• Title/Summary/Keyword: Cellulomonas fimi

Search Result 12, Processing Time 0.023 seconds

Characteristics of Cellulomonas fimi $\beta$-glucosidase expressed in Escherichia coli (대장균에서 발현되는 Cellulomonas fimi $\beta$-glucosidase의 효소학적 특징)

  • Kim, Ha-Kun
    • The Journal of Natural Sciences
    • /
    • v.8 no.2
    • /
    • pp.57-61
    • /
    • 1996
  • The $\beta$-glucosidase enzyme was purified from E. coli carrying Cellulomonas fimi $\beta$-glucosidase gene. SDS-PAGE and analytical gel filtration revealed that molecular weight of this enzyme was 56,000 dalton and consisted of a single polypeptide.Inhibition caused by heavy metals and activation by dithiothreitol suggest the existence of essential thiol group in the enzyme. The enzyme was not active on maltose (glucose $\alpha$-1,4-glucose) which has a $\alpha$-linkage, whereas it was active on lactose (glucose $\beta$-1,4-glucose), PNPG (p-nitrophenyl $\beta$-D-glucopyranoside) and PNPC (p-nitrophenyl $\beta$-D-cellobioside), although its reaction rates were different.

  • PDF

Intergeneric protoplast fusion between Bacillus pumilus and Cellulomonas fimi

  • Kim, D.M.;Lee, K.H.
    • Proceedings of the Korean Society for Applied Microbiology Conference
    • /
    • 1986.12a
    • /
    • pp.528.1-528
    • /
    • 1986
  • Cellulose utilising hybrids between Cellulomonas fimi and Bacillus pumilus were isolated after PEG mediated protoplast fusion. 33% (w/v) PEG #6, 000 and 50mM $Ca^{++}$were optimum concentration. The intergeneric fusion frequency was 3.2$\times$10$^{-7}$ xtracellular CMCase and $\beta$-glucosidase activities were detected from one hydrid unlike only CMCase was detected from Cellulomonas fimi.

  • PDF

Isolation and Identification of Cellulomonas fimi, Characteristics of its Cellulase and Conversion of the Sawdust into Ethanol (Cellulomonas fimi의 분리(分離) 및 동정(同定), cellulase 특성(特性)과 톱밥의 Ethanol 전환(轉換))

  • Lee, Chan-Yong;Lee, Ke-Ho
    • Applied Biological Chemistry
    • /
    • v.28 no.2
    • /
    • pp.98-105
    • /
    • 1985
  • In the sheep and cattle's rumen, facultative anaerobic cellulolytic bacteria were isolated by using Hungate's roll tube technique. In the 21 isolated species, one was screened by its strong cellulolytic activity and identified as Cellulomonas fimi C-14 by investigate morphological, cultural, physiological characteristics and electron microgram. Optimum conditions of the cell growth and enzyme production were pH 6.5 an $30^{\circ}C$, Thiamine and biotin support a good growth of C. fimi C-14. In the enzyme activities, Crystalline cellulose hydrolyzing activity, CMCase activity and ${\beta}-glucosidase$ activity were 20.6, 226.6 and 0.56$(unit{\times}10^3/ml)$ at pH 6.0, $40^{\circ}C$. By addition of fungal cellulase, enzyme activity was increased. Simultaneous Saccharification Fermentation is better than two step fermentation in ethanol yield with Saccharomyces cerevisiae DY2.

  • PDF

Factors affecting the protoplast formation and regeneration of Bacillus pumilus and Cellulomonas fimi for intergeneric protoplast fusion

  • Kim, D.M.;Lee, K.H.
    • Proceedings of the Korean Society for Applied Microbiology Conference
    • /
    • 1986.12a
    • /
    • pp.527.3-527
    • /
    • 1986
  • Several factors predicted to affect the protoplast formation and regeneration were investigated. The optimum lysozyme, casamino acid and PVP concentration were 0.5 (mg/$m\ell$), 0.1 (%) and 1.5(%). In B. pumilus, Penicillin-G treatment concentration was 0.3 (U/$m\ell$) and optimum treatment period was transit log. phase. And in the case of Celm. fimi, 0.3 (U/$m\ell$) and initial log. phase. Osmotic stabilizer and di-cation for OSM medium of B.pumilus and Gelm .fimi were 25mM CaCl2, 0.5M sodium sucinate and 50mM MgCl$_2$, 100mM CaCl$_2$, 0.4M sodium succinate. The regeneration frequency of B.pumilus and Celm. fimi were 14.6(%) and 6.9(%).

  • PDF

Cloning and DNA Sequence of Carboxymethylcellulase (CMCase) Gene from Cellulomonas sp. YE-5

  • Her, Song;Kim, Dong-Seob;Choi, Sun-Jin
    • Journal of Microbiology and Biotechnology
    • /
    • v.3 no.2
    • /
    • pp.86-90
    • /
    • 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.

  • PDF

Studies on the Utilization of Agricultural Wastes (Part 2) Isolation and Identification of Cellulose Utilizing Bacteria. (농산폐자원의 이용에 관한 연구(제이보) 섬유소자화세균의 분리 및 동정)

  • Bae, Moo;Kim, Byung-Hong
    • Microbiology and Biotechnology Letters
    • /
    • v.2 no.1
    • /
    • pp.1-7
    • /
    • 1974
  • For the purpose of producing cellulosic single-cell protein from the agricultural wastes, 172 strains of cellulose-assimilating bacteria were isolated from 102 samples of rotten woods, compost soils, soils and so on by the enrichment culture technique. The isolates were examined for their ability to utilize cellulose as carbon source, and then six strains were screened by their strong cellulose assimilating ability and identified as follows: 1. Among six strains of bacteria screened, five strains were identified as species belonged to the genus Cellulomonas and the remainder to the genus of Sporocytophaga. 2. The isolated Sporocytophaga species was identified as S. ellipsosporn because it has a ellipsoidal microcyst. 3. The isolated Cellulomonas species were identical to a strain of C. fimi, C. aurcgena, C. gelida, respectively and two strains to C. flavigena. 4. The isolated C. aurogena was proved to be a new variety becauuse it has different characteristics of assimilating pentoses such as arabinose and xylose from the strain discribed in Bergey's Manual.

