The Journal of The Textile Institute, 2014

Vol. 105, No. 2, 203–208, http://dx.doi.org/10.1080/00405000.2013.834574

Reaction kinetics study of α-amylase in the hydrolysis of starch size on cotton fabrics
Kang-li Fu and Da-nian Lu*

Key Laboratory of Science & Technology of Eco-Textile, Ministry of Education, College of Chemistry, Chemical Engineering &
Biotechnology, Donghua University, Shanghai, China
(Received 29 May 2013; accepted 9 August 2013)

The hydrolysis reaction kinetics of α-amylase on the starch size of cotton fabrics followed the first-order reaction in the
initial stage that α-amylase hydrolyzed on the surface of cotton fabrics; desizing ratios of more than 95% were obtained
in the initial stage, which are acceptable, and the hydrolysis time for the initial stage shortened sharply as the
temperature increased. The increase of α-amylase dosage enhanced the reaction rate constant as well as speeded up the
first-order reaction rate in the initial stage, both treated at the same treatment temperature. The hydrolysis action of
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α-amylase on starch size on the surface of cotton fabrics speeded up significantly as the temperature increased. Also, the
treatment time needed to obtain an acceptable desizing ratio (> 95%) shortened sharply from 6 to 1/12 h as the treatment
temperature increased from 20 to 100 °C.
Keywords: α-amylase; cotton fabrics; desizing; kinetics

Introduction Meyer, Proth, Strehaiano, & Pingaud, 2003; Naveena,

Cotton warp yarns have to be sized prior to weaving to
apply a protective coating to improve yarn strength and
reduce yarn hairiness. Starch-based products are most
frequently used for slashing cotton yarns (Cai, Qiu,
Zhang, Hwang, & Mccord, 2003; Li & Qiu, 2012).
There are sizing agents distributed both on the surface of
cotton fabrics and in the interior of cotton fibers after the
sizing process (Shen, Xue, Zhang, Liu, Gao, & Cui,
2011; William & Yelland, 1939; Yang & Bresee, 1987).
Removal of the starch size after weaving is important to
ensure optimum results in the subsequent processes, in
which the fabrics is generally scoured, bleached, and
dyed (Eren, Anis, & Davulcu, 2009; Peng et al., 2010).
In the past, sodium hydroxide of high concentration
was used as a desizing reagent, which produced great
effluents and damaged the environment seriously
(Buschle-Diller, Yang, & Yamamoto, 2001; Csiszár
et al., 2001). More recently, α-amylase has been
introduced to the hydrolysis of starch size of cotton
fabrics. This method produces little effluent and is
friendly to the environment (Tzanov, Calafell, Guebitz,
& Cavao-Paulo, 2001; Vigneswaran, 2012). Some
reports showed the kinetics hydrolysis reaction of
α-amylase on starch in solution following the first-order
reaction (Al-Rabadi, Gilbert, & Gidley, 2009; Ezeogu,
Duodu, & Taylor, 2005; Marc, Engasser, Moll, &
Flayeux, 1983) or the second-order reaction (Brandam,
Altaf, Bhadrayya, Madhavendra, & Reddy, 2005). But,
there are no reports on the kinetics of hydrolysis reac-
tion of α-amylase on the starch size of cotton fabrics.
Because starch size is distributed both in the interior of
cotton fibers and on the surface of cotton fabrics (Shen
et al., 2011; William & Yelland, 1939; Yang & Bresee,
1987), there was a big difference between the hydroly-
sis action of α-amylase on starch size of cotton fabrics
and on starch in solution.
Generally, the process of desizing is performed by
the means of cold pad batch treated at room temperature
for several hours with mesophilic α-amylases (Csiszar &
Fekete, 2010; Hashem, EI-Bisi, Sharaf, & Refaie, 2010);
the disadvantage of this method is the long enzymatic
treatment time. Moreover, thermophilic α-amylase that
can be used with a wide temperature range of 20–100 °C
are introduced into the biodesizing process; desizing with
this thermophilic α-amylase reagent can be conducted at
95–100 °C with very short enzymatic treatment time
(Dong & Lu, 2008).
In this study, desizing of cotton fabrics was
conducted at room temperature of 20 °C (represented
desizing in winter) and 35 °C (represented desizing in
summer) and the kinetics of hydrolysis reaction
mechanism were discussed. Moreover, the hydrolysis
action of thermophilic α-amylase on the starch size on
the surface of cotton fabrics was studied with a tempera-

