762 Biotechnol. Prog.

1997, 13, 762−767

Kinetics of Simultaneous Saccharification and Lactic Acid

Fermentation Processes
Jun Luo, Liming Xia, Jianping Lin, and Peilin Cen*
Department of Chemical Engineering, Zhejiang University, Hangzhou 310027, People’s Republic of China

After pretreatment of corn cob by dilute acid, the lignocellulosic residue was used as
raw materials for the simultaneous saccharification and lactic acid fermentation
(SSLF). Because of the same optimal temperature and pH requirement as well as
the anaerobic condition, the lactic acid fermentation is perfectly compatible with
enzymatic hydrolysis of cellulosic materials. In the SSLF processes, the final
concentration of lactic acid reached 33.97g/L with a conversion ratio of 79% based on
the consumed cellulose. A mathematical model is suggested to simulate the SSLF
process with good agreement.

Introduction substrate for microorganism growth, maintenance, and

It is a popular process in China to produce xylose from
corn cob by dilute acid hydrolyzation methods (You,
1984). The residue is mainly lignocellulosic material.
How to treat the residue is an important issue either from
comprehensive utilization of the renewable resources or
from environmental protection points of view. The
advantage in using the corn-cob residue is that the
residue can be enzymatically hydrolyzed without further
pretreatment.
The simultaneous saccharification and fermentation
(SSF) process is an effective process in which enzymatic
hydrolysis of lignocellulosic materials and fermentation
are coupled together to eliminate the inhibition caused
by cellubiose and glucose (Enari, 1983) and to combine
the two steps into one. There is a lot of research on the
SSF processes focused on the production of ethanol
(Spindler and Wyman, 1989) and single cell protein, SCP
(Xu et al., 1995). The optimal temperature for enzymatic
hydrolysis is 50 °C, but 30-35 °C for ethanol and SCP
fermentation, which means the two processes are not
compatible in operation conditions. The yield of ethanol
or SCP from glucose is only about 50%. Therefore, a new
SSF process should be exploited.
Lactic acid is an important organic acid that is widely
applied in food and chemical industries and a prospective
candidate for making the biodegradable polymers (Lip-
insky and Sinclair, 1986). The lactic acid fermentation
is carried out at 50 °C, at pH 5.0, and at anaerobic
condition (Jing et al.; 1995), which is the same as that
for enzymatic hydrolyzation of cellulose, and the theoreti-
cal yield of lactic acid is 100% on the basis of the glucose
consumed. It seems that the simultaneous saccarifica-
tion and lactic acid fermentation (SSLF) process should
be more compatible and comprehensive for the utilization
of renewable resources. The SSF processes for lactic acid
production from pure cellulose had being reported with
a yield of 60% (Takagi, 1984; Abe and Takagi, 1991).
In the SSF process, both enzymatic hydrolysis and
fermentation must be evaluated simultaneously in the
mathematical model. For enzymatic hydrolysis of cel-
lulosic materials, a kinetic model must include multien-
zyme catalysis with product inhibition in a multiphase
system (Lee and Fan, 1980; Dekker, 1986; Beltrame et
al., 1984). The hydrolytic product, glucose, is used as the
* To whom all correspondence should be addressed.
S8756-7938(97)00100-8 CCC: $14.00 © 1997 American Chemical Society and American Institute of Chemical Engineers
product formation simultaneously. A heterogeneous
multienzyme catalytic model with ethanol inhibition was
suggested to describe the SSF process for ethanol produc-
tion (Philippidis et al., 1993), but cell growth and ethanol
formation were not included in the model. On the basis
of the assumption that the rate of hydrolysis per adsorbed
enzyme is a strong function of substrate conversion,
competitive inhibition of cellubiose hydrolysis by β-glu-
