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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
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 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 byproducts 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
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 %.
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 SCFE-Pervaporation Azeotropic distillation H2O: wt. % 98.8 99.5 Composition of Sample (%) EtOH BuOHs AmyO Hs CH3CHO H2O 0.15 0.05 0.16 0.03 Others: v/v% 0.21 0.003 0.45 0.33