*Corresponding author. Email: danianlu@163.com

! 2013 The Textile Institute

 204 K.-l. Fu and D.-n. Lu
ture range of 20–100 °C and the desizing treatment times
were compared.
Materials and methods
Materials
α-Amylase (Suhong desizyme 2000L) used in this study
was obtained from Novozymes A/S, starch-sized cotton
fabrics (plain-woven 100%, 120 g/m2), supplied by a
Shenda, the second printing and dyeing factory
(Shanghai, China). Nonionic surfactant Java Foundation
Classes was supported by Transfar Group Co. Ltd,
(Zhejiang, China). Other reagents used in this work were
of analytical grade and used as received without further
purification. The buffer solution used in this article was
acetic acid–sodium acetate (pH 6.5), which was prepared
by 0.2 M acetic acid and 0.3 M sodium acetate.
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Desizing process of cotton fabrics
Desizing process of starch-sized cotton fabrics was treated
with α-amylase by pad batch at temperature range of 20–
100 °C. Starch-sized cotton fabrics were dipped into the
bath (pH 6.5) containing X g/L α-amylase and 1 g/L non-
ionic surfactant Java Foundation Classes. Dipping and
padding was done twice with wet pick up of 100%. The
starch-sized cotton fabrics with pick up of 100% were
dwelled at 20–100 °C for a certain period of time, and then
the cotton fabrics after enzymatic treatment were washed
thoroughly with boiling water and cold water separately;
then, the desized cotton fabrics obtained were dried.
Preparation of starch standard curve
Starch was dissolved in 50 mL distilled water and 15 mL
of 42% perchloric acid were put into it with constant
stirring until the starch dissolved completely; then the
mixture was neutralized with sodium hydroxide. A cer-
tain volume of this starch solution was blended with
solution of iodine and potassium iodide. The mixture
were kept away from light for 5 min to complete the
color reaction and the absorbance at 620 nm was
detected in a Unico UV-4802 spectrophotometer.
Determination of the desizing ratio
The starch size content on starch-sized cotton fabrics,
before and after enzymatic desizing treatment, was deter-
mined for the calculation of desizing ratio. The content
of starch size on starch-sized cotton fabrics was analyzed
by referring to perchloric acid method (Rose et al.,
1991). The starch-sized cotton fabrics were washed and
dried at the end of the enzymatic treatment. Then, they
were cut into small pieces and put into 30 mL of 42%
perchloric acid with constant stirring for 10 min at room
temperature. The mixture was neutralized with sodium
hydroxide, followed by filtration.
The concentration of starch size in filtrate was
detected according to the starch standard curve. Desizing
ratio (D%) of starch-sized cotton fabrics was defined by
the following equation:
W0 " W1
D% ¼ # 100;
W0
where W0 (mg/g cotton fabrics) is the starch weight on
the fabrics before desizing and W1 (mg/g cotton fabrics)
is the starch weight on the fabrics after desizing.
Results and discussion
Effect of α-amylase dosage and batching up time on
desizing ratio of fabrics
Figures 1 and 2 show that the desizing of starch-sized
cotton fabrics get enhanced as α-amylase dosages
increase. While there are no obvious changes in the
desizing ratio of starch-sized cotton fabrics after
α-amylase dosages exceed 0.10% (owf) in both these
two figures, the α-amylase dosage with 0.10% (owf) is
the optimum dosage of enzyme applied. Desizing ratios
of starch-sized cotton fabrics cannot achieve 100% even
after treatment at 35 °C for 10 h with α-amylase dosage
of 0.30% (owf). Moreover, desizing ratios of starch-sized
cotton fabrics are improved by prolonging the batching
up time; there are no significant changes after batching
up time comes to 6 and 4 h (Figures 1 and 2) separately.
Nearly same desizing ratios of starch-sized cotton fabrics
treated by the same α-amylase dosages are acquired at
20 and 35 °C; for example, a desizing ratio of about
46% is obtained by enzymatic treatment with α-amylase
dosage of 0.025% (owf) both at 20 and 35 °C.
