Optimization of rice wine fermentation process based on the simultaneous
saccharification and fermentation kinetic model

Dengfeng Liu, Hongtao Zhang, Zhizhong Lian, Baoguo Xu

PII: S1004-9541(16)30511-0
DOI: doi: 10.1016/j.cjche.2016.05.037
Reference: CJCHE 590

To appear in:

Received date: 16 October 2015

Revised date: 17 February 2016
Accepted date: 26 March 2016

Please cite this article as: Dengfeng Liu, Hongtao Zhang, Zhizhong Lian, Baoguo Xu,
Optimization of rice wine fermentation process based on the simultaneous saccharification
and fermentation kinetic model, (2016), doi: 10.1016/j.cjche.2016.05.037

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 ACCEPTED MANUSCRIPT

2015-0514

Process Systems Engineering and Process Safety

Optimization of rice wine fermentation process based on the

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simultaneous saccharification and fermentation kinetic
model*

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Dengfeng Liu(刘登峰)1,2,**, Hongtao Zhang(张洪涛)3, Zhizhong Lian(连治中)3, Baoguo Xu(徐保

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国)1

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1
Key Laboratory of Industrial Advanced Process Control for Light Industry of Ministry of Education,
Jiangnan University, Wuxi 214122, China
2
School of Internet of Things Engineering, Jiangnan University, Wuxi 214122, China

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3
Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi
214122, China
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Article history:
Received 16 October 2015
Received in revised form 17 February 2016
Accepted 26 March 2016
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* Supported by the National Natural Science Foundation of China (21276111, 21206053,

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61305017), 111 Project (B12018), Fundamental Research Funds for the Central Universities
(JUSRP11558), and the Fundamental Research Funds for the Central Universities (JUSRP51510).
** Corresponding author. liudf@jiangnan.edu.cn.
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Abstract Chinese rice wine making is a typical simultaneous saccharification and fermentation (SSF)

process. During the fermentation process, temperature is one of the key parameters which decide the

quality of Chinese rice wine. To optimize the SSF process for Chinese rice wine brewing, the effects
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of temperature on the kinetic parameters of yeast growth and ethanol production at various

temperatures were determined in batch cultures using a mathematical model. The kinetic parameters

as a function of temperature were evaluated using the software Origin8.0. Combing these functions

with the mathematical model, an appropriate form of the model equations for the SSF considering the

effects of temperature were developed. The kinetic parameters were found to fit the experimental data

satisfactorily with the developed temperature-dependent model. The temperature profile for

maximizing the ethanol production for rice wine fermentation was determined by genetic algorithm.

The optimum temperature profile began at a low temperature of 26°C up to 30 h. The operating

temperature increased rapidly to 31.9°C, and then decreased slowly to 18°C at 65 h. Thereafter, the

temperature was maintained at 18°C until the end of fermentation. A maximum ethanol production of

89.3 g/L was attained. Conceivably, our model would facilitate the improvement of Chinese rice wine

production at the industrial scale.

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Keywords Chinese rice wine, Temperature controlling, Simultaneous saccharification and

fermentation, Optimal temperature profile

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1 INTRODUCTION

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Chinese rice wine is a non-distillation beverage which is quite popular in south

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China [1]. So far, it is clear that temperature control in the Chinese rice wine brewing

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process is the most important controlling parameters governing the quality of Chinese

rice wine [2]. The temperature during Chinese rice wine fermentation affects the

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following sub-processes [3]: (1) Cell growth/death. During the first 20 h, temperature

affects cell growth and cell concentration affects ethanol production. (2) Starch
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hydrolysis. Temperature also affects the hydrolysis of starch from the sticky rice. (3)

Fermentation rate of ethanol production. Temperature affects cell growth, particularly

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for the Saccharomyces cerevisiae which transfers glucose to ethanol as we reported

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previously [2]. (4) Production of byproducts. Temperature affects the bacteria growth

which produces organic acids.

