• Title/Summary/Keyword: 3-Glucanase

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Effect of $Glucanex^{(R)}$ 200G on Yeast Cell Wall (효모의 세포벽에 대한 $Glucanex^{(R)}$ 200G의 영향)

  • 김광석;장정은;윤현식
    • KSBB Journal
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    • v.19 no.4
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    • pp.288-290
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    • 2004
  • The cell wall of fifteen yeast strains were treated with Glucanex$^{(R)}$ 200G that contained mainly ${\beta}$1,3-glucanase and some ${\beta}$1,6-glucanase. In our previous study it was found that the yeasts that are more resistant to Glucanex$^{(R)}$ 200G treatment contained more ${\beta}$-glucan than the yeasts that are less resistant to Glucanex$^{(R)}$200G treatment. By measuring the resistance of cell wall to Glucanex$^{(R)}$ 200G, the relative content of ${\beta}$-glucan in yeast cell wall could be estimated. The resistance of cell wall to Glucanex$^{(R)}$ 200G was measured by counting viable cell number after reaction with and without Glucanex$^{(R)}$200G. The resistance of fifteen yeast strains to Glucanex$^{(R)}$ 200G were presented.ere presented.

Antifungical Activity of Autochthonous Bacillus subtilis Isolated from Prosopis juliflora against Phytopathogenic Fungi

  • Abdelmoteleb, Ali;Troncoso-Rojas, Rosalba;Gonzalez-Soto, Tania;Gonzalez-Mendoza, Daniel
    • Mycobiology
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    • v.45 no.4
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    • pp.385-391
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    • 2017
  • The ability of Bacillus subtilis, strain ALICA to produce three mycolytic enzymes (chitinase, ${\beta}$-1,3-glucanase, and protease), was carried out by the chemical standard methods. Bacillus subtilis ALICA was screened based on their antifungal activity in dual plate assay and cell-free culture filtrate (25%) against five different phytopathogenic fungi Alternaria alternata, Macrophomina sp., Colletotrichum gloeosporioides, Botrytis cinerea, and Sclerotium rolfesii. The B. subtilis ALICA detected positive for chitinase, ${\beta}$-1,3-glucanase and protease enzymes. Fungal growth inhibition by both strain ALICA and its cell-free culture filtrate ranged from 51.36% to 86.3% and 38.43% to 68.6%, respectively. Moreover, hyphal morphological changes like damage, broken, swelling, distortions abnormal morphology were observed. Genes expression of protease, ${\beta}$-1,3-glucanase, and lipopeptides (subtilosin and subtilisin) were confirmed their presence in the supernatant of strain ALICA. Our findings indicated that strain ALICA provided a broad spectrum of antifungal activities against various phytopathogenic fungi and may be a potential effective alternative to chemical fungicides.

Simultaneous Expression of Pseudomonas sp. Endo-1,4$\beta$-Glucanase and $\beta$-1,4=Glucisidase Gene in Escherichia coli and Saccharomyces cerevisiae (Pseudomonas sp. Endo-1,4-$\beta$-Glucanase와 $\beta$-1,4-Glucosidase 유전자의 대장균 및 효모에서의 동시 발현)

  • Kim, Yang-Woo;Chun, Sung-Sik;Chung, Young-Chul;Sung, Nack-Kie
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.652-658
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    • 1995
  • We attempted simultaneous expression of genes coding for endoglucanase and $\beta $-glucosidase from Pseudomonas sp. by using a synthetic two-cistron svstem in Escherichia coli and Saccharomyces cerevisiae. Two-cistron system, 5'--tac promoter-endoglucanase gene--$\beta $-glucosidase gene-- 3', 5'-tac promoter--$\beta $-glucosidase gene--endoglucanase gene--3' and 5'-tac promoter--endoglucanase gene--SD sequence--$\beta $-glucosidase gene--3, were constructed, and expressed in E. coli and S. cerevisiae. The E. coli and S. cerevisiae contained two-cistron system produced simultaneously endoglucanase and $\beta $-glucosidase. The recombinant genes contained the bacterial signal peptide sequence produced low level of endoglucanase and $\beta $-glucosidase in S. cerevisiae transformants: Approximately above 44% of two enzymes was localized in the intracellular fraction. The production of endoglucanase and $\beta $-glucosidase in veast was not repressed in the presence of glucose or cellobiose. The veast strain contained recombinant DNA with two genes hydrolyzed carboxvmethyl cellulose, and these endoglucanase and $\beta $-glucosidase degraded CMC synergistically to glucose, cellobiose and oligosaccharide. This result suggests the possibility of the direct bioconversion of cellulose to ethanol by the recombinant yeast.