Figure 1. Effect of batching up time on desizing ratio of
starch-sized cotton fabrics with different α-amylase dosages
treated at 20 °C.
 The Journal of The Textile Institute
Figure 2. Effect of α-amylase dosage and batching up time on
desizing ratio of cotton fabrics treated at 35 °C.
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Reasons of these phenomenon are shown as follows:
firstly, α-amylase on starch-sized cotton fabrics with wet
pick up of 100% by the cold pad batch was in a
relatively fixed position and the low moisture condition
was not good for its movement in large scale (Dianawati
& Shah, 2011), which implies that α-amylase catalyzed
the hydrolysis of starch size in the place around where
α-amylase was padded to. After being treated with
0.025% (owf) α-amylase by cold pad batch process, a
large area of starch-sized cotton fabrics could not come
into contact with α-amylase and most of the starch size
in this place was retained. Secondly, starch size on
cotton fabrics was saturated by α-amylase after the
dosage of α-amylase was increased to 0.10% (owf) and
almost all the starch size on the surface of cotton fabrics
was removed, but still a little starch size stayed in the
interior of cotton fibers which was not hydrolyzed by
α-amylase (Shen et al., 2011; William & Yelland, 1939;
Yang & Bresee, 1987). Thirdly, according to the
Arrhenius equation, the elevation of enzymatic treatment
temperature accelerates the catalytic rate of α-amylase;
so, compared to starch-sized cotton fabrics batched at
20 °C, starch-sized cotton fabrics batched at 35 °C with
the same α-amylase dosage shortened the batching up
time to some extent.
Reaction kinetics study of α-amylase on starch size of
cotton fabrics
lnðCA Þ ¼ "kt þ lnðCA 0Þ; ð1Þ
kt ¼ " ln½ð1 " DS Þ=ð1 " DS0 Þ(; ð2Þ
In these two formulas, k represents the reaction rate
constant, t represents the reaction time, CA represents the
205
residual content of starch size of cotton fabrics after
enzymatic desizing treatment, CA0 represents the content
of starch size of cotton fabrics before enzymatic desizing
treatment, Ds represents the desizing ratio of cotton fab-
rics after enzymatic desizing treatment for t hours,
(1 – Ds) represents the residue ratio of starch size of
cotton fabrics after enzymatic desizing treatment for t
hours, and it was used instead of CA, Ds0 represents the
desizing ratio of cotton fabrics at the beginning of the
enzymatic desizing experiment, and (1 – Ds0) represents
the residue ratio of starch size of cotton fabrics at the
beginning of the enzymatic desizing experiment and it
was used to replace CA0.
Some reports showed that the hydrolysis reaction
kinetics of α-amylase on starch in solution followed the
first-order reaction (Al-Rabadi et al., 2009; Ezeogu et al.,
2005; Marc et al., 1983) or the second-order reaction
(Brandam et al., 2003; Naveena et al., 2005). There was
a big difference between the state of starch size of cotton
fabrics and that in solution; so, the hydrolysis action of
α-amylase on starch size of cotton fabrics is discussed.
Figure 3 shows there is a nice linear relation between
‘ln[(1 – Ds)/(1 – DS0)]’ and ‘batching up time’ with a
certain batching up time range of 0–6 h and 0–4 h at 20
and 35 °C, respectively. The linear relation between ‘ln
[(1 – Ds)/(1 – DS0)]’ and ‘batching up time’ turns to reces-
sion curve relation after the batching up time exceeds 6
and 4 h by batching at 20 or 35 °C, respectively. The
reason is attributed to the distribution of starch size in
cotton fabrics on the surface of cotton fabrics and in
the interior of cotton fibers after the sizing process
(Shen et al., 2011; William & Yelland, 1939; Yang &
Bresee, 1987). α-amylase hydrolyzed starch size on the
surface of cotton fabrics at the initial stage of
the cold pad batch and this hydrolysis process followed
the first-order reaction. α-amylase penetrated into the
Figure 3. Hydrolysis reaction kinetics of starch size of cotton
fabrics with α-amylase treated at 20 and 35 °C, respectively.
Note: α-amylase dosage used here was 0.025% (owf).
 206 K.-l. Fu and D.-n. Lu

