• Journal of Applied Biological Chemistry
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• v.61 no.1
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• pp.59-67
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• 2018

This study was conducted to isolate the cellulolytic bacteria able to grow on LB- Carboxymethyl cellulose (CMC) agar trypan blue medium from the mixed forest and Larix leptolepis stands. Three bacterial strains with high activity against both CMC and xylan were isolated. Both API kit test and 16S rRNA gene sequence analysis revealed that the three different isolates belong to the gene Bacillus. Therefore, the isolates named as Bacillus sp. EFL1, Bacillus sp. EFL2, and Bacillus sp. EFP3. The optimum growth temperature of Bacillus sp. EFL1, EFL2, and EFP3 were $37^{\circ}C$. The optimum temperature for CMCase and xylanase from Bacillus sp. EFL1 were $50^{\circ}C$. The optimum pH of Bacillus sp. EFL1 xylanase was pH 5.0 but the optimum pH of CMCase from Bacillus sp. EFL1 was pH 6.0. The optimum temperature of CMCase and xylanase from Bacillus sp. EFL2 was $60^{\circ}C$, respectively. The optimum pH of CMCase of Bacillus sp. EFL2 was 5.0, whereas xylanase showed high activity at pH 3.0-9.0. The optimum temperature for CMCase and xylanase of Bacillus sp. EFP3 was $50^{\circ}C$. The optimum pH for CMCase and xylanse was 5.0 and 4.0, respectively. CMCases from Bacillus sp. EFL1, EFL2, and EFP3 were thermally unstable. Although xylanase from Bacillus sp. EFL1 and EFP3 showed to be thermally unstable, xylanase from Bacillus sp. EFL2 showed to be thermally stable. Therefore, Bacillus sp. EFL2 has great potential for animal feed, biofuels, and food industry applications.

• Journal of Microbiology
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• v.34 no.4
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• pp.305-310
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• 1996

A microorganism producing carboxymethyl-cellulase (CMCase) was isolated from 300 soil and compost samples. The isolate was identified as Bacillus sp. by $Biolog^{TM}$ test and fatty acid analysis, and named as Bacillus sp. KD1014. The isolate could degrade, in addition to CMC, various kinds of polysaccharides such as levan, xylan, starch, and filter paper but hardly degrade microcrystalline Avicel. The optimum growth and CMCase production of the isolate was observed between 16-and 25 hr-culture at 45$^{\circ}C$ and pH 5.0. The maximum CMCase activity was observed at pH 4.5 and 6$0^{\circ}C$. The CMCase was found to bind to Avicel. The CMCase was internally cleaved as growth continued. When crude supernatant was used for activity staining, three major bands were detected on a native gel, however, only one major band was detected on a denaturating gel after removal of the detergent.

• Microbiology and Biotechnology Letters
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• v.25 no.6
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• pp.546-551
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• 1997

A bacterium producing the extracellular cellulases was isolated from soil and has been identified as Bacillus sp. The isolate, named Bacillus sp. 79-23, was shown to be very similar to B. subtilis on the basis of its biochemical properties. The carboxymethyl cellulase (CMCase) of culture supernatant was most active at 60$\circ$C and pH 6.0, and retained 90% of its maximum activity at pH 7.0. The additional carbon sources affected the CMCase productivity than nitrogen sources in the culture medium. The carbon sources including wheat bran, rice straw, maltose and glucose increased the enzyme productivty. Especially, the maximum CMCase production was 5.2 units/ml in LB medium supplemented with 3% (w/v) wheat bran, which was 13-folds more than that in LB medium. It was found that the enzyme production was in association with the growth of Bacillius sp. 79-23. But, whean bran did not affect the growth of isolate, suggesting that increasement of CMCase production was owing to the induction of CMCase biosynthesis by wheat bran. In addition, both water-soluble and insoluble components of wheat bran was involved in induction of CMCase biosynthesis.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.524.2-524
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• 1986

The Bacillus CMCase gene we have previously cloned in E. coli is contained in the 3.2 Kb chromosomal insert of the 7.5 Kb pBSl plasmid. We have also found that the CMCase produced by this gene has molecular weight of about 32,000 suggesting that the CMCase coding region lies on about 0.3 Kb fragment. The present report deals with a series of subclonings to localize more precisely the region coding for the CMCase production.

• Journal of Mushroom
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• v.14 no.4
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• pp.244-248
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• 2016

In order to isolate thermophilic bacteria with high activity of CMCase and xylanase, spent mushroom substrates was collected from an oyster mushroom cultivation farm in Jinju, Gyeongnam, Korea. Among the isolates, one strain designated as YJ09 was selected by agar diffusion method. The isolate YJ09 was identified as a member of Bacillus licheniformis based on biochemical characteristics using Bacillus ID kit and MicroLog system. Comparative 16S rDNA sequence analysis showed that isolate YJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus licheniformis with sequence similarity of 98.9%. Based on its physiological properties, biochemical characteristics and phylogenetic distinctiveness, the isolate YJ09 was classified as Bacillus licheniformis. The CMCase and xylanase activity of B. licheniformis YJ09 was slightly increased corresponding to the bacterial population from exponential phase to stationary phase in the growth curve of B. licheniformis YJ09.