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However, the temperature control can be manually adjusted but it is necessary to

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determine the optimal temperature profile at various stages of the Chinese rice wine

fermentation process. To determine the optimal temperature profile, the development of

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the kinetic model for the fermentation process is needed [4, 5]. Thus far, although

several mechanistic models have been proposed for enzymatic hydrolysis of cellulose

[6, 7], ethanol fermentation by S. cerevisiae [8], the optimum temperature for

enzymatic hydrolysis of cellulose (typically 45–50°C) is higher than that of the

Chinese rice wine fermentation (30–35°C) with Saccharomyces cerevisiae as the

fermenting organism. However, during the ethanol production, the enzyme for their

study was purified cellulase and the temperature reached as high as 50°C. In addition,

most of their substrates were water-insoluble cellulose or starch flour. It is obviously

different from that for Chinese rice wine fermentation. During the Chinese rice wine

fermentation process, temperature cannot exceed 35°C, and the saccharification

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materials are whole steamed sticky rice. In addition, Chinese wheat qu instead of purify

enzymes were used in the process [4, 9, 10]. All of the above indicate that Chinese rice

wine fermentation is a new type SSF fermentation different from former ethanol

fermentation. Consequently, although there have several model-based control of

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temperature during simultaneous saccharification and fermentation (SSF) has been

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experimentally determined for industrial ethanol fermentation; new kinetic model

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suitable for Chinese rice wine fermentation and the optimal temperature profile is still

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needed to be developed.

Hitherto, there are three methods to construct kinetic model: (1) Mechanism

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Model; (2) Data-driven model and (3) Gray-box model. Recently, our group has

conducted a series of work to develop a suitable kinetic model for the Chinese rice
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wine fermentation based on the characteristics of Chinese rice wine brewing [11, 12].

Both ethanol and organic acids in the Chinese rice wine can affect its quality. Acquiring
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high ethanol concentration and keeping the organic acids at suitable concentration are
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the main objectives for the process [13]. Like any other process, the basic challenge in

Chinese rice wine brewing is to obtain high degree of hydrolysis of carbohydrate

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substrates together with high ethanol yield.

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In this work, kinetic parameters were estimated from the kinetic model developed

by our group, the temperature-dependent model has been developed and validated
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using the fermentation data with the industrial strain of the yeast S. cerevisiae Su-25 at

various temperatures with pretreated whole stick rice. Subsequently, an optimal

temperature profile for the SSF process of Chinese rice wine brewing to acquire high

content of ethanol was determined based on the fitted model.

2 MATERIALS AND METHODS

2.1 Mathematical model

The model includes temporal variations of the concentrations of starch (S), maltose

(M), maltotriose (R), yeast cells (C), ethanol (A), dissolved oxygen (O), wheat qu (E), and

starch-qu complex (C1).

The mathematical model developed in this work was constructed based on our

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previous report [12]. As we describe in the kinetic model sections, the concentration of

glucose were always low during the simultaneous saccharification and fermentation (SSF)

process. At first, we hypothesized that the glucose was rapidly utilized by the yeast which

was why the glucose level was always low. To verify our hypothesis, saccharification

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experiment was conducted. The result showed that the concentration of glucose was also as

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low as 3.5 g/L in saccharification experiment only using wheat qu and starch, which was

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similar to that of under SSF condition. Therefore, during the Chinese rice wine

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fermentation process, maltose and maltotriose were the main sugar used by Saccharomyces

cerevisiae Su-25 to produce ethanol and other by-products.

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Our mathematical model is shown as follows based our previous publication [12]:

dS
 k1SE (1)
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dt

dR R O
 1.037k2C1  k4 C (2)
dt K s1  R K s 2  O
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dM M O M
 1.056k3C1  k5 C  k6 C (3)
dt K s3  M K s 2  O Ks4  M
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dC R O M O
 0.084k 4 C  0.088k 5 C (4)
dt K s1  R K s 2  O K s3  M K s 2  O
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dA M
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 0.54k 6 C (5)
dt K s4  M

dO R O M O
 k 4 C  k5 C  Oin (6)
dt K s1  R K s 2  O K s3  M K s 2  O
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 k1SE  k 2  k3 C1
dE
(7)
dt

 k1SE  k 2  k 3 C1
dC1
(8)
dt
where the constants (1.037, 1.056, and 0.54) are the yield coefficients accounting for the

molecular weight differences among the products; the other constants are obtained from the

Balling equations [14]; Ks1-Ks4 are the saturation constants of each substrate.

