• Title/Summary/Keyword: cellulase gene

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Physical and catalytic properties of CMCase encoded by Bacillus subtilis gene in B. megaterium

  • Kim, Hoon;Kim, Ha-Geun;Park, Moo-Young
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.524.3-524
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    • 1986
  • Carboxymethyl cellulase (CMCase) produced by cloned B. megaterium was found to contain 5.2% carbohydrate but no metal ion. The enzyme was isoelectric at pH 7.23 and was high is basic amino acids. The N-terminal of the enzyme was glutamic acid. The cellulolytic activity of this enzyme was extended to the small molecular substrates such as from cellotriose to cellopentaose. In additon, the enzyme showed transglycoslation activity. The pK values of the enzyme we estimated to be 4.4 and 6.7, andthat of the enzyme-substrate complex were 4.2 and 7.2, respectively. The enzyme was not affected by the treatment with iodoacetic acid, but the modification of enzyme with carbodiimide and diethyl pyrocarbonate resulted in a marked loss of the enzyme activity. These results suggest that the active site of enzyme essentially contains carboxylic and imidazole group of amino acid residues.

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Biocontrol of Fusarium Crown and Root Rot and Promotion of Growth of Tomato by Paenibacillus Strains Isolated from Soil

  • Xu, Sheng Jun;Kim, Byung Sup
    • Mycobiology
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    • v.42 no.2
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    • pp.158-166
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    • 2014
  • In this study, bacterial strains were isolated from soils from 30 locations of Samcheok, Gangwon province. Of the isolated strains, seven showed potential plant growth promoting and antagonistic activities. Based on cultural and morphological characterization, and 16S rRNA gene sequencing, these strains were identified as Paenibacillus species. All seven strains produced ammonia, cellulase, hydrocyanic acid, indole-3-acetic acid, protease, phosphatase, and siderophores. They also inhibited the mycelial growth of Fusarium oxysporum f. sp. radicis-lycopersici in vitro. The seven Paenibacillus strains enhanced a range of growth parameters in tomato plants under greenhouse conditions, in comparison with non-inoculated control plants. Notably, treatment of tomato plants with one identified strain, P. polymyxa SC09-21, resulted in 80.0% suppression of fusarium crown and root rot under greenhouse conditions. The plant growth promoting and antifungal activity of P. polymyxa SC09-21 identified in this study highlight its potential suitability as a bioinoculant.

On a highly proteolytic mutant strain of Aspergillus flavus (Aspergillus flavus의 강력 protease생성 돌연변이의 유발)

  • 이영녹;박용근;고상균
    • Korean Journal of Microbiology
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    • v.18 no.2
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    • pp.51-58
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    • 1980
  • Mutational experiments were performed to improved to improve the protease productivity of Aspergillus flavus KU 153, which is selected among the wild strains. A UV-induced mutant strain having high protease productivity was obtained by the use of the clear zone method as a simple criterion for a primary screening test. Neutral and alkaline protease activities of hte mutant strain were higher than 1.8 times, comopared with those of the parental strain, respectively, while in the case of acid protease, it was 2.7 times. The mutant strain selected was more powerful in the production of cellulase and amylase, as well s protease in wheat bran, compared with those of the parental strain. protease production of the parental strain has reached maximum level at 3 days culture, while alkaline nad neutral protease production of the mutantstrain has reached at 2 days culture. On the other hand, the mutant strain formed the spore slowly, compared with the parental strain. Column chromatography of the neutral protease on DEAE-Sephadex A-50 showed that the mutant strain was not induced the formation of another neutral protease isozyme, but induced the variation in the function of regulatory gene.

