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3.5.1 Outline
3.5.2 Pre-treatment of cellulosic biomass
3.5.3 Cellulase production
3.5.4 Saccharification of biomass
3.5.5 Enzyme recovery from biomass
3.5.6 Concentration of sugar solutions
3.5.7 Alcohol fermentation
3.5.8 Alcohol recovery
Various aspects of alcohol fermentation from cellulosic biomass have been discussed thus
far. A number of problems still remain to be resolved prior to industrial-scale production of
fuel alcohol from cellulosic biomass. As mentioned in the introduction to this chapter,
subsequent to our studies on basic and elemental techniques, an integrated pilot plant for
the production of alcohol from biomass, was constructed in our laboratory, in order to
demonstrate individual processes. Construction of the plant commenced in 1983, and
continued in a step-wise manner for 5 years. The final plant was capable of treating 720 kg
of raw material per day with the production of 150 to 200 liters of dehydrated fuel alcohol.
A process flow diagram of the pilot plant is given in Fig. 3-20, while Fig. 3-21 shows a plan
of the plant.
Figure 3.17 - Effect of alcohol concentration on fermentation rates of immobilized
continuous fermentation processes
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3.5.1 Outline
Bagasse, rice straw, and beech wood chips, the compositions of which are shown in Table
3-6, were used as cellulosic biomass in the pilot plant. These materials vary in
composition, and are thus presumed to differ in three-dimensional structure. Cellulose can
either be saccharified by an acid process, or by an enzymatic process using cellulase. For
the pilot plant, the enzymatic process was adopted in view of problems associated with the
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acid process, i.e., the necessity for the use of acid-resistant equipment, technical
difficulties relating to the recovery of acid, and the formation of by-products due to
excessive decomposition. At the outset, cellulase of a high titer and balanced enzymatic
activity was unavailable, which meant that a long period of time was required for
saccharification. However, by employing a variety of screening and mutational techniques,
a high titer cellulase that could be produced at a low cost was developed. Saccharification
from both low- and high-concentration biomass was investigated, and an appraisal of the
most suitable types of reactor was made. Owing to the high cost of cellulase, its recovery
and re-use were investigated using UF techniques. With the objective of improving the
efficiency of alcohol fermentation and thus saving energy, the use of RO as a means of
concentrating the sugar solutions was investigated. Alcohol fermentation was conducted
by the immobilized flash method using a bioreactor in combination with the flash method
for the purpose of accelerating alcohol production while minimizing the influence of alcohol
generated on yeast cells. Two methods were used to evaluate the recovery of alcohol: the
conventional azeotropic distillation method, and a combination of the super critical fluid
extraction (SCFE) and pervaporation methods. In addition, the plant was evaluated as a
total system using waste water, which mainly consisted of lignin waste water derived from
the pre-treatment step and alcohol fermentation waste water. This waste water was treated
by the methane fermentation method using a cell concentration apparatus fitted with a
membrane. Low-concentration alcohol leaking from the SCFE-pervaporation, and
immobilized flash fermentation steps was recovered using RO. The processing steps
employed in the plant are described individually in the following Sections.
Figure 3.19 - Time profile of fermentation using an immobilized flash fermentation
system
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Bacteria represented by the genus Cellulomonas, and yeast strains including those of the
genus Trichosporon were used for cellulase production. In general, the production of
cellulase by fungi was preferred. As discussed in Sections 3.2.2 and 3.2.3, various
microorganisms obtained domestically as well as from abroad, were screened, and
Trichoderma reesei QM-9414 isolated by the U.S. Army Natick Laboratory was selected
as the best strain. Mutants with enhanced enzymatic titer or with specific desirable
characteristics were obtained by mutation using QM-9414 as a parent strain.
The semi-batch culture method produced a high-titer cellulase. A culture method
incorporating pH shifting was employed to enhance p-glucosidase activity in Trichoderma
microorganisms. In addition to scale-up experiments, cheese whey and biomass (bagasse,
rice straw, etc.) were investigated as potential inexpensive carbon sources, while soybean
meal and residual fermentation cells, were investigated as potential inexpensive nitrogen
sources. On the basis of results obtained, a scale-up experiment was carried out in a 1-kL
tank using the semi-batch culture method with 10% Avicel (crystalline cellulose) as a
carbon source and soybean meal as a nitrogen source, with pH control by the shifting
method. The time course of cellulase production by T. reesei CDU-11 under these
conditions is shown in Fig. 3-22. High-titer cellulase activities, i.e., 720 U/ml as CMCase,
33 U/ml as FPU, 47 U/ml as Avicelase, and 31 U/ml as p-glucosidase, were obtained.
Soluble protein resulting exceeded 5 %.
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SCFE-Pervaporation
98.8
0.15
0.16
0.21
0.45
Azeotropic distillation
99.5
0.05
0.03
0.003
0.33
H2O: wt. %
Others: v/v%
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