2.2 Materials

The Chinese wheat qu used in this study was supplied by the Shaoxing Nverhong Rice

Wine Company (Zhejiang, China) and stored at room temperature. Sticky (glutinous) rice

from northern China, was purchased from the Vanguard Market, Wuxi, China. Yeast strains

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Saccharomyces cerevisiae Su-25 supplied by the Nverhong Wine Company (Shaoxing,

Zhejiang, China) was used in this study and stored at 4°C.

2.3 Thermo-controlled fermentation process

Rice wine fermentation experiments were carried out with rice wine yeast strains

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Saccharomyces cerevisiae Su-25 and wheat qu.

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The yeast strain was stored at 4°C on slants of YPD agar medium. The yeast inoculum

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was transferred to a new slant of YPD agar and cultured for 24 h at 28°C. A sloop of yeast

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culture was added to 50 mL YPD broth (250 mL flask) and cultured at 28°C for 18 h. Based

on the 1:10 ratio, yeast seed inoculum to new YPD broth (100 mL in 500 mL flask) and

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cultured at 28°C for another 18 h.

The experiments were conducted in 7-L tank fermenters (BioFlow/CelliGen 115, New
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Brunswick Scientific, Enfield, CT, USA) containing about 5 L batch fermentation broth

(1200 g steamed rice (dry weight), 204 g wheat qu, 120 ml yeast seed culture and 2400 ml

tap water) at constant temperature for 96 h, four temperatures (18°C, 23°C, 28°C, 33°C)
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were chosen.
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2.4 Analytical methods

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At the predetermined time, samples of 2 mL cultures were taken from the fermentor
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for analyses with HPLC using the method describing by Liu et al. [3, 11] as follows: 2 mL

of samples from each experiment were taken out, and then centrifuged and filtered through
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0.45-μm-pore Durapore (PVDF) membrane filters (Fisher, Pittsburgh, PA, USA),

subsequently analyzed with high-performance liquid chromatography (HPLC). Agilent

HPLC system (Agilent 1200 Series, Santa Clara, CA, USA) were used for injection and

detection. An Aminex HPX-87H column (300×7.8 mm) (Bio-Rad Labs, Richmond, CA,

USA) was used to identify and determine the levels of sugars (maltotriose, maltose and

glucose) and ethanol with G1314B VWD detector and a G1362A RID detector (Agilent

1200 Series, Santa Clara, CA, USA). The HPLC conditions used were as follows: flow rate

was 0.5 mL/min, the mobile phase consisted of 60-mL acetonitrile and 1270-μL sulfuric

acid for every 1000-mL solvent, injection volume was 20 μL per fixed loop.

2.5 Determination of model parameters and its validation

To fit the data to the model, the parameters in the model were estimated using a

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nonlinear least square method which minimizes an objective function of Z [15]:

n
Z X   ( X i ,exp  X i , sim ) 2
i 1 (9)
where n represents the number of data points, ZX, represent the least squares discrepancies

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between experimental data and simulated results of biomass, X represent starch, maltose,

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yeast cells, ethanol, dissolved oxygen, wheat qu, and starch-enzyme complex, respectively.

And i denotes the number of experimental data points. ‘exp’ and ‘sim’ denote the observed

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value, and simulated value, respectively. The model was constructed by using fourth-order

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Runge-Kutta method in MATLAB. MSE (mean squares error) and R2 were used to evaluate

the experimental data and modeling data. The MSE value can be calculated by the sum

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squares errors divided by the length of actual data period [16].

