• Journal of Microbiology and Biotechnology
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• v.3 no.2
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• pp.86-90
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• 1993

CMCase positive clones were screened from Cellulomonas sp. YE-5 and named pCE1, pCE2 and pCE3. Among the positive clones pCE1 was used for this study, because it has the smallest insert and the highest CMCase activity among the 3 clones, and its nucleotide sequence was determined. The CMCase gene in pCE1 was composed of 1071 bp of nucleotides coding 357 amino acids. Computer analysis showed that the pCE1 has 65% sequence homology with the endoglucanase from Cellulomonas fimi.

• Microbiology and Biotechnology Letters
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• v.18 no.4
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• pp.376-382
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• 1990

An extracellular cellulase producing bacterium YE-5 was isolated from soil, and identified as a Cellulomonas sp. by its taxonomical characteristics. The maximal activities of avicelase (0.35 units/ml), CMCase (3.18 units/ml), FPase (0.315 units/ml) and $\beta$-glucosidase (0.882 units/ml) were obtained when this strain was cultured for 48 hrs at $30^{\circ}C$ in a medium containing 0.8% (w/v) Solka floc, 0.06010 (wlv) urea, 0.1% (w/v) $K_2HP0_4$, 0.1% (w/v) $MgS0_4.7H2_0$, 0.2% (w/v) bacto peptone, 0.2% (w/v) yeast extract and pH 6.5. The cellulase was purified by ammonium sulfate fractionation, DEAE-Sepharose column chromatography and Sephadex 6-100 column chromatography from culture filtrate of Cellulomonus sp. YE-5. The molecular weights of purified avieelase, CMCase I, and CMCase II were estimated to be about 95,000 ~ 105,000, 46,000 ~ 47,000 and 120,000 ~ 125,000, respectively.

• Microbiology and Biotechnology Letters
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• v.6 no.2
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• pp.51-57
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• 1978

The study was made of the cultural condition and physiological characteristics of the symbiotic pair of microorganisms, Cellulomonas flavigena and the second organism. It also contains the results of a taxonomical study of the second organism. The results obtained wers summarized as follows : 1) Cell yield of the mixed culture, Cellulomonas and the second organism, was higher than that of each pure culture in CM-Cellulose medium. 2) The taxonomical characteristics of the second organism revealed that it probably belonged to the genus Sporocytophaga because it had a gliding motility and microcyst. 3) Optimum pH of the mixed culture was found to be in the vicinity of 7.2, and optimum temperature of the cell growth in the mixed culture was observed to be in the vicinity of 30$^{\circ}C$. 4) It was found that the majority of the population during growth in the mixed culture consisted of Cellulomonas flavigena. 5) Cellulomonas flavigena required thiamine and biotin as growth factors but Sporocytophaga sp. had no requirement of vitamins. 6) Gulucose was not found in detectable amounts in the medium of Cellulomonas flavigena but it was traced in the mixture by thin layer chromatography. 7) Sixteen amino acids were analyzed from the cell protein of Cellulomonas flavigena by amino acid autoanalyzer. The amount of the leucine, valine and arginine was very high.

• Korean Journal of Agricultural Science
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• v.3 no.2
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• pp.225-234
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• 1976

To elucidate ${\beta}$-glucosidase formation on various carbon scurces by cellulolytic bact-eia, Cellulomonas sp. CS1-1, the strain was grown on Nutrient Yeast Broth, carboxymethyl cellulose, avicel and cellobiose using a Ouickfit FVIL fermentor operated in batch, and the growth characteristics on those substrates and ${\beta}$-glucosidase distribution of extra and intracellular enzyme components were studied. The results were: 1) ${\beta}$-glucosidase was always intracellular, and was formed under all growth conditions tested, ii) but levels of relative activities were higher when the culture was grown on cellobiose and on avicel, iii) the relative activities were always maximum during the growth phase of the organism irrespective of the substrate used.

• Korean Journal of Microbiology
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• v.23 no.4
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• pp.309-314
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• 1985

Autoxotrophic mutants of a cellulolytic baterium Cellulomonas sp. CS 1-1 were grown at $30^{\circ}C$ for 6hr using a complete medium containing 0.5M sucrose and for another 90 min after addition of 0.3 U/ml penicillin G, and were protoplasted by 0.2mg/ml lysozyme for 2hr. Prototrophic recombinants were obtained at the rates of $10^{-3}$ to $10^{-5}$by fusing the protoplasts in the presence of 40% polyethyleneglycol3350. Nystatin could be used to eliminate fungal contamination during the regeneration of the plotaplasts.

