharide molecule. The concentration of these

5
‘S
$

4
-0
ul
UI -
EL.
0.8 -

0.6 -
7-r 3fLin
\
,’
-.
\
‘,
1 min

5 B
2 3 0.4 -
.9
E
Jz /
.z
ca 0.2 -
.._
$

L LL
IO! l( 1’ Figure3 Course of degradation of native
potato starch during treatment with high
degree of polymerisation DP [-] - stirring intensity in 1 min steps

262 Enzyme Microb. Technol., 1997, vol. 20, March

 Characterization of three different potato starches: T. Heitmann et al.

initial native starch

Figure 4 Course of acid hydrolysis of na-

degree of polymerisation DP [-] - tive potato starch (Sk, 0.119 N HCI)

linkages is approximately proportional to the polysaccha-
ride mass concentration. The deviation due to the o-1,6
bonds in potato starch is small if the concentration of those
in the polymer is low. Therefore, Michaelis-Menten kinetics
are used to describe the velocity of liberation of reducing
sugars v as a function of the initial starch concentration S:
a cE S
(5)
v=K,
The rate is proportional to the enzyme (protein) concentra-
tion cn with a specific activity a and the Michaelis-Menten
parameter K,.
The rates of hydrolysis of all three starch types are de-
scribable with Eq. (5). The measured and calculated rates
are depicted in Figures 8 and 9 with Lineweaver-Burk plots.
The values of the specific activity a and the Michaelis-
Menten parameter K, for native potato, partly soluble, and
Zulkowski starch are listed in Table 2.
As a result at low substrate concentrations, the affinity of
the enzyme o-amylase for the substrate Zulkowski starch
was lower compared to the other starches. This was re-
flected by the higher K, parameter. The rates of hydrolysis
of native potato and partly soluble starch are similar. Partly
soluble starch shows a slightly higher Krh parameter in com-
parison with native potato starch. The rate of enzymatic
starch degradation is not significantly iinhibited by the ad-
dition of maltose (data not shown).
Discussion
Characterization of the starch molecules
Autoclaved native potato and partly soluble starch solutions
are bimodal distributed. What are the causes for these dis-

I I
--- 1 hour
--- 2 hours
---- 4 hours
- 9 hours

$ initial native’ starch

g 3 0.4 \
.o
sE ,-.. .
3 !'
0.2
/I/--
B
_*'y,_
-_A ,
__A
3 0 I! II II Figure 5 Course qif enzymatic hydrolysis
IO0 10’ ICt7 of native potato1 starch (0.45 mg 1-l
degree of polymerisation DP [-] - a-amylase)

Enzyme Microb. Technol., 1997, vol. 20, March 263

Papers
, of native potato starch with a lower en-
s IO'
tributions? They cannot be explained by the approximately
20% amylose content of native potato starch because the
o--1,6 specific hydrolysis with the enzyme iso-amylase de-
graded the initial distributions except for a small resisting
fraction of higher molar mass (formerly overlayed by the
other fractions) which agrees in molar mass and the percen-
tual content with the given data of amylose at 20% and
2.6-6.5 * IO5 g mol-1,2 respectively. The amylose from
partly soluble starch has lower molar mass than that of
native soluble starch due to lintnerization.
The bimodal distribution of autoclaved native potato
starch can be removed by mechanical degradation. The sec-
ond peak of native potato starch results from the cleavage of
the linear chain portions of amylopectin by high stirring
intensities (Figure ZOb). The broad but not uniform distri-
bution of the increasing fraction indicates that there are
linkages of the amylopectin molecule being favorly attacked
r r
initial Zulkowski starch
, , ,,, , , , , ,,,,
lyFLisatioA”AP
264 Enzyme Microb. Technol., 1997, vol. 20, March
--- 1 hour
--- 2 hour
---- 3 hour
- 5 hour
initial natibe starch
II
Figure 6 Course of enzymatic hydrolysis
106 10'
zyme concentration (0.14 mg I-’ cu-amy-
[ - jl”“-
lase)
and an upper size limit for mechanical degradation. Analo-
gously, the smaller molar fraction of clusters is ruptured
from the big amylopectin molecules of native potato starch
(Figure IOU) by the thermal degradation during autoclaving.
During the first stages of acid and enzymatic hydrolysis
of native potato starch, the fraction of higher molar mass is
also preferentially attacked (Figure 6). Predominantly, the
external o-1,4 linkages outside the cluster regions rather
than the internal bonds are cleaved. These results cannot be
explained with the model proposed by Park and Rollingsg
about inhibition of enzymatic attack by steric effects of the
regional network of concentrated polysaccharide chains in
the cluster domain of amylopectin due to similar results of
acid hydrolysis. This effect must be related to different re-
activities of the longer chains connecting the clusters com-
pared to the internal bonds of the clusters.
With further enzymatic degradation, a bimodal distribu-
, , , , ,,(,
Figure 7 Course of enzymatic hydrolysis
of Zulkowski starch (0.45 mg I-’ a-amy-
[ - ;‘- lase)
 Characterization of three different potato starches: T. Heitmann et al.