1  yi  f i 
iNT
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2
MSE  (10)
nt

  yi  fi 
2
R2  1 
 2 (11)
 yi  y
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where f i , y i , y and nt are the model data, experimental data, mean experimental
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data and length of actual data period, respectively.

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2.6 Development of temperature-dependent model and its validation

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Kinetic parameters as a function of temperature in the range from 18 to 33°C were

estimated using the software package of Origin 8.0. These functions coupling with the
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fermentation kinetic model of Chinese rice wine fermentation were described in 2.1. A

mathematical model was developed for the Chinese rice wine fermentation. This model

considers the effects of temperature and it was verified with existing data.

2.7 Optimization of temperature profile

The optimal temperature profile for maximizing ethanol production was determined

using genetic algorithms (GAs) with the developed model in 2.6 sections. We pick the

'PopulationSize', 'Generations', 'StallTimeLimit', 'StallGenLimit', and 'TolFun', as 200, 100,

Inf, 2, and 1e-6 to run the GAs. The objective function was chosen as Equation (11) for

maximizing ethanol production by optimizing the temperature profile.

min J   A(t f ) (12)

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Where A(tf) is the ethanol concentration at the end of fermentation.

3 RESULTS AND DISCUSSION

3.1 Selection of key controlling variables and refinement of model

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To construct model of the fermentation process and setup automatic control parameter,

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the most important procedure is determining the key controlling variables. For the Chinese

rice wine production based on the China national standard GB-13662-2000 which can be

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classified into four types: dry-type (sugar content ≤15 g/L), half dry-type (15 g/L<sugar

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content ≤40 g/L), half-sweet type (40 g/L<sugar content≤100 g/L) and sweet-type (sugar

content>100 g/L).

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To evaluate the effect of temperature on ethanol and sugar production, Chinese rice

wine fermentation was carried out at 18°C, 23°C, 28°C, and 33°C. The results are shown in
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Fig. 1. The concentration of maltotriose at 33°C increased gradually until fermentation end,

and the concentration of maltotriose at 18°C arrived at the highest concentration of 10.3 g/L
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and then gradually reduced to 8.4 g/L at 140 h. For the concentration of maltotriose at 28
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and 23°C, both of them all arrived at the highest concentration at 45 g/L and following

gradually reduced until 140 h (Fig. 1A). The concentration of maltose reached the highest
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capacity of 90 g/L at 25 h and it remained unchanged until 40 h. Then, it began to reduce

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gradually. The concentration of maltose increased at the highest concentration of 79.5 g/L at

25 h and then reduced quickly, and all arrived at lower values at 80 h. However, the
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concentration of maltose arrived at 80 g/L at 20 h, it reduced to 40 g/L at 25 h and then

increased to 70 g/L, following reduced gradually until fermentation end (Fig. 1B). But the

concentration of glucose only went up to around 3 g/L (Fig. 1C). Consequently, maltose

and maltotriose were chosen as variables but glucose was not considered in the resulting

model.

For ethanol, from Fig.1D, the final concentration of ethanol in Chinese rice wine broth

increased with temperature increasing from 18 to 28°C, and dropped to 30 g/L at 33°C.

These results reveal that temperature control is an important parameter for ethanol

production for Chinese rice wine brewing. Similar results were also reported for industrial

ethanol fermentations [17, 18]. Consequently, ethanol, maltose and maltotriose were

considered as the major controlling parameters.

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A) B)
18 (b) 90
16 (a) 16
100 80 80
14
40 70

Maltotriose (g/L)
12
3 12

(g/L) (g/L)
60 40 60
3 10 75 100 100 50

GlucoseMaltose
8 8 4030 40 40

Ethanol (g/L)
3 6 30

Yeast (g/L)
2 75 30

Glucose (g/L)
50 75

Ethanol (g/L)
4 4

Yeast (g/L)
2 20 20
20 30

Glucose (g/L)
2

Ethanol (g/L)
50 10

Yeast (g/L)
2 25 50 0 20
10 0 0
0 20 40 60 80 100 120 140
0 20 40 60 80 100 120 140