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Purification and Characterization of Degradative Enzyme of Dental Plaque from Streptomyces sp. Y9343 (Streptomyces sp. Y9343이 生産하는 齒面細菌膜 分解酵素의 精製와 特性)

  • Kim, Seong-Joo;Han, Hong-Keun;Yoon, Jeong-Weon
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.9-18
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    • 1996
  • Streptococcus mutans has been implicated as primary causative agents of dental caries by insoluble glucan (IG) in human and experimental animals. An attempt was made to search for the ${\alpha}$-1,3 glucanase that degrades IG produced by S. mutans. ${\alpha}$-1,3 glucanase was detected in the culture supernatant of microorganisms, which are isolated from soils on agar medium containing IG as a sole carbon source. This Streptomyces sp. hydrolysed IG produced by immobilized S. mutans and was named as Y9373. This enzyme required ${\alpha}$-1,3 glucan (IG) as an inducer. The optimum conditions for enzyme production were studied. The enzyme was purified by 30~70% $(NH_4)_2SO_4$ precipitation, anion exchange chroma tography on DEAE-cellulose and gel filtration on Sepadex G-75. The purified enzyme has a specific activity of 7840.0 U/mg protein giving 32.1-fold purification and final yield of 0.53%. The molecular weight was estimated to be about 22.5 kDa by SDS-PAGE. The optimum pH and temperature for enzyme reaction were 6.5 and 37$^{\circ}C$, respectively and the enzyme was relatively stable at the temperature below 60$^{\circ}C$. The activity of purified enzyme was enhanced by adding $Co^{2+},\;Mn^{2+}\;and\;Mg^{2+}$ into the medium, whereas inhibited by adding $Hg^{2+},\;Zn^{2+}$ and SDS. The $K_m\;and\;V_{max}$ value of ${\alpha}$-1,3 glucanase for IG were estimated to be 2.50 mM and 0.0431 mM/min, respectively. The thin layer chromatographic analysis of hydrolysates from IG with ${\alpha}$-1,3 glucanase showed that glucose was the main product of reaction. This enzyme activity was about 14 times higher than marketing dextranase as preventive agent against artificial dental caries by S. mutans in TH medium including 5% sucrose after 30 minutes.

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Molecular cloning of the Arabidopsis gene rat3 that is involving in the Agobacterium-mediated planttransformation (Agrobacterium에 의한 식물형질전환에 관여하는 Arabidopsis RAT3 유전자의 분리와 분석)

  • 남재성;양보경;김도훈;정순재;이영병
    • Journal of Life Science
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    • v.11 no.5
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    • pp.423-431
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    • 2001
  • Genomic and cDNA clones containing the RAT3 gene involving in Agobacterium-mediated plant transformation were identified using plant DNA flanking the righ border of a T-DNA rescued from the rat3 mutant as hy-bridization probe. Two highly homologous cDNA clones were identified; one (RAT3-1) weakly hybridized with the probe whereas another (RAT3-2) strongly hybridized with the probe. Both Rat3-1 and Rat3-2 proteins contain a putative signal peptide for secretion. The deduced molecular weights of encoded proteins are 15 kDa. The results of genomic DNA blot analysis and DNA sequencing indicated that RAT3-1 and RAT3-2 exist as single copy genes and they were arranged side by side with just 600 bp distance between them. RAT3-1 was disrupted by the integration of T-DNA into the 3 untranslated region in rat3 mutant. A BLAST search showed that both RAT3-1 and RAT3-2 proteins have homology with only the C-terminal region of $\beta$-1,3-glucanase homologues from Triticum aestivum and Arabidopsis thaliana. Thses $\beta$-1,3-glucanase homologues contain an unusually long C-terminal region with no sig-nificant homology to other $\beta$-1,3-glucanase.