Figure 8 Hydrolysis rate of different

0 starch types versus concentration with
the enzyme a-amylaie. The lines denote
starch concentration S [ g / I] the calculated rates

tion appears again (Figure 5) which is nearly identical to
that of partly soluble starch. This type of starch is industri-
ally produced by acid hydrolysis according to the Lintner
method. The bimodal distribution of autoclaved lintnerized
potato starch is proposed as being composed of clusters
(Figure 10~) and small fragments (Figure 106).
With further enzymatic attack, the cluster structure dis-
appears and the formation of maltosaccharides increases.
Kinetics of starch degradation
The kinetics of enzymatic hydrolysis of different starch
types are described with the velocity of liberation of reduc-
ing sugars. The results of the kinetics demonstrate that the
maximal velocity of the liberation of reducing sugars from
the investigated starch types decreases while the K,,, value
increases with decreasing molar weight of the starch types.
Similar results are reported by Fujii and Kawamura” for the
same enzyme used here that acts on starch. They measured
constant maximal velocities and increasing Km values with
decreasing molar mass. The activity at substrates with a
molar mass lower than 5,000 g mol-’ was reported to be
negligible.
The first attempt to explain the increase in the Km pa-
rameter and the decrease in the maximal velocity of the
Michaelis-Menten kinetics with decreasing molar mass was
a noncompetitive inhibition by a substaqce which is part of
the substrate. In the literature, it was reported that maltose
noncompetitively inhibits starch hydqolysis by a-amy-
lase.15 but our experiments show that nb significant inhibi-

I I Figure 9 Lineweayer-Burk plot of hydro-

1 2 lysis of the differem starch types with the
enzyme a-amylasel The calculated values
reciprocal starch concentration 1 / S [ I / g ] are plotted with lines

Enzyme Microb. Technol., 1997, vol. 20, March 265

Papers

Michaelis-Menten parameter must be sought in the molar

Table 2 Molar mass averages of starch types depicted in Fig- size and influence of o-l,6 bonds. In this context, the theory
ure 3 of subsites should be mentioned which suggests that sub-
strates with glycosidic linkages in the range or less than the
Number average Weight average number of enzyme subsites are hydrolyzed slower.‘“,r7 Col-
molar mass m” molar mass Efw onna et aL4 gives a value of ten monomers before the re-
(g mol-‘) (g mol-‘1
action rate decreases. These conditions are given for
Zulkowski starch which has a small number average molar
Zulkowski starch 1,900 7,500 mass (DP 12). From the chromatograms of initial
Partly soluble starch 20,000 1.4 x 105
Native potato starch 84,000 3.7 x IO’
Zulkowski starch, it has been calculated that approximately
20% (w/v) have a DP less than 9. Additionally, the a-1,6
linkages in the Zulkowski starch molecule lead to a further
reduction in the available bonds by three causes:
1. They are not hydrolyzible for a-amylase
tion by maltose occurs. Another possibility would be the 2. They divide the molecule in linear portions with an av-
inhibition of limit dextrins, but they only effect competitive erage degree of polymerization much lower than the ini-
inhibition.9 A further explanation for the four-time higher tial one
K,,, value of amylose compared to amylopectin is given by 3. In the neighborhood of a-1,6 linkages, the a-1,4 link-
Park and Rollings:9 The stiff helical structure of amylose in ages are uncleavable for the enzyme. Namely, three
a dimethyl sulfoxid-water system may be difficult for the cx-1,4 bonds per a-1,6 linkages are reported.9
enzyme to grasp, but in the pure water system used here at
60°C the starch fragments give no blue color with iodine. To calculate the number of a-1,6 bonds per molecule
They must, therefore, be assumed as oriented in a random B 1,6r the total number of bonds B must be known:
coil order.
The causes for variation in the specific activity and the B=DP-1 (6)