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1 10 20 25
Time (h) Time (h)
0 25 10
1
C) 0 D) 0
0 20 400 60 80 100 0 10

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5 Time (h)
0 20 405.0 60 0 80 100 0
4.5 (h) 12 o 12
Yeast
0 Glucose Time
Ethanol Model 18oC
4 0 20 40 60 80 4.0100
Glucose 10 0 23oC 10
Yeast Ethanol Model
Time (h) 3.5 28 C

Ethanol (%, v/v)

Glucose (g/L)

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3 Yeast Glucose Ethanol
3.0 8Model33 C 8

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(c) 2.5 6 6
2 2.0
1.5 4 4

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1 1.0 2 2
0.5
0 0.0 0 0
0 20 40 60 80 100 120 140 0 20 40 60 80 100 120 140
Time (h) Time (h)

Figure 1 The time course of the concentrations (g/L) of maltotriose, maltose, glucose, and

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ethanol at (A) 18°C, (B) 23°C, (C) 28°C, (D) 33°C of Chinese rice wine fermentation production

3.2 Development of the temperature-dependent model

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To develop the model and validate the temperature profile for the SSF process, approaches
described by Fig. 2 were used. The experimental data were utilized for parameter
estimation with our accordingly developed kinetic model in this report. The estimated
parameters from experimental data with our kinetic model are provided in Table 1. The
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parameter sensitivity test was conducted to examine the influence of the model parameters.
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Changes of ±10% were introduced to each parameter from its optimized value for the first
ambient-temperature run and the ratio of the total sum of squared changes in the outputs
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(products) over the parameter change was computed as an indicator of the influence of the
parameter or parameter sensitivity. The result revealed that k1, k2, k3, k5 and k6 are sensitive.
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Table 1 Estimated parameter values at various temperatures during Chinese wine fermentation
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Temperature k1 k2 k3 k4 k5 k6

18°C 1.362 16.814 0.367 0.081 1.418 0.412

23°C 1.623 19.812 0.591 0.562 2.631 0.812

28°C 1.695 20.818 0.685 0.643 2.929 0.886
33°C 2.594 20.810 1.334 0.164 2.946 1.462

The temperature is an important parameter for ethanol fermentation. The reaction rate

of enzyme for starch hydrolysis to produce maltose and maltotriose are also affected

accordingly.

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Mathematical model development Parameter estimation for rice wine

for rice wine fermentation process fermentation process

Experiments for rice wine fermentation

under different temperature separately

Parameters described as
a function of temperature

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Temperature dependent mathematial

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model for rice wine fermentation process

Optimization of temperature profile for

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maximum rice wine production

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Figure 2 Schematics for modeling and validating the temperature profile of the SSF process for
Chinese rice wine

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The effect of temperature on the specific hydrolysis rate of starch [19] and ethanol

production [20] are quantitatively described by the Arrhenius equation which has been used
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to describe ethanol production with cellulose: ki(T1) = ki(T2)exp[–ΔH/R(1/T1–1/T2)] in

which ki(T1) and ki(T2) are the rate constant at the absolute temperature T 1 and T2; R is the

gas constant (8.314×10-3 kJ/[mol·K]); and ΔH is the activation enthalpy of the reaction
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described by rate coefficient ki (kJ/[mol·K]). We obtained ΔH of 33.018 (kJ/[mol·K]) and

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55.82684(kJ/[mol·K]) for k1 and k6, estimated using Origin 8.0, respectively. Parameters k2,
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k3, k4 and k5 are used in model to describe maltotriose changing. Using the parameter values
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given in Table 1, kinetic parameters as a function of temperature were developed using

Origin 8.0, the resulting model is shown in Table 2. Good agreement has been found
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between the developed function and the kinetic parameters (shown as Fig. 3).

Fig. 3 shows that k1 and k6 have the similar tendency within the temperature range

18-33°C. The possible reason is that these parameters are used to describe fermentation

kinetic with the Arrhenius equation and the reaction mechanism is the same. In addition, k1

and k6, as well as, k2 and k5 have the same tendency. k4 gradually increased and then

decreased. And k3 increased with the temperature increasing (Fig.3).