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Isolation and Characterization of Endo-$\beta$-1,4-glucanase from the Midgut of the Earthworm, Eisenia andrei (지렁이 중장에서 발현되는 Endo-$\beta$-1,4-glucanase의 동정 및 특성에 관한 연구)

  • Lee Myung Sik;Cho Sung Jin;Tak Eun Sik;Hur So Young;Lee Jong Ae;Park Bum Joon;Cho Hyun Ju;Shin Chuog;Park Soon Cheol
    • The Korean Journal of Soil Zoology
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    • v.8 no.1_2
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    • pp.7-12
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    • 2003
  • Endogeneous endoglucanase (EC 3.2.1.4) cDNA was cloned from a representative species (Eisenia anderi) of the earthworm family Lumbricidae. Endoglucanase from the midgut of the earthworm is composed of 456 amino acids and belongs to glycosyl hydrolase family 9 (GHF9), sharing high homologies (50-51 %) with those of selected termite and crayfish. This endoglucanase consists of three consensus catalytic domains found in most microbial cellulases. A phylogenetic tree was constructed using the amino acid squence data matched through the BLASTX program and showed that GHF9 families could be divided into four groups of arthropoda, bacteria, plant and annelida.

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Studies on the Enzymatical Properties of Streptomyces sp. S-45 Isolated from Soil (토양(土壤)에서 분리(分離)한 Streptomyces sp. s-45의 효소학적(酵素學的) 성질(性質)에 관한 연구(硏究))

  • Kim, Yeong-Yil;Kim, Yong-Woong;Kim, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.2
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    • pp.129-134
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    • 1988
  • Enzymatical properties of Streptomyces sp. S-45 producing chitinase and ${\beta}$-1.3-glucanase isolated from soil were investigated. Chitinase activity was 3.01(U/ml) and ${\beta}$-1.3-glucanase activity was 2.49(U/ml). The optimum medium for mycolytic enzyme production of strain was composed of 0.7% colloidal chitin, 0.3% glucose, 0.5% asparagine, 0.2% peptone, 0.01% NaCl, 0.01% $K_2HPO_4$ and 0.01% $MgSO_4{\cdot}7H_2O$ in intial pH 7.0. The optimal condition for mycolytic enzyme activities were: pH 6.5-7.0, $45-50^{\circ}C$. Enzyme activities were activated by metal ion as $10^{-2}M\;Co^{{+}{+}}$, $Cu^{{+}{+}}$, $Mn^{{+}{+}}$, $Al^{{+}{+}{+}}$ and $10^{-3}M\;Sn^{{+}{+}}$ but $Ag^{{+}{+}}$, $Hg^{{+}{+}}$ inhibited.

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Roles of Carbohydrate-Binding Module (CBM) of an Endo-β-1,4-Glucanase (Cel5L) from Bacillus sp. KD1014 in Thermostability and Small-Substrate Hydrolyzing Activity

  • Lee, Jae Pil;Shin, Eun-Sun;Cho, Min Yeol;Lee, Kyung-Dong;Kim, Hoon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2036-2045
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    • 2018
  • An endo-${\beta}$-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and $55^{\circ}C$, but had different half-lives of 4.0 and 22.8 min, respectively, at $70^{\circ}C$. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ${\beta}$-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

Pilot-Scale Production of Cellulase Using Trichoderma reesei Rut C-30 Fed-Batch Mode

  • Lee, Sang-Mok;Koo, Yoon-Mo
    • Journal of Microbiology and Biotechnology
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    • v.11 no.2
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    • pp.229-233
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    • 2001
  • Trichoderma reesei Rut C-30 produced high levels of ${\beta}$-glucosidase, endo-${\beta}$-glucosidase, endo-${\beta}$-1,4-glucanase, and exo-${\beta}$-1,4-glucanase. In pilot-scale production (50-1 fermentor), productivity and yield of CMCase (carborymethyl cellulose) and FPase (filter paper activity) were 273 U/ml and 35 U/ml, and 162 FPU/l.h and 437 FPU/g, respectively. The fed-batch techniques were used to improve enzyme activities with constant cell concentration. The acidity was an important parameter and controlled at pH 3.9 and 5.0 by automatic addition of ammonium hydroxide. Cellulase powder was prepared by ammonium sulfate precipitation and its CMCase and FPase activities were 3,631 U/g and 407 U/g, respectively.

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