order reaction, and these desizing ratios are acceptable

Figure 4. Hydrolysis reaction kinetics of starch size on the
surface of cotton fabrics with different α-amylase dosages
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(Peng, Gao, Sun, Yao, & Qiu, 2009); so, all the discus-
sion mentioned below are centered on the initial stage.
Both Figures 4 and 5 show hydrolysis reaction kinet-
ics of starch size on the surface of cotton fabrics with
different α-amylase dosages treated at 20 or 35 °C fol-
lowing the first-order reaction. Slope of every straight
line represents the hydrolysis reaction rate constant.
Table 1 shows that increasing of α-amylase dosage
enhances the hydrolysis reaction rate constant as well as
speeds up the first-order hydrolysis reaction rate in the
initial stage treated at both 20 and 35 °C. In the same
condition, groups treated at 35 °C show much greater
first-order hydrolysis reaction rate constants, which
implies that α-amylase treated at 35 °C displayed faster
hydrolysis action than that treated at 20 °C and this result
coincided well with conclusions of Figures 1 and 2.

treated at 20 °C.
Activation energy of α-amylase in the hydrolysis of
starch size on the surface of cotton fabrics

k ¼ Aeð"Ea =RT Þ ; ð3Þ

! "
k2 Ea 1 1
ln ¼ " " ; ð4Þ
k1 R T2 T1

Activation energy (Ea) of α-amylase in the hydrolysis

Figure 5. Hydrolysis reaction kinetics of starch size on the
surface of cotton fabrics with different α-amylase dosages
treated at 35 °C.
fiber vacuity to hydrolyze starch size in the interior of
cotton fibers as the gradually hydrolysis of starch size on
the surface of cotton fabrics and this process decreased
the hydrolysis rate, thus making the hydrolysis process
of starch size with α-amylase deviate from the first-order
reaction greatly. Desizing ratios of more than 95% were
obtained in the initial stage which followed the first-
of starch size on the surface of cotton fabrics was
calculated according to Equation (4) and it was 20.99 kJ/
mol. According to Equation (3), hydrolysis reaction rate
constants increased as the enzymatic treatment tempera-
ture increased, which means the hydrolysis action of
α-amylase on starch size on the surface of cotton fabrics
speeded up significantly and the enzymatic treatment
time shortened sharply as the temperature increased.
Desizing with α-amylase treated at different
temperatures
Desizing of α-amylase treated at different temperatures
was studied below. Enzymatic desizing in other
temperatures followed the first-order reaction during the
hydrolysis of starch size on the surface of cotton fabrics;
the hydrolysis time for the initial stage shortened sharply
as the the temperature increased and the data are not
shown here.

Table 1. First-order hydrolysis reaction rate constant of the hydrolysis reaction of starch size with different α-amylase dosages at 20
or 35 °C.

α-Amylase dosage (owf)

T (°C) 0.025% 0.05% 0.10% 0.20% 0.30%
20 0.0408 0.0811 0.3401 0.3425 0.3529
35 0.0602 0.132 0.4818 0.5227 0.5531
 The Journal of The Textile Institute
Table 2. Hydrolysis reaction rate constant and enzymatic treatment time needed to acquire desizing ratio of more than 95% at
different treatment temperatures.
20 35
"1
k (h ) 0.3401 0.4818
t (h) 6 4
Note: α-amylase dosage used here was 0.10% (owf).
Table 2 shows that hydrolysis reaction rate constantly
enlarged seriously from 0.3401 to 24.580 h"1 and the
enzymatic treatment time needed to obtain acceptable
desizing ratio (> 95%) shortened sharply from 6 to
1/12 h as the treatment temperature increased from 20 to
100 °C, which verifies the conclusion mentioned above.
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Conclusion
The kinetics hydrolysis reaction of α-amylase on starch
size of cotton fabrics followed the first-order reaction in
the initial stage that α-amylase hydrolyzed on the surface
of cotton fabrics; acceptable desizing ratios of more than
95% were obtained in the initial stage treated at 20 or
35 °C consuming 6 and 4 h, respectively. The increase of
α-amylase dosage enhanced the reaction rate constant as
well as speeded up the first-order reaction rate in the ini-
tial stage treated at both 20 and 35 °C. Groups treated at
35 °C showed much greater first-order reaction rate con-
stants than groups treated at 20 °C in the same condition.
The hydrolysis action of α-amylase on starch size on the
surface of cotton fabrics speeded up significantly as the
temperature increased and the treatment time needed to
obtain an acceptable desizing ratio (> 95%) shortened
sharply from 6 to 1/12 h as the treatment temperature
increased from 20 to 100 °C.
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