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A) B)
A)
A) 3.0
B) B) 25
(b)
(a) 18 (b)
2.8 100
16 (a) 16
2.6 40 100
32.4 14
40 40

Maltotriose (g/L)
12
3 75 100 20 12
2.2 3100 40
75 100 30 40 3 10

k1 (g/L/h)

(g/L/h)
100

(g/L)
2.0 3

k(g/L)
8 30 40 8 30

(g/L)
21.8 75

(g/L)

(g/L)
50 75

(g/L)
3

(g/L)
6 30

2
(g/L)
2 50 75 20 30 15 75

(g/L)
2

(g/L)
Glucose
1.6

(g/L)
Ethanol
(g/L)
4 4

(g/L)
Yeast
2

Glucose

(g/L)
50

Glucose
1.4 2

Ethanol
20 30

Ethanol
20

(g/L)
25 50

(g/L)
Yeast
2

Yeast

(g/L)
11.2 50

Glucose

Ethanol
(g/L)
2 25 50 20

Glucose
Yeast
10 1 0

Ethanol
1.00 20 40 60 80 100 120 140 10 20 10 25

Yeast

Glucose
15 20 1 25 30 35
0 25 10 20 15 25 20 25 30 35

Ethanol
10

Yeast
Time (h) o
0 1

T
Temperature ( C) 0 D)25 0 10 Temperature ( C)
o

1
C) 0 0 20 400 60 80 100 D) 0 10 0
C) 0 20 Time (h)
40 0D) 10100
C) 0.010Yeast 0 060 80
20 100 60 0 80
40
D) 0.8
0
0 Time (h)
Glucose 0 20Ethanol
40 60 0 Model
Time (h) 80 100 0

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0 0
Yeast 20Glucose
40
Yeast 60 Ethanol 80 Time100
Glucose (h) Model 0
Ethanol Model
0 20 40
Time 60 80 100
0.008 Yeast(h) Glucose 0
Ethanol0.6 Model
Yeast Time (h)
Glucose Ethanol Model
(c) Yeast Glucose Ethanol Model
0.006(c)
k3 (g/L/h)

k4(g/L/h)
0.4

R
0.004
0.2

0.002

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0.0
15 20 25 30 35 15 20 25 30 35
o o
Temperature ( C) Temperature ( C)

E) F)
E) F) 1.6
3.4
3.2 1.4
3.0
1.2
2.8

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2.6 1.0
2.4

k6(g/L/h)
k5(g/L/h)

2.2 0.8
2.0 0.6
1.8
1.6 0.4
1.4
0.2
1.2
1.0 0.0
15 20 25 30 35 15 20 25 30 35
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o o
Temperature ( C) Temperature ( C)

Figure 3 Effect of temperature on the kinetic parameters (A) k1; (B) k2; (C) k3; (D) k4; (E) k5; (F)
k6 during Chinese wine fermentation
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Table 2 Evaluation of the Kinetic Parameters as a Function of Temperature (T) during Chinese
wine fermentation
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Parameters Function
k1   33.018 1 1 
1.0608  10 6 exp  (  )
 8.314  10
3
273  T 306 
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k2 -5.67058+1.79294×T-0.03006×T2
k3 - 0.00166 + 3.2 ×10-4 × T
-5.7482+0.4962×T-0.0096×T2
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k4
k5 -7.41206+0.7076×T-0.01196×T2
k6   55.82684 1 1 
4.85324 10 9 exp  (  )
 8.314 10
3
273  T 306 
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3.3 Validation of the resulting temperature model with experimental

data

After we developed the kinetic parameters as a function of temperature, the

temperature-dependent model for describing the Chinese rice wine fermentation was

worked out. However, to optimize the Chinese rice wine fermentation, we need to validate

the model with the experimental data and the parameters summarized in Table 1.