• Microbiology and Biotechnology Letters
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• v.31 no.4
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• pp.383-388
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• 2003

A bacterium producing the extracellular xylanase and CMCase was isolated from soil and has been identified as Bacillus sp. The isolate, named Bacillus sp. A-7, was shown to be very similar to Bacillus licheniformis on the basis of its biochemical and physiological properties. The maximum xylanase and CMCase production were obtained when 2.0% (w/v) glucose and 0.3% (w/v) yeast extract were used as carbon source and nitrogen source, respectively. The best mineral conditions for xylanase and CMCase production were 0.1%(w/v) $CaC1_2$. Among the various feedstuffs, 1.0%(w/v) soybean meal was selected for the best xylanase and CMCase production.

• Journal of Life Science
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• v.21 no.8
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• pp.1083-1093
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• 2011

A microorganism utilizing rice hulls as a substrate for the production of carboxymethylcellulase (CMCase) was isolated from seawater and identified as Bacillus lincheniformis by analyses of its 16S rDNA sequences. The optimal carbon and nitrogen sources for production of CMCase were found to be rice hulls and ammonium nitrate. The optimal conditions for cell growth and the production of CMCase by B. lincheniformis LBH-52 were investigated using the response surface method (RSM). The analysis of variance (ANOVA) of results from central composite design (CCD) indicated that a highly significant factor ("probe>F" less than 0.0001) for cell growth was rice hulls, whereas those for production of CMCase were rice hulls and initial pH of the medium. The optimal conditions of rice hulls, ammonium nitrate, initial pH, and temperature for cell growth extracted by Design Expert Software were 48.7 g/l, 1.8 g/l, 6.6, and 35.7$^{\circ}C$, respectively, whereas those for the production of CMCase were 43.2 g/l, 1.1 g/l, 6.8, and 35.7$^{\circ}C$. The maximal production of CMCase by B. lincheniformis LBH-52 from rice hulls under optimized conditions was 79.6 U/ml in a 7 l bioreactor. In this study, rice hulls and ammonium nitrate were developed to be substrates for the production of CMCase by a newly isolated marine microorganism, and the time for production of CMCase was reduced to 3 days using a bacterial strain with submerged fermentation.

• Korean Journal of Microbiology
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• v.30 no.5
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• pp.403-409
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• 1992

For the development of a glucanolytic yeast strain. the seceretion of endo-1.4-$\beta$-D-glucanase (CMCase) of Bacillus subtilis was performed in yeast using glucoamylase gene (STA1) of Saccharomyces diastaticus. A 1.7 kb-DNA fragment of STA1 gene containing authentic promoter, signal sequence, threonine serine-rich (TS) region and N-terminal region (98 amino acids) of mature glucoamylase was ligated to YEp 24. E. coli-yeast shuttle vector. And then. CMCase structural gene of B. subtilis was fused in frame with the 1.7 kb-DNA fragment of STA1 gene, resulting in recombinant plasmid pYES('24. Yeast transformant harboring pYESC24 had no CMCase activity. So. we deleted TS region and N-terminal region of mature glucoamylase existing between signal sequence and CMCase structural gene in pYESC24. consequently constructed recombinant plasmid pYESC11. The yeast transformed with the newly constructed recombinant plasmid pYESC11 efficiently secreted CMCase to extracellular medium. After 4 days culture. total CMCase activity of this transformant was 44.7 units/ml and over 93% of total CMCase activity was detected in culture supernatant.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.518-521
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• 1999

Bacillus sp. 79-23 spores were irradiated with $^{60}Co$ gamma-rays at doses ranging from 0.5 to 5 kGy. Following gamma-irradiation, seven mutant strains were isolated by scoring the halo sizes formed around the colonies grown on LB agar plates containing 4% carboxymethylcellulose (CMC) and trypan blue. The mutant strains showed a 1.5 to 2-fold increase in carboxymethylcellulase (CMCase) activity over the parent strain. Wheat bran acted as an effective inducer for CMCase production in the parent and mutant strains. Mutant strains 68 and 70 were identified as exhibiting higher CMCase activities than those of other mutants in LB media both with and without 3% wheat bran. In addition, these strains seem to produce substantially lower amounts of capsular materials, whereas the parent strain produced large amounts of them in both liquid and solid LB media. In flask cultures, the CMCase production by mutants 68 and 70 reached maximum levels of 17.5 unit/ml and 15.7 unit/ml, respectively, in an LB medium containing 3% wheat bran.

• Proceedings of the Korean Society of Life Science Conference
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• 2008.10a
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• pp.68.1-68.1
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• 2008