cosidase, and ethanol inhibition of Saccharomyces cer-
evisiae growth, a mathematical model was proposed to
analyze the conversion of particulate biomass to ethanol
in continuous solid retaining and cascade bioreactor
(South and Lynd, 1994). The kinetic models for lactic
acid fermentation have been widely studied in the
literature (Gonclaves et al., 1991); however, the math-
ematical model for the SSLF process has not been
reported in the literature.
The purposes of this work are to exam the compatibility
between enzymatic saccharification of corn-cob residue
and lactic acid fermentation, to optimize operation condi-
tions for high lactic acid yield, and to establish a
comprehensive kinetic model for the SSLF process.
Materials and Methods
Microorganisms. Cellulases were produced in this
laboratory by using either Trichocherium reesei (origi-
nally from Rut C30) or Aspergillus niger(originally from
Zhejiang Microbiology Research Institute). The crude
enzyme solution was applied in the SSLF process. Lactic
acid was produced by a strain of Lactobacillus delbrium
(from Zhejiang Microbiology Research Institute).
Corn-Cob Residue from Xylose Production. Corn-
cob residue obtained from a xylose production factory or
made in the laboratory was used as the raw material. In
the laboratory corn cob was ground into particles less
than 3 mm in diameter and then was hydrolyzed at 106
°C for 4 h with 1% H2SO4 (liquid/solid ratio is about 8:1).
The supernatant can be used for xylose production, and
the corn-cob residue is the substrate for the SSLF
process. The composition of the corn-cob residue was as
follows: cellulose, 62%, hemicellulose, 19%; lignin, 13%.
Medium. Medium for T. reesei. The solid medium
was potato dextrose agar. The liquid culture medium
was as follows (g/L): corn cob residue, 70.0; glucose, 1.0;
peptone, 1.0; wheat bran, 3.0; salt solution, 10.0 mL. The
salt solution consists of the following (g/L): (NH4)2SO4,
Biotechnol. Prog., 1997, Vol. 13, No. 6
Table 1. Activities of Cellulases in the Final
Fermentation Broth
name of strain Et, (FPIU/mL) EB, (BU/mL)
T. reesei 3.2-4.1 0.55-0.65
A. niger 1.1-1.4 5.20-6.25
14.0; urea, 3.0; MgSO4‚7H2O, 3.0; KH2PO4, 20.0; CaCl2‚
2H2O, 4.0; FeSO4‚7H2O, 0.05; MnSO4‚H2O, 0.016; ZnSO4‚
7H2O, 0.014; CoCl2‚6H2O, 0.037. The pH was controlled
by adding 50 mL of 1 M citric acid buffer solution.
Medium for A. niger. The solid medium was potato
dextrose agar. The liquid culture medium was as follows
(g/L): corn-cob residue, 30.0; wheat bran, 10.0; (NH4)2-
SO4, 1.0; urea, 0.35; MgSO4‚7H2O, 0.35; KH2PO4, 2.5;
CoCl2‚6H2O, 0.35.
Medium for L. delbrium. The medium for seed
preservation was 20% defatted milk powder.
The liquid medium for lactic acid fermentation was as
follows (g/L): yeast extract, 1.0; peptone, 1.0; NaCl, 0.1;
and glucose as required. The pH was regulated to pH
4.8-5.0 by adding CaCO3.
The liquid medium for the SSLF process was as follows
(g/L): yeast extract, 1.0; peptone, 1.0; KH2PO4, 0.1; Na2-
HPO4, 0.1; MgSO4‚7H2O, 0.05; salt solution, 2.0 mL. The
salt solution consists of the following (g/L): FeSO4‚7H2O,
0.105; CuSO4‚5H2O, 0.17; MnSO4‚H2O, 0.0144; ZnSO4‚
7H2O, 0.882; CoCl2‚6H2O, 0.05. Corn-cob residue was
added as the experiment required, and CaCO3 was used
to regulate the pH value of the fermentation broth to a
range of pH 4.8-5.0.
Experiments. Both cellulase production and the
SSLF process for lactic acid production were carried out
in a Biostat-B 2-L stirred-tank fermentor (B. Braun). In
cellulase production, the operation conditions were con-
trolled at 28 °C, 300 rpm, and an aeration rate of 0.3
vvm. The inoculum ratio was 10%. The initial pH value
in the broth was regulated to pH 4.5-5.0. The pH value
in the fermentation broth would gradually rise to about
pH 7.0-7.5, which indicated the end of the fermentation
process. During the SSLF process, the temperature was