Using the parameter function in Table 2, the data from the developed

temperature-dependent model were shown as Fig. 4, in which indicates that a good

agreement is found between the model and the experimental data for maltose and

maltotriose concentrations, as well as ethanol production. In the Chinese rice wine

fermentation broth, analysis of the cell concentration is still difficult. Therefore, in Fig. 4,

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the experimental data does not show the cell concentration. In addition, as Chinese rice

wine is a semi-solid fermentation, the dissolved oxygen (DO) is difficult to determine

accurately. The comparison of the ethanol concentrations of the experimental data and

model at various temperatures are shown in Table 3. We concluded that all the parameters

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of the developed model in this report are applicable to the SSF process besides the values at

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23°C. It is clear that the experimental data on ethanol level are higher than that of model

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predicted. The reason is that yeast growth is more robust at 23°C，and has longer viability

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time at 23°C than other temperatures. Consequently, higher ethanol level is produced by the

yeast at 23°C.

A)
80
16 (a) Maltotriose
18
16
180
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B)
100
90
(b) 90
80
180
(g/L)Ethanol, Yeast (g/L)

Maltotriose, Ethanol, Yeast (g/L)

Ethanol
70 160
100 160
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Sim-Yeast
Sim-Maltotriose 14
140 40 80 70 140
Maltotriose (g/L)

Sim-Ethanol
3 60
12 Maltose 12
(g/L) (g/L)
Maltose, Starch (g/L)

Maltose, Starch (g/L)

Sim-Maltose 60 70 40 60
Sim-Starch 120 120
50 3 10 75 100 60 100 50
100 100
GlucoseMaltose

8
40
8 4030 40 50 40
(g/L)

3 6 80 30 80
2 75 30
Glucose (g/L)

50 75
Maltotriose,

40
Ethanol (g/L)
30
4 4
Yeast (g/L)

2 60 20 20 60
30
Ethanol

20 30
Glucose (g/L)

20
Yeast

2
Ethanol (g/L)

40 20 50 10 40
Yeast (g/L)

2 25 50 0 20
1010 20 0 0 20
0 20 40 60 80 100 120 140 0 2010 40 60 80 100 120 140
0 1 0 10 20 0 25 0
D

0 10 20 30 Time
40 (h)
50 60 70 80 90 100 0 10 Time
20 (h)
30 40 50 60 70 80 90 100
0 25 10
1 Time (h) Time (h)
0 0
0 20 400 60 80 100 0 10
C) 5 Time (h) 0 20 405.0 60 0 80D) 100 0
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100 180 12 12
Model 1835oC 180
o
Yeast
0 Glucose Ethanol 4.5 (h)
Time
4 90 0 20 40 60 80 4.0100160
Maltotriose, Ethanol, Yeast (g/L)

Maltotriose, Ethanol, Yeast (g/L)

Yeast Glucose 10 0 2330oC

Ethanol Model 10 160
80 Time (h) 3.5 140 28 C
v/v)

140
Glucose (g/L)

o
Glucose 8Model33
25C 8
(g/L)

Maltose, Starch (g/L)

3 70 Yeast Ethanol
3.0
120 120
(c)
(%,

60 2.5 6 6
Starch

100 20 100
Ethanol

2 50 2.0
1.5 80 4 15 4 80
Maltose,

40
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1 30 1.0 60
2 2 60
10
0.5 40 40
20
0 0.0 0 5 0
10
0 20 40 60 80 100 120 140 20 0 20 40 60 80 100 120 140 20
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0 Time (h) 0 0 Time (h) 0

0 10 20 30 40 50 60 70 80 90 100 0 10 20 30 40 50 60 70 80 90 100

Time (h) Time (h)

Figure 4 Experimental data and model simulations for ethanol production with the
development temperature-dependent kinetic model at various temperatures
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Table 3 The comparison of experimental and model predicted ethanol concentration for various
temperatures
Temperature model ethanol concentration (g/L) at the end of 100 h
(°C) Model predicted Experiment value
18 69.23 66.46
23 78.15 97.72
28 78.18 79.78
33 27.01 26.63

3.4 Optimization of the temperature for maximizing ethanol production

Rice wine fermentation is a typical nonlinear system. Genetic algorithm is effective

for complex nonlinear system optimization. Therefore, an optimization control strategy of

GAs was developed to acquire optimal temperature profiles.

