3.5 Alcohol Production Using An Integrated Pilot Plant

3.
5 Alcohol production using an integrated pilot plant
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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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Figure 3.18 - Schematic of an immobilized flash system
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
Figure 3.20 - Flow diagram outlining processes in an integrated bench plant
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Figure 3.21 - Schematic of the layout of an integrated bench plant
3.5.2 Pre-treatment of cellulosic biomass

Pre-treatment of cellulosic biomass was conducted as described in Section 3.3.1.
Bagasse and rice straw were crudely crushed and pre-treated with alkali at 90 to 120C for
1 to 2 hours in a double screw-type counter-current extractor. The optimum ratio by weight
of sodium hydroxide to biomass was 0.08 to 0.1. In the case of beech wood chips, since
saccharification did not proceed with sodium hydroxide treatment, steam explosion was
found to be effective. Steam explosion combined with treatment by alkali and salts
produced good results.
3.5.3 Cellulase production
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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 %.
3.5.4 Saccharification of biomass

High-concentration saccharification was conducted by the batch method. An example
using bagasse as the biomass source is shown in Fig. 3-12. Low-concentration
saccharification was performed by the continuous method, the volume of biomass being
adjusted to between 3 and 6%. A saccharification rate of 90 to 95%, was obtained after 5
to 7 hours of reaction, with only a small amount of unreacted biomass residue remaining
after the reaction. Beech chips treated by steam explosion (Temp: 220C2 pressure
retention time: 5 min) were also subjected to the saccharification reaction for 24 hours,
resulting in a saccharification rate of approximately 75 %. Table 3-7 summarizes cellulase
requirements, for the saccharification of biomass, using cellulases prepared in our
laboratory.
Figure 3.22 - Time course of high titer cellulase production by T. reesei CDU-11
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3.5.5 Enzyme recovery from biomass

As explained in Section 3.2.1, cellulase encompasses a group of enzymes having different
molecular weights. A UF membrane capable of fractionating molecular weights ranging
between 10,000 and 20,000 Dalton, was therefore used to recover cellulase for re-use.
While enzyme recovery from low-concentration saccharification solutions was
approximately 90%, that from high-concentration saccharification solutions was
approximately 75 to 80% owing to cellulase adsorption by saccharification residues.
Enzyme recovery in the latter case could be markedly improved by re-slurrying the residue
to extract the enzymes. In the pilot plant, tubular-type ultrafiltration equipment with a
polystyrene membrane was used.
3.5.6 Concentration of sugar solutions

In order to improve the efficiency of alcohol fermentation, UF-filtered sugar solutions which
were also free of enzyme were passed through a polyamide, spiral-type RO membrane,
enabling them to be concentrated from between 3 and 15% to a final concentration of
approximately 30%. Using this loose RO system, more than 95% of the sugar was
recovered over an extended period.
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3.5.7 Alcohol fermentation

The immobilized flash method was used for alcohol fermentation in the integrated
pilot-scale plant. Results are shown in Fig. 3-23. Using this method, it was possible to
maintain the alcohol concentration at approximately 5%, while elevating the sugar
concentration of the raw material to as high as 30%. Table 3-9 compares fermentation
methodologies used in alcohol production. Three immobilized yeast fermentors, each
provided with a flash distillation tank, were connected in series in the pilot plant.
In order to efficiently convert xylose contained in the saccharified solutions of biomass,
yeast strains were screened and Kluyveromyces cellobiovorus, a strain capable of
assimilating cellobiose and producing ethanol by fermentation was newly isolated. This
strain was however of poor alcohol resistance, as are other known xylose-assimilating
yeast strains. The simultaneous use of the flash method was effective in minimizing the
influence of alcohol and efficiently inducing the activity of this strain.
3.5.8 Alcohol recovery

Methods used for recovering alcohol included the distillation method, and the SCFE and
osmotic vaporization methods.
In the distillation method, dehydrated ethanol was obtained by ternary azeotropic
distillation, with benzene as a solvent. A pressure-type concentrator was employed, with
steam at the top of the concentrator as an energy source for the ethanol separation
column. Using this methodology, energy savings of 30% over that of the conventional
method were achieved.
Figure 3.23 - Time profile of ethanol production, using the immobilized yeast and
flash system
In the SCFE, ethanol obtained from the flash distillation step, ranging in concentration from
20 to 30% was brought into contact with supercritical carbon dioxide gas, resulting in 90 %
recovery of 85 to 95% concentrated ethanol solutions. By optimizing the extraction
pressure and temperature and, at the same time, providing a mist separator at the top of
the extractor, ethanol having a composition approaching that of the azeotrope was
recoverable. Impurities in the culture broth exhibited the same behavior as that of the
ethanol. Studies on osmotic vaporization membrane methods for dehydration, resulted in
alcohol concentrations exceeding 99.5 % at a recovery rate of at least 95 %. Analytical
values for fuel alcohol obtained by the ternary azeotropic method, and by supercritical
carbon dioxide extraction in combination with an osmotic vaporization membrane are
shown in Table 3-10. In both methods, the water content of alcohol recovered, was less
than 0.5%, indicating that this alcohol may be feasibly used as fuel ethanol. Dehydrated
ethanol produced from bagasse was mixed with gasoline and its performance as a fuel for
automobiles was evaluated by Cosmo Oil Co., Ltd. Results showed no substantial
difference from ordinary reagent ethanol, and the ethanol was deemed suitable for
practical use.
Table 3-10 Composition of Dehydrated Ethanol
Method of Dehydration
Composition of Sample (%)

EtOH BuOHs AmyO Hs CH3CHO H2O
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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http://www.fao.org/docrep/w7241e/w7241e0b.htm#3.5 alcohol productio...

Produced by: Agriculture and