kept at 50 °C and pH was regulated in the range of pH
4.8-5.0 by adding CaCO3. The cellulase solution and L.
delbrium were inoculated simultaneously. The rotating
rate of the stirrer was 200 rpm to offer sufficient mixing
without aeration.
Analysis Methods. Glucose and cellubiose are ana-
lyzed by the glucose oxidase-peroxidase method (Berg-
meyer, 1974) and by the HPLC method, respectively;
lactic acid is analyzed by the Barker-Summerson method
(Barker and Summerson, 1941). The total cellulase
activity is measured by filter-paper assay and β-glycosi-
dase activity by a β-glycosidase assay (Ghose, 1987).
Results and Discussions
Preparation of Cellulases. The preparation of cel-
lulases from either T. reesei or A. niger was carried out
in a Biostat-B 2-L fermentor. The batch fermentation
period was about 36-48 h. The ranges of final activities
of cellulase (Et) and β-glucosidase (EB) at different batch
fermentation are listed in Table 1. The whole fermenta-
tion broth containing crude cellulases was applied in the
SSLF processes without any purification.
Hydrolysis of Corn-Cob Residue by Cellulases.
The typical results of the hydrolysis of corn-cob residue
by cellulases are shown in Figure 1. It is obvious that,
with the accumulation of both cellubiose and glucose, the
rate of hydrolysis is inhibited and decreases fast; only
about two-thirds of the cellulose is hydrolyzed.
763
Figure 1. Time course of enzymatic hydrolysis of corn-cob
residue (initial activity of cellulases is 10.0 FPIU/gS): 0,
cellulose; 2, glucose; O, cellubiose; s, calculated curve.
Table 2. The Effects of Cellulase Activity on the SSLF
Process (Initial Cellulose Concentration ) 43.4 g/L)a
for given EC0 (FPIU/gS)
1.8 2.0 2.4 3.0 4.0 5.0 8.0 10.0
P (g/L) 7.03 10.95 10.81 14.18 23.61 31.56 34.20 36.70
Y (%) 16 25 25 32 54 72 80 84
a The cellulase is from T. reesei.
Table 3. Effects of Substrate Concentration on the SSLF
Process (EC0 ) 5 IU/gS, from T. reesei)
for given Ca (g/L)
20 60 100 150 200
P (g/L) 8.6 20.8 31.7 35.0 42.2
Y (%) 98 79 72 53 48
a C is the concentration of the corn-cob residue, g/L.
Effects of the Initial Activity of the Cellulases
and the Concentration of the Substrate on the
SSLF Process. The effects of the initial cellulases
activity and the substrate concentration on the SSLF
process were evaluated, and the results are listed in
Tables 2 and 3, respectively. It is obvious that, with the
increase of cellulase activity, the lactic acid yield in-
creases too. But, with the increase of substrate concen-
tration, the yield of lactic acid drops fast.
The increase of the cellulase activity will raise the cost
of the enzymes, and the low substrate concentration will
reduce the productivity and increase the cost for separa-
tion. Both factors are unfavorable. In order to optimize
the process, a relative factor F was defined as follows:
F ) EC2(1 - C0)/C0 (1)
The relationship between F and the lactic acid yield Y
is shown in Figure 2. If the costs of enzyme, substrate
and separation process as well as the sell price of the
lactic acid are known, then we can use eq 1 and Figure
2 to determine the best values of Y, EC, and C0. For
example, if Y ) 70%, the F value should be about 500
from Figure 2. Then if C0 ) 0.05, the value of EC should
be about 5.1 IU/gS. Also from Figure 2, it is obvious that
there is a wide range of operation conditions in which
the lactic acid yield is about 80%.
Effects of the Activity of β-Glycosidase. The
cellulase is a mixture of at least three enzymes: endo-
β-1,4-glucanase, exo-β-1,4-cellobiohydrolase, and β-gly-
cosidase. The β-glycosidase activity in the cellulases
produced by T. reesei is lower than that by A. niger. In
cellulose hydrolyzation process, the addition of β-glycosi-
dase can reduce the concentration of cellubiose and favor
the production of glucose. Table 4 shows the experimen-
764 Biotechnol. Prog., 1997, Vol. 13, No. 6