Optimization of the temperature for non-isothermal SSF process was made using the

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optimal profile. The prediction of the stated variables such as maltotriose, maltose and

ethanol are shown as different type lines in Fig. 5A. The model predicted the temperature

profile was nonlinear (Fig. 5B).

A)

T
100 (b)100
90 3
(a) 10040 40 180

Maltotriose, Ethanol, Yeast (g/L)

3Opti Yeast C)100 75 40

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Opti Maltotriose
3
80 Opti Ethanol 160

Maltose, Starch (g/L)

75
Opti Maltose 75 30 30

Glucose (g/L)

(g/L)
30

Ethanol (g/L)

Ethanol (g/L)
Glucose (g/L)
2Opti Starch2

Ethanol (g/L)
140

Yeast (g/L)
70 2

Yeast (g/L)

Yeast (g/L)
5050
(c) 50

Glucose
20 20
60 25
20 120
1
50 1
1 25 25 10 D) 100

R
0 10
0 10
40 0 20 40 60 80 0 100
0 0 80
0 Time (h)
30 0 0 20
60 Glucose 40
80 100Ethanol 0 Model
Yeast
0 20 40 Time60 (h) 80 100 0 60
20 Yeast TimeGlucose
(h) Ethanol Model

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Yeast Glucose Ethanol Model 40
10
20
0
35 0
Optimized curve
Fit curve
B)
30

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Temperature ( oC )

25

20
MA
15
0 20 40 60 80 100
Time (h)

Figure 5 Optimized temperature profile and ethanol production by SSF during Chinese wine
fermentation
D

The temperature profile estimated by Origin 8.0 can be described by the equation
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shown below:

T  26.66642  0.08933 t  0.038  t 2  0.00144  t 3  1.76311105  t 4  7.05567 108  t 5 (13)

P

Where, t is time. Temperature profiling begins with the low temperature at 26°C, which
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allows the growth of the yeast cells during the first 20 h. Subsequently, the operating

temperature increased rapidly up to 31.97°C at 30 h, after which it decreased slowly to

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18°C at 65 h. The predicted final concentration of ethanol was 89.3 g/L, which is 14–21%

higher than that of the traditional SSF process (manual controlling).

4. CONCLUSION

The effect of constant temperature on Chinese rice wine fermentation and optimal

temperature profile was investigated at a scale-down level using Mathematical model was

developed ematical model. The operating temperature of Chinese rice wine fermentation

was restricted by the temperature for the reason that high temperature will cause excessive

organic acid production. Thus, the upper and lower limits of operating temperature are 18

and 33°C, respectively, in this work. The accordingly developed temperature-dependent

model has good agreement with the experimental data. Our model suggests that temperature

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has a strong effect on starch hydrolysis as well as cell growth and ethanol production. The

optimal temperature profile is: during the first 20 h the temperature increases gradually

from 26°C to 31.9°C. The operating temperature at around 31.5°C lasted for 13 h.

Thereafter, it decreases slowly to 18°C at 68 h. The predicted final concentration of ethanol

T
is 89.3 g/L. Our model conceivably can facilitate the improvement of the production of

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Chinese rice wine at the industrial scale.

R
NOMENCLATURE

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S Starch concentration, g·L-1

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R Maltotriose concentration, g·L-1

M Maltose concentration, g·L-1

MA
𝐸 Qu concentration, g·L-1
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C1 Starch- qu complex concentration, g·L-1

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C Cell mass concentration, g·L-1

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A Ethanol concentration, g·L-1

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O Oxygen concentration, g·L-1

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k1-k6 Chemical reaction rates, g·L-1·h-1

Ks1-Ks6 Saturation constant, g·L-1

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