• Applied Biological Chemistry
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• v.24 no.2
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• pp.85-93
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• 1981

For the utilization of natural cellulosic materials by microorganisms, a potent cellulase-producing microorganism was isolated and identified as Trichoderma spp. Rice straw used as a substrate in this study was preliminarily treated with chemical solvents and/or additionally treated with acids and by heat, and then examined with the cellulase produced by the organism. Better results in sugar production by decomposing the straw cellulose were obtained, when the cellulase was produced by cultivating the organism in the selection medium, pH 5.0, for 5 days, and when the pretreated straw substrate was additionally treated with 0.1% $H_2SO_4$ sulfuric acid at $120^{\circ}C$ for 1 hour. The enzyme production was increased by about 20%, when 0.5% urea 0.5% phosphate, 0.1% meat extract, or 5% orange peel was added into the culture medium. For the practical purposes, the sugar production from the rice straw by the cellulase-producing microorganism can be improved by extending the reaction time of the enzyme up to 24 hr or longer.

• Journal of Microbiology and Biotechnology
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• v.17 no.11
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• pp.1782-1788
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• 2007

Cellulosomes in Clostridium cellulovorans are assembled by the interaction between the repeated cohesin domains of a scaffolding protein (CbpA) and the dockerin domain of enzyme components. In this study, we determined the synergistic effects on cellulosic and hemicellulosic substrates by three different recombinant mini-cellulosomes containing either endoglucanase EngB or endoxylanase XynA bound to mini-CbpA with one cohesin domain (mini-CbpAl), two cohesins (mini-CbpA12), or four cohesins (mini-CbpAl234). The assembly of EngB or XynA with mini-CbpA increased the activity against carboxymethyl cellulose, acid-swollen cellulose, Avicel, xylan, and com fiber 1.1-1.8-fold compared with that for the corresponding enzyme alone. A most distinct improvement was shown with com fiber, a natural substrate containing xylan, arabinan, and cellulose. However, there was little difference in activity between the three different mini-cellulosomes when the cellulosomal enzyme concentration was held constant regardless of the copy number of cohesins in the cellulosome. A synergistic effect was observed when the enzyme concentration was increased to be proportional to the number of cohesins in the mini-cellulosome. The highest degree of synergy was observed with mini-CbpAl234 (1.8-fold) and then mini-CbpAl2 (1.3-fold), and the lowest synergy was observed with mini-CbpAl (1.2-fold) when Avicel was used as the substrate. As the copy number of cohesin was increased, there was more synergy. These results indicate that the clustering effect (physical enzyme proximity) of the enzyme within the mini-cellulosome is one of the important factors for efficient degradation of plant cell walls.

• Journal of the Korean Wood Science and Technology
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• v.42 no.2
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• pp.172-182
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• 2014

Extraction is a necessary element in the bioconversion of lignocellulosics to fuels and chemicals. Although various forms of chemical pretreatment of cellulosic materials have been proposed, their effectiveness varies depending on the treatment conditions and substrate. In this study, mixed hardwood (MH) and loblolly pine (LP) were pretreated with dilute acid in a 100 mL accelerated solvent extraction (ASE) at the predetermined optimal conditions: temperature: $170^{\circ}C$, acid concentration: 0.5% (w/v), and reaction time: 2~64 min. This method was highly effective for extracting the hemicellulose fraction. Total xmg (defined as the sum of xylose, mannose, and galactose) can be extracted from milled MH and LP through pressurized dilute acid treatment in maximum yields of 12.6 g/L and 15.3 g/L, respectively, representing 60.5% and 70.4% of the maximum possible yields, respectively. The crystallinity index increased upon pretreatment, reflecting the removal of the amorphous portion of biomass. The crystalline structure of the cellulose in the biomass, however, was not changed by the ASE extraction process.

• Microbiology and Biotechnology Letters
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• v.4 no.3
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• pp.99-104
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• 1976