With the molar mass of glucose in a polysaccharide chain of

amylopectin molecule 165 g mol-’ and the number average molar mass (Table 2),
- Eq. (6) gives a total number of bonds per molecule of B
= 508 for native potato starch and B = 11 for Zulkowski
starch. By dividing this number of bonds minus one, it gives
the number of bonds (approximately 43) that must be
cleaved to obtain Zulkowski starch from native starch.
Amylopectin contains approximately 5% of a-l,6 bonds.18
Incorporating the native potato starch which contains 23%
amylose in starch, 3.85% of the linkages are o-1,6 bonds.
With the assumption that only the a-1,4 bonds are cleaved
during acid hydrolysis, the part of a--1,6 bonds increases to
4.20%. For Zulkowski starch, this results in less than one
a-1,6 lmkage per molecule (B,,, = 0.46). This value must
be too low, which brings to mind the results of o-1,6 spe-
cific iso-amylase which does not considerably attack
Zulkowski starch. This means that either there are no o-l,6
amylopectin fragment bonds in Zulkowski starch or that they are repeated too
close in the chain (with only a few a-1,4 linkages between
them) for being hydrolyzed by this enzyme because iso-
amylase is not able to degrade pullulan; hence, there are
only three a-1,4 linkages between the a-1,6 bonds for this
substrate.
The extra a--1,6 linkages in the Zulkowski starch mol-
partly soluble starch ecule may originate from the thermal treatment by the in-
dustrial production which leads via transglycosylation2 to
further a-1,6 linkages; however, further investigations are
necessary to obtain more information about the structure of
Zulkowski starch in detail.

d) Zulkowski starch
Acknowledgments

We thank the Max Buchner-Forschungsstiftung (MBFSt-

Figure 10 Model for amylopectin degradation Nr. 1749) for financial support of this work.

266 Enzyme Microb. Technol., 1997, vol. 20, March

 Characterization of three different potato starches: T. Heitmann et al.

List of symbols liquefaction: Simulation of the high-molecular-weight product dis-

tribution. Biotechnol. Eioeng. 1984, 26, 1475-1484
pm01 min-’ mg- ‘; Specific activity 6. Colonna, P., BulBon, A., and Lemarie, F. Action of Bacillus subtilis
o-amylase on native wheat starch. Biotechnol. Bioeng. 1987, 31,
-; Total number of bonds per molecule
895-904
B 1.6 -; Number of o-1,6 bonds per molecule 7. Jeremic, K., Ilic, L., and Jovanocic, S. Kinetics of dextran depoly-
CE mg 1-l; Enzyme (protein) concentration merisation in aqueous HCl. Eur. Polym. J. 1985, 21, 537-540
DP -; Degree of polymerization 8. Hiromi, K., Ogawa, K., Nakanishi, N., and Ono, S. A kinetic
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Kll of linear high polymer by using an exo-enzyme. J. Biochem. 1966,
M g mol-‘; Molar mass
69,439-449
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G” g mol-‘; Number average molar mass acteristics on kinetics of enzymatic depolymetization of mixed lin-
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mDP g; Mass of fraction DP glucoamylase on hydrolysis of starch. Biotechnol. Bioeng. 1985.27,
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51, 209-223
ages mol 1-r; Total amount of moles
12. Henriksnas, H. and Liivgren, T. Chain-length distribution of starch
P g 1-l ; Product concentration hydrolyzate after o- or B-amylase action. Biorechnol. Bioeng. 1978,
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