  • PDF

Taxonomic Studies on the Cellulolytic Bacterium Cellulomonas uda CS 1-1 (섬유소 분해균 Cellulomonas uda CS 1-1의 분류학적 연구)

  • Kim, Mi-Seog;Yoon, Min-Ho;Choi, Woo-Young
    • Korean Journal of Agricultural Science
    • /
    • v.34 no.2
    • /
    • pp.99-109
    • /
    • 2007
  • Cellulomanas sp. CS 1-1 was studied for its morphological, physiological and biochemical characteristics, together with DNA homology and fatty acid pattern to elucidate its taxonomical position in the species level. Colony morphology of CS1-1 exhibited circular form, opaque, convex, entire edge and pale yellow. Cells were of rod with the size of $0.3{\sim}0.5{\times}0.8{\sim}1.2{\mu}m$, while coryneforms were formed at the early stage of culture. D-ribose, raffinose, rhamnose, acetate, propionate, L-lactate, D-gluconate, aspartate and proline were not utilized as a sole source of carbon, whereas saccharose, arabinose, and amlyose were utilized. Biochemical characteristics of CS1-1 were Gram positive, catalase positive, oxidase negative, nonmotile, facultative anaerobic, mesophilic and G+C content of 74.7 mol %. The major fatty acid and menaquinone were 12-methyltetradecanoic acid(anteiso-$C_{15:1}$) and MK-$9(H_4)$, respectively. These results were correspondent with the characteristics reported for member of the genus Cellulomonas. The strain CS 1-1 exhibited a high level of DNA homology as 70% with C. uda ATCC491, compared to those of 54~59% with C. fimi ATCC 15724, 46~48% with C. biazotea, C. gelida and C. bibula. Finally, strain CS1-1 could be classified as a novel species belongs to C. uda.

  • PDF

Studies on Microbial Utilization of Agricultural Wastes (Part IX) Analsis of Cellulosic Single Cell Protein and Their Amino Acid Patterns (농산방자원의 이용에 관한 연구 (제9유) 섬유소단세포단백질의 아미노산 조성 및 그의 영향학적 가치)

  • Kim, Byung-Hong;Bae, Moo
    • Microbiology and Biotechnology Letters
    • /
    • v.5 no.4
    • /
    • pp.167-169
    • /
    • 1977
  • Cellulosic single cell proteins produced from rice straw by three strains of cellulolytic bacteria were analysed for their composition and for their amino acid patterns. It was showed that they contains 52.2∼70.3% of crude Protein, 11.6∼13.9% of crude fat and 11.5∼18.1% of ash. The sulphur containing amino acids and threonine were analysed to be the limiting amino acid in the cellulosic SCP. The protein scores were calculated as 80.0 for Cellulomonas flavigena KIST 321, 78.00 for Cellulomonas fimi and 61.8 for Cellulomonas aurogena KIST 11. The nutritional value of the cellulosic SCP is discussed from the results.

  • PDF

Overproduction and Secretion of $\beta$-Glucosidase in Bacillus subtilis

  • Kim, Jeong-Hyun;Lee, Baek-Rak;Moo, young-Pack
    • Journal of Microbiology and Biotechnology
    • /
    • v.8 no.2
    • /
    • pp.141-145
    • /
    • 1998
  • Overproduction of intracellular ${\beta}$-glucosidase was attempted by modifying the promoter region of a ${\beta}$-glucosidase gene cloned from Cellulomonas fimi and expressing it in Bacillus subtilis DB 104. A strong engineered promoter, BJ27UΔ88, was fused to the ${\beta}$-glucosidase gene after removing its native promoter. An effective Shine-Dalgamo sequence (genel0 of phage T7) was inserted between the promoter and the ${\beta}$-glucosidase structural gene. The modified gene was overexpressed in B. subtilis and produced 1121.5 units of ${\beta}$-glucosidase per mg protein which is about $12\%$ of total intracellular protein. Secretion of overproduced intracellular ${\beta}$-glucosidase was attempted by using the signal sequence of the Bacillus endoglucanase gene as well as an in-frame hybrid protein of endoglucanase. The hybrid protein was normally secreted into the culture medium and still retained ${\beta}$-glucosidase activity.

  • PDF

Screening of Methotrexate-Resistant Strains with High Thymidylate Synthase Activity (티미딜산 생성효소 활성이 높은 메토트렉세이트-내성 균주의 검색)

  • Kwak, In-Young;Lee, Jong-Soo
    • YAKHAK HOEJI
    • /
    • v.36 no.4
    • /
    • pp.345-349
    • /
    • 1992
  • Thymidylate synthase activity from extracts of various methotrexate-resistant strains was measured by spectrophotometric assay. Methotrexate-resistant strains of Lactobacillus, Pseudomonas sp., Micrococcus sp. HS-1, Klebsillela pneumonae, Cellulomonas fimi and Serratia marcescens elevated thymidylate synthase levels, especially, Pseudomonas sp. KL-9 resistant to $10^{-9}M$ methotrexate have a 156-fold increase in thymidylate synthase, which suggests that Pseudomonas sp. is a convenient source of thymidylate synthase. Several methotrexate strains of yeast were tested, however, their enzyme activity was generally lower than that of bacteria tested.

  • PDF