Experiments were carried out to establish the effects of acids in neutralization after alkaline treatment of rice straw, with which cellulosic single cell protein can be produced by cellulose utilizing bacteria, Cellulomonas flavigena KIST 321, previously isolated by authors. Following results were obtained. 1. Rice straw as carbon source was pretreated with 10 volumes of 1 normality of NH$_4$OH or NaOH(NaOH/substrate：40％, and then washed with water or neutralized with H$_3$PO$_4$, H$_2$SO$_4$, HCl and CH$_3$COOH. Among the above mentioned methods, neutralization with H$_3$PO$_4$after alkaline treatment was proved to be the most effective on its digestibility and SCP production. Dry cell 12.28g/$\ell$ and 78％ digestibility were obtained. 2. When rice straw was treated with NaOH solution, the result suggested that the productibity of cell-mass was attained on treatment of rice straw with 6％ of NaOH (NaOH/substrate ratio) for 15～24hrs at room temperature. 3. When rice straw was treated with NaOH, a volume of water to substrate is adequate by two or three fold and the amount of NaOH can be economized up to 5％ for the weight of rice straw. 4. The steaming of rice straw at 121$^{\circ}C$ for 30min. in alkaline treatment of rice straw gave the similiar effectiveness to that at room temperature for 15～24hrs. and accelerated the sterilization of the substrate. 5. Finally, the level of inorganic phosphate in a medium was investigated. 11.2g of dry cell was produced at the concentration of 0.2％, phosphate (phosphorous level 0.04％) in medium even though treated rice straw was neutralized with HCI instead of H$_3$PO$_4$, and 12.2g/$\ell$ at the concentration of 0.3％ phosphate (phosphorous 0.04％) on neutralization with H$_2$SO$_4$.

• Microbiology and Biotechnology Letters
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• v.16 no.2
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• pp.150-155
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• 1988

In order to produce L-lysine from cellulosic substrates by the intergeneric protoplast fusion between cellulolytic bacteria, Cellulomonas flavigena KFCC31221 and amino acid producing bacteria, Brevibacterium flavum ATCC14067, Corynebacteriurn glutamicum ATCC13032, conditions for protoplast formation and regeneration of these strains were investigated. After the strains were mutated with 500$\mu\textrm{g}$/$m\ell$ N-methyl-N'-nitro N-nitrosoguanidine for 30 min and the mutants were enriched by treating 300$\mu\textrm{g}$/$m\ell$ penicillin-G for 2 hrs, B. flavum Hse- Str$^{r}$ , C. glutamicum Met$^{-}$Thr$^{-}$ Rif$^{r}$ and Cellulomonas flavigena Thr$^{-}$Val$^{-}$Kan$^{r}$ were isolated. The rate of protoplast formation ranged from 95 to 98% when strains were treated at the concentration of 500$\mu\textrm{g}$/$m\ell$ of lysozyme, pH 6.5, 33$^{\circ}C$, for 6 hrs. in Tris- malate buffer supplemented with 0.4M sucrose as osmotic stabilizer. Approximately 30-33% protoplast was regenerated on the regeneration complete medium(RCM) containing 1.5% agar and 0.5M sodium succinate overlaid with the same medium except 0.7% agar.

• Applied Biological Chemistry
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• v.27
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• pp.29-46
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• 1984

To utilize several species of hard wood as raw materials of feed products, fermentation characteristics of cellulosic substrates to single cell protein was investigated, and results were summarized as follows. Among the microorganisms investigated, Tricoderma viride was selected as one of the most cellulolytic. Mixed culture of fungi did not show a synergistic effect on cellulose degradation. When the fungi were cultured at $28^{\circ}C$ for 7 days in a medium containing wheat bran 25 g, cellulose 0.25 g, proteose peptone 0.025 g and tween 800.025 g, cellulotic activities on carboxy methyl cellulose and filter paper reached maximum at 12 hr. The alkali treatment resulted in increased degradation of substrate from 13 to 18% when treated with enzymes for 12h, and reducing sugar formation increased with decreased size of substrates. Glucose was a very good feedback inhibitor of the enzyme from T.viride than that of xylose. When the substrate was rehydrolyzed, hydrolysis rate was 31% to reducing sugars within 12 hr. Quantative anlysis with HPLC showed the ratio of glucose to xylose in sugar syrups as 1.77 to 1. For the purpose of producing cellulosic-single cell protein from the sawdust of mulberry tree, 15 strains of xylose-assimilating yeast were isolated from 42 samples of rotten woods and compost soils and examined for their ability to utilize xylose. Then three strains were selected by their strong xylose-assimilating activities. The cultivative condition, the growth characteristics, and protein and nucleic acid productivities of three strains were investigated. The results obtained were, 1. Wood hydrolysate of mulberry tree was assimilated by 5 strains of CHS-2, CHS-3, ST-40, CHS-12 and CHS-13. 2. The optimum initial pH and temperature for the growth of strain CHS-13 were 4.4 and $30^{\circ}C$. 3. The specific growth rate of strain CHS-13 was $0.23h^{-1}$ and generation time was 3.01 hrs at the optimum condition. 4. CHS-13 strain assimilated 81 % of sugar in wood hydrolysate. 5. CHS-13 strain was identified as Candida guilliermondii var. guilliermondii 6. When the CHS-13 strain was cultured in the wood hydrolysate containing yeast extract, L-protein content was increased with yeast extract concentration. 7. The L-protein and nucleic acid yields from wood hydrolysate were 0.73 mg/ml and $4.92{\times}10^{-2}\;mg/ml$ respectively. 8. An optimal nucleic acid content of CHS-13 strain was observed in the medium containing 0.2% of yeast extract.