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Complete genome sequence of Paenibacillus konkukensis sp. nov. SK3146 as a potential probiotic strain

  • Jung, Hae-In;Park, Sungkwon;Niu, Kai-Min;Lee, Sang-Won;Kothari, Damini;Yi, Kwon Jung;Kim, Soo-Ki
    • Journal of Animal Science and Technology
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    • v.63 no.3
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    • pp.666-670
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    • 2021
  • Paenibacillus konkukensis sp. nov., SK3146 is a novel strain isolated from a pig feed. Here, we present complete genome sequence of SK3146. The genome consists of a single circular genome measuring 7,968,964 bp in size with an average guanine + cytosine (G+C) content of 53.4%. Genomic annotation revealed that the strain encodes 151 proteins related to hydrolases (EC3), which was higher than those in Bacillus subtilis and Escherichia coli. Diverse kinds of hydrolases including galactosidase, glucosidase, cellulase, lipase, xylanase, and protease were found in the genome of SK3146, coupled with one bacteriocin encoding gene. The complete genome sequence of P. konkukensis SK3146 indicates the immense probiotic potential of the strain with nutrient digestibility and antimicrobial activity functions.

Characterization of Actinomyces Isolated from Freshwater Sponges in Lake Baikal (바이칼의 담수 스폰지에서 분리한 방선균의 특성 연구)

  • Jung, You-Jung;Joung, Yo-Chan;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.47 no.2
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    • pp.130-136
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    • 2011
  • Five strains of Actinomyces were isolated from freshwater sponges, Baikalospongia and Lubomirskia, in Lake Baikal. By 16S rRNA sequencing, isolates were identified as Streptomyces griseoplanus, S. halstedii, S. violascens, S. flavovirens, and S. microflavus. Isolates had different characteristics of growth temperature, carbon utilization, enzyme activity, and cellular fatty acid composition. Optimum growth conditions of isolates were $30-37^{\circ}C$, pH 8-9, and 0-1.5% salt concentrations. Major fatty acid compositions were anteiso-$C_{15:0}$, iso-$C_{15:0}$, and iso-$C_{16:0}$. Strain ATS-BA-19 had DNase and chitinase activities and strain ATS-BA-16 had cellulase and protease activities. Colonies of strain ATS-BA-15 and ATS-BA-19 made inhibition zone of Pseudomonas aeruginosa.

Virulence Reduction and Differing Regulation of Virulence Genes in rpf Mutants of Xanthomonas oryzae pv. oryzae

  • Jeong, Kyu-Sik;Lee, Seung-Eun;Han, Jong-Woo;Yang, Seung-Up;Lee, Byoung-Moo;Noh, Tae-Hwan;Cha, Jae-Soon
    • The Plant Pathology Journal
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    • v.24 no.2
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    • pp.143-151
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    • 2008
  • To define the functions of the rpf genes in Xanthomonas oryzae pv. oryzae (Xoo), which regulates pathogenicity factors in Xanthomonas campestris pv. campestris (Xcc), marker-exchange mutants of each rpf gene were generated. When the mutants were inoculated on a susceptible cultivar, the lesion lengths caused by the rpfB, rpfC, rpfF, and rpfG mutants were significantly smaller than those caused by the wild type, whereas those caused by the rpfA, rpfD, and rpfI mutants were not. Several virulence determinants, including extracellular polysaccharide (EPS) production, xylanase production, and motility, were significantly decreased in the four mutants. However, the cellulase activity in the mutants was unchanged. Complementation of the rpfB and rpfC mutations restored the virulence and the expression of the virulence determinants. Expression analysis of 14 virulence genes revealed that the expression of genes related to EPS production (gumG and gumM), LPS (xanA, xanB, wxoD, and wxoC), phytase (phyA), xylanase (xynB), lipase (lipA), and motility (pitA) were reduced significantly in the mutants rpfB, rpfC, rpfF, and rpfG. In contrast, the expression of genes related to cellulase (eglxob, clsA), cellobiosidase (cbsA), and iron metabolism (fur) was unchanged. The results of this study clearly show that rpfB, rpfC, rpfF, and rpfG are important for the virulence of Xoo KACC10859, and that virulence genes are regulated differently by the Rpfs.