Figure 2. Relationship between F and the lactic acid yield.

Figure 4. Comparisons of experimenal data and calculated
Table 4. Effects of the Ratio of Cellulases from T. reesei curves of the SSLF process, C0 ) 42.7, EB0 ) 0.1, E0 ) 0.3, and
to That from A. niger (EC0 ) 5 FPIU/gS, C0 ) 43.4 g/L) X0 ) 0.12; cellulose, 0; lactic acid, 2; glucose, 4; cellubiose, O;
for given ratio (V/V) biomass, b.

10:0 9:1 8:2 6:4

P (g/L) 32.32 35.06 30.15 20.28
Y (%) 74 80 70 47

Figure 5. Simplified mechanism of the SSLF process.

Figure 3. Comparisons between experimenal data and calcu-

lated curves of the SSLF process, C0 ) 42.7, EB0 ) 0.05, E0 )
0.3, and X0 ) 0.14: cellulose, 0; lactic acid, 2; glucose, 4;
cellubiose, O; biomass, b.

tal results with the mixed cellulases produced by T. reesei

and A. niger, respectively, at different ratios. When the
ratio is 9:1 (cellulases from T. reesei to that from A. niger), Figure 6. Adsorption isotherm of cellulases on the corn-cob
the yield of lactic acid is the highest. A further decrease residue at 50 °C: 0, experimental data; s, calculated curve.
of the ratio will cause a sharp drop of the lactic acid yield
because of insufficient activities of endo-β-1,4-glucanase Adsorption of Enzyme EC on Corn-Cob Residue.
and exo-β-1,4-cellobiohydrolase. Figures 3 and 4 show Adsorption of cellulases on corn-cob residue was studied
the effects of β-glycosidase activity on the SSLF process. in independent experiments. The results are shown in
In the lag phase, a higher accumulated concentration of Figure 6. A Langmuir equation is able to correlate the
glucose was observed in Figure 4 because of higher equilibrium data.
activity of β-glycosidase. The other characteristics in
Figure 3 are similar to those in Figure 4. sat.
EC,ad ) EC,ad EC/(K + EC) (2)
Mathematical Model of the SSLF Process. In the
SSLF processes, both cellulose hydrolysis and lactic acid Hydrolysis of Cellulose into Cellubiose. The hy-
fermentation take place simultaneously. The process is drolysis of cellulose into cellubiose (G2) is catalyzed by
a multiphase system. At first, enzymes endo-β-1,4-glu- enzyme EC adsorbed on the active sites of lignocellulose
canase and exo-β-1,4-cellobiohydrolase (EC) are adsorbed (EC′), which takes place in the solid phase and is inhibited
on the corn-cob residue. The hydrolysis of the cellulose by cellubiose competitively. The reaction mechanism can
into cellubiose takes place in the solid phase by adsorbed be expressed as follows:
enzymes (EC′). And both the hydrolysis of cellubiose into
glucose by β-glucosidase (EB) and the lactic acid fermen- K1 k1
tation by L. delbrium occur in the liquid phase. Further, EC′ + C 9
7 8 EC′C 98 EC + G2 (3)
cellubiose and glucose are inhibitors for enzymes EC and
KG
enzyme EB, respectively. The whole picture of the process 2

can be simplified as shown in Figure 5. G2 + EC′ 798 EC′G2 (4)