• Microbiology and Biotechnology Letters
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• v.16 no.3
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• pp.213-218
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• 1988

Studies were made to optimize the simultaneous saccharification and fermentation (SSF) of rice straw to produce butanol using Clostridium acetobutylicum KCTC 1037 and a cellulolytic enzyme preparation from Trichoderma viride. The fermentation was inhibited when the liquid enzyme preparation from Novo was used, whilst a successful fermentation was achieved in the SSF using the enzyme manufactured by Pacific Chemical Co. The minimum cellulase concentration for the successful fermentation of pure cellulose was found to be 4 IU/g of substrate used. Alkaline treatment was better method for the fermentation of rice straw by the system. SSF using 25％ alkaline treated rice straw produced 150 mM butanol, 90 mM acetone. On the other hand, fermentation of ball milled rice straw was mainly acidogenic producing 98 mM acetate and 64 mM butyrate with less than 20 mM butanol. These results show that rice straw contains (a) specific inhibitor(s) for solventogenesis which is destroyed or soluble in alkali.

• The Korean Journal of Food And Nutrition
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• v.10 no.3
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• pp.344-349
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• 1997

A strain of Bacillus sp. DSNC 101, isolated from soil, produced up to 305.0 units/ml of xylanase when grown on te medium containing 2.0% xylan, 2.0% yeast extract and 0.4% K2HPO4. The strain produced xylanase in the presence of xylan, soluble starch, rice straw, Avicel, maltose, and lactose as a sole carbon source, but the enzyme was not synthesized in the presence of xylose, glucose or arabinose. The crude xylanase preparation did not show hydrolytic activity towards cellulosic substrates and PNPX, a chromogenic substrate for $\beta$-xylosidase. The temperature and pH optima for the xylanase production were 4$0^{\circ}C$ and 8.0, respectively. Xylanase synthesis was repressed by glucose, but not by xylose. The hydrolysis products of xylan catalyzed with the culture filtrate were xylooligosaccharides such as xylobiose and xylotriose but xylose was not detected by tin layer chromatography.

• The Korean Journal of Mycology
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• v.17 no.2
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• pp.57-65
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• 1989

The effect of various cellulosic and hemicellulosic substrates on the induction of cellulase and xylanase complexes in Aapergillus nidulans was investigated. The most efficient substrates for the induction of cellulase and xylanase complexes were carboxymethylcellulose for endoglucanase, cellobiose for ${\beta}-glucosidase$, and xylan for endoxylanase and ${\beta}-xylosidase$, respectively. However, the mixtures of these substrates, especially CMC-xylan and CMC-xylan-laminarin mixture, were much more effective not only for the enhancement of the biosynthesis of all the cellulase and xylanase complexes but also for the balanced production of these enzyme components than individual substrate. The polyacrylamide gel electrophoresis followed by activity staining showed the variation in the patterns and relative intensity of ${\beta}-glucosidase$, endoglucanase and endoxylanase components in individual enzyme preparations from A. nidulans cultures grown on different substrates. These results suggest that the biosynthesis is of cellulase and xylanase systems in A. nidulans is regulated in coordination at the level of induction.

• KSBB Journal
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• v.31 no.1
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• pp.73-78
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• 2016

The aim of this study attempted to verify the possibility of bioethanol production using wasted medium density fiberboard (wMDF). In order to produce bioethanol from wood cellulosic materials must be carried out the process of pretreatment, saccharification, fermentation and distillation. First, the wMDF was pretreated using sodium chlorite and pretreated wMDF was prepared to 8% slurry and then slurry was saccharified with the commercial enzyme (Cellic CTec3). The fermentable sugar and pH of saccharified substrate were about 5.5% glucose and 4.4, respectively. Herein we compared the results of ethanol yield according to the nutrients added or without addition to increase ethanol yield. Ethanol fermentation was finished in about 24 hours, but it was delayed in experimental group without nutrients. Ethanol content and fermentation ratio of the final fermented mash prepared by utilizing jar fermenter was 25.40 g/L and 86.64%, respectively. At this time, the maximum ethanol productivity was confirmed as 1.78 g/Lh (ethanol content 21.38 g/L, 12 h), and the overall ethanol productivity was 1.05 g/Lh (ethanol content 25.27 g/L, 24 h). Using fermented liquid we could produced bioethanol 95.37% by continuous distillator packed with copper element in laboratory scale. These results show that wMDF has a potential valuable for bioethanol production.