• Applied Biological Chemistry
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• v.38 no.6
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• pp.490-495
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• 1995

Among the cellulases by Cellulomonas sp. KL-6. CMCase and filter paperase, which were produced as the out enzymes of cell, had been much produced, but very small amounts of ${\beta}-glucosidase $, the enzyme of which is cell bound form, was produced by this organism. The optimal culture times for CMCase and filter paperase productions were 5 days, while that of ${\beta}-glucosidase$ was 4 days. When this strain was cultured under the optimal medium for enzyme production, CMCase, FPase and ${\beta}-glucosidase$ were $82\;units/m{\ell},\;80\;units/m{\ell}\;and\;1.2\;units/m{\ell}$, respectively. Thus these results were showed to increase enzyme productivities as about $60{\sim}70%$ than those produced in basal medium. $CaCO_3$ injected to the medium as the ratio of 0.1% was not only enhanced cellulase activities but also effective as acid neutralizing agent. The production effects of lignase and lactase by this bacterium in filter paper medium was not appeared.

• Journal of Microbiology and Biotechnology
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• v.14 no.6
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• pp.1196-1199
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• 2004

A bacterium, isolated from rabbit's waste and identified as Cellulomonas sp., had cellulase and thermostable $\alpha$-amylase activity when grown on wheat bran. Maximum activity of thermostable $\alpha$-amylase was obtained by adding $3\%$ soluble starch. However, soybean oil (1 ml $1^{-1}$) could increase the production of $\alpha$-amylase and cellulase in 'wheat bran. The $\alpha$-amylase was characterized by making a . demonstration of optimum activity at $90^{\circ}C$ and pH 6- 9, with soluble starch as a substrate. The effect of ions on the activity and the stability of this enzyme were investigated. This strain secreted carboxymethyl cellulase (CMCase), cellobiase ($\beta$­glucosidase), and filter paperase (Fpase) during growth on wheat bran. Carboxymethy1cellulase, cellobiase, and Fpase activities had pH optima of 6, 5.5, and 6, respectively. CMCase and cellobiase activities both had an optimum temperature of $50^{\circ}C$, whereas Fpase had an optimum temperature of $45^{\circ}C$.

• Korean Journal of Microbiology
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• v.22 no.3
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• pp.167-173
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• 1984

The cellabiase (${\beta}$-glucosidase) gene in a Cellulomonas sp. CS1-1 was cloned into E. coli HB101 using the vector plasmid pBR322, and the expression of the gene in E. coli studied. The chromosomal DNA of the cellulomonas was digested by seveal restriction enzymes, each of which has only one cleaving site in plasmid pBR322. The recombinant plasmid, pSB2, created with Sal I frament, was expressed for the cellobiase gene in E. coli. The recombiant plasmid was estimated to contain 6.4 Kb foreign DNA at the Sal I site of plasmid pBR322 and the inserted DNA was mapped by single and double digestion with several enzymes. E. coli HB101(pSB2) has slowly grown in a mineral liquid medium containing cellobiose as a sole carbon source. The cellobiase activity in the transformed E. coli was 132 units per liter, which is equivalent to one twenty fifth of that in doner strain Cellulomonas sp. CS1-1. The transforned cell with plasmid containing cellulase gene grow well in the LB mediuns. The synthesis of cellobiase in the strain, E. coli HB101 (pSB2), was inhibited by glucose and at high concentration of cellobiose, and induced by cellobiose at low concentration.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 1986.12a
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• pp.525.1-525
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• 1986

To clone ${\beta}$-glucosidase gene from Cellulomonas sp. a gene library was constructed using E. coli JM83 pUC9. Among 2,500 pseudotransformants obtained, 20 clones developed yellow color on the p-nitrophenyl- -D-glucopyranoside filter paper These 20 clones were classified into three groups based on the results of activity staining using nondenaturating polyacrylamide gel electrophoresis and restriction enzyme digestions. Among the three groups, only one group containing pCEl plasmid has specificity for cellobiose.

• Microbiology and Biotechnology Letters
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• v.20 no.2
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• pp.164-168
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• 1992

Enzymatic properties of avicelase, carboxymethyl cellulase (CMCase) and P-glucosidase produced by Cellulomonas sp. YE-5 were studied. Optimal temperature and pH of avicelase were 40t and 6.0, and those of CMCase and P-glucosidase were $45^{\circ}C$ and 6.5. Avicelase and CMCase were stable between pH 5.0 and 9.5, and &glucosidase was stable between pH 5.5 and 8.0. Avicelase and P-glucosidase were inactivated when incubated at $35^{\circ}C$ for 6 hrs, and CMCase was at $40^{\circ}C$ for 6 hrs. All cellulases were strongly inhibited by $Cu^{2+} \; and \; Zn^{2+}. K_m$ values of avicelase for avicel, CMCase I and CMCase II for CM-cellulose, and ($\beta$-glucosidase for p-nitrophenyl-$\beta$-D-glucoside (PNPG) were 4.76, 16.4, 16.4 $\mu g$/ml and 3.51 mM, respectively.