Biotechnol. Prog., 1997, Vol. 13, No. 6 765

Because the rate of adsorption is much higher than Hydrolysis of Cellubiose into Glucose. The hy-
that of the reaction, and the concentration of the enzymes drolysis of cellubiose into glucose (G) is catalyzed by
is much lower than that of the substrate, therefore, the β-glycosidase (EB) in solution, which is inhibited by
overall reaction rate of eq 3 is independent of the product glucose; thus,
substrate concentration and is proportional to the con-
centration of enzyme EC′. Assuming EC′ is proportional K2 k2
G2 + EB 798 EBG2 98 EB + G (18)
to the total enzyme concentration being adsorbed, EC,ad,
the density of active sites, ns, and the surface area of KG
cellulose particle, A, then G + EB 798 EBG (19)
EC′ ) EC,adnSA (5) The reaction rate can be derived as

Assuming corn-cob residue is a spherical particle with
the same size, the total surface area is
A ) πd2N0 (6)
During the hydrolyzation process, the diameter of the
cellulose particle will be reduced; therefore, the number
of active sites should be a function of diameter
rG ) (1/YG/G2)k2EB0 exp(-kd2t)G2/(K2(1 + G/KG) + G2)
(20)
where the first-order deactivation of enzyme EB has been
included in eq 20.
Because the cellubiose is an intermediate, the concen-
tration is relative low, i.e., G2 , K2(1 + G/KG), and eq 20
can be simplified as

nS ) nS0πd2 ) nS0′C2/3 (7) exp(-kd2t)G2

rG ) B′EB0 (21)
1 + G/KG
where
where
nS0′ ) nS0 ( 3
FcπN0 ) n/3
(8)
B′ )
1 k2
(22)
YG/G2 K2
and the relationship between the concentration of cel-
lulose and diameter has been applied: Then the cellubiose accumulation rate can be calculated:

C ) (1/6)πd3FCN0 (9) dG2/dt ) rG2 - rG (23)

Combining eqs 7 and 9 and eliminating N0 and d by using Glucose Consumption by L. delbrium. In the
the initial cellulose concentration, C0, and diameter, d0, SSLF process, glucose produced from enzymatic hydroly-
then the surface area A can be expressed as sis will be further consumed by L. delbrium for cell
growth and lactic acid formation. The cell growth rate
can be described by Monod’s equation
A ) (6/FCd0)C01/3C2/3 (10)
dX/dt ) µmGX/(KS + G) (24)
The rate of reactions 3 and 4 can be written as
and the lactic acid formation is growth related:
rG2 ) k1EC′/(1 + G2/KG2) (11)
dP/dt ) R dX/dt (25)
or
Then, the glucose accumulation rate should be
4/3
rG2 ) BC (EC/(EC + K))(1/(1 + G2/KG2)) (12) dG/dt ) rG - β dX/dt (26)

where where

B ) (6nS0′/FCd0)C01/3EC,ad
sat.
(13) β ) 1/YX/G + R/YP/G (27)

If the deactivation rate of total enzyme EC can be
expressed as the first-order kinetics, i.e.,
EC,t ) EC0 exp(-kd1t)
and the enzyme concentration in solution, EC, is
EC ) EC,t - EC,ad
then, combining eqs 2, 14, and 15, we obtain
EC ) 1/2(H + (H2 + 4KEC0 exp(-kd1t))1/2)
where
sat.
H ) EC0 exp(-kd1t) - K - EC,ad
Parameter Determination. The parameters in Lang-
muir’s equation, eq 2, were correlated from experimental
sat.
data shown in Figure 6: EC,ad ) 0.0717 IU/gS; K )
(14) 1.413 IU/gS. The parameters B, B′, kd1, kd2, KG2, and KG
were simulated by use of eqs 12, 16, 21, and 23 from
experimental data of the enzymatic hydrolysis of corn-
cob residue: B ) 0.143; B′ ) 476; kd1 ) 1.24 × 10-4 min-1;
(15) kd2 ) 5.31 × 10-4 h/(g‚L); KG ) 0.13 g/L; KG2 ) 1.5 g/L.
The comparisons between experimental data and model
simulation are shown in Figure 1.
It is difficult to determine cell density, X, in the SSLF
process because the cells will be adsorbed on the ligno-
(16)
cellulosic materials. The parameters in the Monod
equation were obtained from literature (Roger et al.,
1978): µm ) 0.296 1/h and Ks ) 0.545 g/L. Only the
parameter β was correlated from the SSLF experimental
(17) data: β ) 6.87.
766 Biotechnol. Prog., 1997, Vol. 13, No. 6