Development of Cell Lines for Application of Recombinant DNA Techniques in Crops (작물의 유전자 재조합을 위한 세포주의 개발 연구)

  • Chae, Young-Am;Choi, Kyu-Whan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.2
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    • pp.195-200
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    • 1985
  • This experiment was carried out to know the processes of protoplast isolation, culture and plant regeneration in aims of introducing foreign genes into plant cells through plant gene vector, and cellular selection for plant improvement. The main results indicated that 2% cellulase plus 0.5% macerozyme is proper for isolation of protoplasts from leaf mesophyll cells of N. plumbaginifolia, plating efficiency was higher in 1.4-2.0 x 10$^4$ cells/ml, complete cell wall was regenerated after 2 days culture, cell division and cell mass were observed after 4 days and 2 weeks, respectively, colony was developed after 3 weeks culture, addition of 1-2mg/l BA promoted shoot differentiation while root differentiation did not required hormone and seeds were harvested from more than 100 cell lines for further investigation and study.

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Functional Analysis of a Gene Encoding Endoglucanase that Belongs to Glycosyl Hydrolase Family 12 from the Brown-Rot Basidiomycete Fomitopsis palustris

  • Song, Byeong-Cheol;Kim, Ki-Yeon;Yoon, Jeong-Jun;Sim, Se-Hoon;Lee, Kang-Seok;Kim, Yeong-Suk;Kim, Young-Kyoon;Cha, Chang-Jun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.404-409
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    • 2008
  • The brown-rot basidiomycete Fomitopsis palustris is known to degrade crystalline cellulose (Avicel) and produce three major cellulases, exoglucanases, endoglucanases, and ${\beta}$-glucosidases. A gene encoding endoglucanase, designated as cel12, was cloned from total RNA prepared from F. palustris grown at the expense of Avicel. The gene encoding Cel12 has an open reading frame of 732 bp, encoding a putative protein of 244 amino acid residues with a putative signal peptide residing at the first 18 amino acid residues of the N-terminus of the protein. Sequence analysis of Cel12 identified three consensus regions, which are highly conserved among fungal cellulases belonging to GH family 12. However, a cellulose-binding domain was not found in Cel12, like other GH family 12 fungal cellulases. Northern blot analysis showed a dramatic increase of cel12 mRNA levels in F. palustris cells cultivated on Avicel from the early to late stages of growth and the maintenance of a high level of expression in the late stage, suggesting that Cel12 takes a significant part in endoglucanase activity throughout the growth of F. palustris. Adventitious expression of cel12 in the yeast Pichia pastoris successfully produced the recombinant protein that exhibited endoglucanase activity with carboxymethyl cellulose, but not with crystalline cellulose, suggesting that the enzyme is not a processive endoglucanase unlike two other endoglucanases previously identified in F. palustris.

Gene Cloning of Cellulose Degradation Enzyme of Bacillus subtilis LYH201 Strain (Bacillus subtilis LYH201균주의 섬유소 분해효소의 유전자 Cloning 및 특성분석)

  • Lee, Young-Han;Park, Sang-Ryeol
    • Korean Journal of Soil Science and Fertilizer
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    • v.34 no.5
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    • pp.333-341
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    • 2001
  • The Compost-decomposing-bacteria was isolated from livestock compost containing sawdust. The isolated bacteria was identified as Bacillus subtilis LYH201 by the method of the composition of the fatty acid with MIDI system and Bergey's manual. Cloning of CMCase encoding gene was accompanied by shotgun method. The pLK100 have yellow activity ring on CMC medium, that was carried 2.2 kb insert DNA in pBluescript II $SK^+$ vector, named BglC gene. The BglC was very similar to Pectobacterium carotovorum Gun_CLOAB(P15704) with score of 57% identity and 71% homology over 508 aa. The BglC was measured molecular weight 56 kDa by CMC-SDS-PAGE. Optimum cellulase activity Bacillus subtilis LYH201 was temperature $50^{\circ}C$ and pH 7.

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