Table 5. Comparisons between SSLF and Two-Step EC,ad parameter in Langmuir’s equation, FPIU/gS
Processes (E0 ) 5.0 FPIU/gS, C0 ) 43g/L) ECC cellulase-cellulose complex concentration, (mg/
period G2 G P Y* a Y mL)‚(FPIU/mL)
(h) (g/L) (g/L) (g/L) (%) (%) Et total enzyme concentration, FPIU/mL
two-step process G glucose concentration, g/L
saccharification 72 8.3 22.7 79.0 G2 cellubiose concentration, g/L
fermentation 48 8.3 1.35 20.84 71.0 59.0
K equilibrium constant for cellulase-cellulose com-
SSLF 72 0.25 0.46 33.97 79.0 79.0
plex, FPIU/mL
a Y* ) cellulose utilization ratio. KS substrate limitation constant, g/L
KG inhibitory constant for cellobiose hydrolysis, g/L
Comparisons between Experimental Data and K G2 inhibitory constant for cellulose hydrolysis, g/L
Model Calculation. The comparisons between the K1, K2 equilibrium constant
experimental data and the model calculation of the SSLF
kd1 deactivation constant of cellulase, 1/h
process are shown in Figures 3 and 4. It is obvious that
the proposed model is capable of describing the SSLF kdB deactivation constant of β-glucosidase, 1/h
process. During the SSLF process, the concentration of k1, k2 rate constant, 1/h
cellubiose is almost nil; therefore, the inhibition of N number of particles, 1/L
cellubiose on enzyme is negligible. And the accumulation ns density of active site, mmol/m2
of glucose is observed and predicted by the model in the ns0, ns0 site density constant, mmol/m2
early stage of the SSLF processes. Because the cells are P lactic acid concentration, g/L
still in the lag phase and at low density, the rate of rG reaction rate of glucose formation from cellobiose,
glucose formation by enzymatic hydrolysis is higher than g/(L‚h)
the rate of glucose consumption by L. delbrium, especially rG2 reaction rate of cellubiose formation from cel-
when β-glucosidase activity is high (see Figure 4). After lulose, g/(L‚h)
that, L. delbrium was in exponential growth phase and t time, h
glucose was consumed rapidly. The concentration of
X biomass concentration, g/L
glucose was kept in a very low level, which meant the
inhibition caused by glucose was negligible also. Y enzymatic hydrolyzation yield, %
There are deviations between model prediction and Y/ yield constant
experimental data as seen in Figures 3 and 4. The Greek Symbols
reason is the oversimplification in the model. For
example, L. delbrium will be adsorbed on the lignocel- R constant
lulosic materials, which can cause the mass transfer β constant
limitation to lower the production rate of lactic acid. Fc particle density, g/m3
Comparisons between the SSLF and Two-Step µm maximum specific growth rate, 1/h
Processes. The comparisons between the SSLF and
Subscript
two-step processes are shown in Table 5. It is obvious
that the SSLF process is higher in productivity as well 0 t)0
as lactic acid yield than the two-step process. In the two-
step process, the lactic acid fermentation is carried out
after enzymatic saccharification of the corn-cob residue. Literature Cited
During the saccharification stage, the ratio of cellulose
being hydrolyzed is only 79% because of product inhibi- Abe, S.; Takagi, M. Simultaneous saccharification and fermen-
tation of cellulose to lactic acid. Biotechnol. Bioeng. 1991, 37,
tion. In the fermentation step, the cellubiose cannot be
93-96.
metabolized by β-glycosidase; the cellulose utilization
Barker, S. B.; Summerson, W. H. The colorimetric determination
ratio is also low. The overall yield of lactic acid is only of lactic acid in biological material. J. Biol. Chem. 1941, 138,
59%, compared to 79% in the SSLF process. The total 538-554.
period of the SSLF process (72 h) is much shorter than Beltrame, P. L.; Carniti, P.; Focher, B. Enzymatic hydrolysis of
that of the two-step method (total of 120 h). cellulosic materials: A kinetic study. Biotechnol. Bioeng.
1984, 26, 1233-1238.
Conclusions Bergmeyer, H. Methods of Enzymatic Analysis; Verlag Che-
mie: Weinheim, Berlin, 1974; p 1025.
1. Enzymatic saccharification of corn-cob residue is
Dekker, R. F. H. Kinetic, inhibition, and stability properties of
perfectly compatible with lactic acid fermentation in a
a commercial β-D-glucosidase(cellobiase) preparation from
SSLF process with high lactic acid productivity and yield. Aspergillus niger and its suitability in hydrolysis of lignocel-
2. The proposed kinetic model is capable of predicting lulose. Biotechnol. Bioeng. 1986, 28, 1438-1442.
the SSLF process. Enari, T.-M. Microbial cellulases. In Microbial Enzymes and
This work is supported by the International Coopera- Biotechnology; Fogarty, Ed.; Applied Science: London, 1983;
tive Research Fund from Science and Technology Com- p 183-223.
mittee of Zhejiang Province, PRC. Ghose, T. K. Measurements of cellulase activities. Pure Appl.
Chem. 1987, 59, 257-268.
Notation Goncalves, L. M. D.; Xavier, A. M. R. B.; Almeida, J. S.;
Carrondo, M. J. T. Concomitant substrate and product
A equivalent spherical interfacial area, m2/mL inhibition kinetics in lactic acid production. Enzyme Microb.
B proportionality factor Technol. 1991, 13, 314-319.
C cellulose concentration, g/L Jing, Q. R.; Zhang, J. M.; Xu, J. Processes of Organic Acid
d mean particle diameter, m Frementation, 2nd ed.; Light Industry Press: Beijing, 1995;
pp 339-378 (in Chinese).
EB β-glucosidase activity, BU/mL
Lee,Y. H.; Fan, L. T. Properties and mode of action of cellulase.
EC enzyme concentration in solution, FPIU/mL In Advances in Biochemical Engineering; Fiechter, A., Ed.;
sat. parameter in Langmuir’s equation, FPIU/gS
EC,ad Springer: Berlin, 1980; Vol. 17, p 101.
Biotechnol. Prog., 1997, Vol. 13, No. 6
Lipinsky, E. S.; Sinclair, R. G. Is lactic acid a commodity
chemical? Chem. Eng. Progr. 1986, 82 (Aug), 26.
Philippidis, G. P.; Smith, T. K.; Wyman, C. E. Study of the
enzymatic hydrolysis of cellulose for production of fuel ethanol
by the simultaneous saccharification and fermentation pro-
cess Biotechnol. Prog. 1993, 41, 846-853.
Roger, P. L.; Bramall, L.; McDonald, I. J. Kinetic analysis of
batch and continuous culture of Streptococcus cremoris HP.
Can. J. Microb. 1978, 24, 372-380.
South, C. R.; Lynd, L. E. Analysis of conversion of particulate
biomass to ethanol in continuous solids retaining and biore-
actors. Appl. Biochem. Biotechnol. 1994, 45/46, 467-481.
Spindler, D. D.; Wyman, C. E.; Grohmann, K. Evaluation of
thermotolerant yeasts in controlled simultaneous sacchari-
fications and fermentations of cellulose to ethanol. Biotechnol.
Bioeng. 1989, 34, 189-195.
767
Takagi, M. Inhibition of cellulose by fermentation products.
Biotechnol. Bioeng. 1984, 26, 1506-1507.
Xu, J. P.; Liu, J. S.; Kong, W.; Zhang, S. J. SCP production from
straw by mixed microbs culture. Microbiology 1995, 22, 222-
227 (in Chinese).
You, X. Production and Application of Xylitol; Light Industry
Press: Beijing, 1984 (in Chinese).
Accepted September 11, 1997.X
BP970100H
X Abstract published in Advance ACS Abstracts, November 1,
1997.