• Title/Summary/Keyword: extracellular chitinase

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Studies on the Classification and Identification of Actinomycetes Producing Chitinase isolated from Soil (토양(土壞)에서 분리(分離)한 Chitinase 생성(生成) 방선균(放線菌)의 분류동정(分類同定))

  • Kim, Yeong-Yil;Kim, Yong-Woong;Kim, Kwang-Sick
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.3
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    • pp.339-345
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    • 1988
  • Four actinomycetes which produces extracellular chitinase were isolated from soil and organic matter all over the Chonnam provincial area. The chemical composition, morphological, cultural and physiological properties of isolated strain S-25, S-42, S-172 and S-267 were studied in relation to the toxonomical properties. All of strains contained phospholipids such as phosphatidyl ethanolamine, phosphatidyl inositol and diphosphatidyl glycerol. The components of cell wall in all strains have L, L-DAP, Glutamic acid, Alanine, Glycine and Glucosamine. The surface of spore is smooth and colony is grey in all strains, Based on the results obtained in these experiments all of strains are identified as Streptomyces sp.S-25, Streptomyces sp.S-42, Streptomyces sp.S-l72 and Streptomyces sp. S-267.

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길항미생물 Pseudomonas sp. GRC3의 식물병원균에 대한 항균력

  • O, Ji-Hun;Park, Sin;Gang, Seon-Cheol
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.660-661
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    • 2001
  • Pseudomonas sp. GRC3 produced extracellular chitinase(s) and ${\beta}$-1,3-glucanase(s), possible biocontrol agents. Both of enzymes appeared to inhibit the growth of plant phathogens, especially Phytophthora capsici. Antifungal activities of Pseudomonas sp. GRC3 determined was more than 78% inhibition rate against P. capsici.

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Optimal Culture Condition and Extracellular Enzyme Activity of Metarhizium anisopliae (녹강균(Metarhizium anisopliae)의 최적 배양조건 및 효소활성)

  • 박영일;한영환
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.97-102
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    • 2000
  • In order to use Metarhizizmn~ anisopliae as a biological pesticide, effect of envrionmental factors on nlycelial growth, spore formation, and extracellular enzyme activity in culture broth of M. anisopliae DGUM 35001 was determined. Optimal temperature was 26^{\circ}C.$ and optimal pH ranged from 5 to 9. Among the complex media tested, MCM and SDPY media were the most favorable for mycelial growth. When Czapek-Dox agar was used as a mnimal medium, glucose and sucrose among the saccharides were very excellent source of carbohydrate. Among the biopolyners tested. chitin was the most favorable source for mycelial growth and produced high aerial inycelia. Urea and ammonium phosphate as an inorganic nitrogen source and bacto-peptone and soytone as an organic nitrogen source enhanced the mycelial growth When serine as a source of amino acid was supplemented, excellent mycelial growth was shown. Large amount of spores could be obtained from the aerial mycelia of starch medium. When the culture broth was filtrated and then the concentrate with ammonium sulfate was used as a crnde enzyme solution, high enzyme activities of amylase and protease were shown. However, lipase and chitinase activities were comparatively low.

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Purification and Properties of Chitosanase from Chitinolytic $\beta$-Proteobacterium KNU3

  • Yi, Jae-Hyoung;Jang, Hong-Ki;Lee, Sang-Jae;Lee, Keun-Eok;Choi, Shin-Geon
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.337-343
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    • 2004
  • A bacterial strain concurrently producing extracellular chitosanase and chitinase was isolated from soil and identified as a member of the $\beta$-subgroup of Proteobacteria through its 16S rRNA analysis and some biochemical analyses. The newly discovered strain, named as KNU3, had 99% homology of its 16S rRNA sequence with chitinolytic $\beta$-Proteobacterium CTE108. Strain KNU3 produced 34 kDa of chitosanase in addition to two chitinases of 68 kDa and 30 kDa, respectively. The purified chitosanase protein (ChoK) showed activity toward soluble, colloidal, and glycol chitosan, but did not exhibit any activity toward colloidal chitin. The optimum pH and temperature of ChoK were 6.0 and $70^{\circ}C$, respectively. The chitosanase was stable in the pH 4.0 to 8.0 range at $70^{\circ}C$, while enzyme activity was relatively stable at below $45^{\circ}C$. MALDI-TOF MS and N-terminal amino acid sequence analyses indicated that ChoK protein is related to chitosanases from Matsuebacter sp. and Sphingobacterium multivorum. HPLC analysis of chitosan lysates revealed that glucosamine tetramers and hexamers were the major products of hydrolysis.

Antifungal Activity of Serratia marcescens Culture Extracts against Phytopathogenic Fungi: Possibility for the Chitinases Role

  • Cho, Moo-Je;Lee, Sang-Yeol;Gal, Sang-Wan;Hwang, Jae-Ryoung;Yoon, Hae-Won
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.209-214
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    • 1992
  • Serratia marcescens co-cultured with various phytopathogenic fungi, including Rhizopus stolonifer, Helminthosporium allii, Pyricularia oryzae, Fusarium oxysporium and Collectothricom cassiicola, in an LB- agar medium containing 1.5% swollen chitin, significantly inhibitied fungal growth. Fungal hyphae grew rapidly outward from the culture dish center, but the hyphal extensions of the pathogenic fungi were significantly inhibited in a perimetric contact area with S. marcescens. This was especially evident in pathogenic fungi which have a high chitin content in their cell walls. The extracellular chitinase activities of S. marcescens were increased seven fold by the addition of 1.5% swollen chitin to the LB-broth, compared to chitinase activities in a culture medium without chitin. The type of induction was dependent on the various forms of chitin used. When the culture supernatant of S. marcescens or the chitinases of Streptomyces griceus purchased from Sigma Chemical Co., were incubated with the mycelium of F. oxysporium, the mycelium gradually burst as cultivation time progressed and completely lysed after incubation for 2 days. On the other hand, E. coli extract did not hydrolyze the F. oxysporium mycelium at all. These data showed that the chitinolytic activities of S. marcescens play important roles in the biochemical control of phytopathogenic fungi.

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Diurnal Fluctuations of Saprophytic Bacterial distribution and Their Extracellular Enzyme Activities in the Overlying Waters of Sediment of the Yellow Sea near Daesan, Korea (대산인근 해역에서 간만조에 따른 종속영양세균의 일일 분포와 세포외 효소 활성력의 변화)

  • Lee, Geon-Hyoung;Gang-Guk Choi;Chun-Bong Baek
    • The Korean Journal of Ecology
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    • v.18 no.3
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    • pp.409-418
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    • 1995
  • As a part of studying the function and structure of the mudflat environment of the Yellow Sea, seawater samples in the overlying waters of sediment near Daesan were collected every hour on March 29 (spring tides) and on April 5 (neap tides), 1995 to study the diurnal distribution of aerobic saprophytic bacteria and their extracellular enzyme activities. The diurnal distribution of aerobic saprophytic bacteria ranged from 1.0 X $10^{2}$ to 7.07 X $10^{3}$ cfu /ml at spring tides and from 1.0 X $10^{2}$ to 8.3 X $10^{3}$ cfu /ml at neap tides. The diurnal variations of aerobic saprophytes at the suface waters were greater than those of middle and bottom waters. However, th diurnal fluctuation of saprophyte numbers at spring tides showed no significant difference compared with that at neap tides. The numbers of three physiological groups of aerobic hacteria (proteolytic, lipolytic and amylolytic bacteria) at the surface waters during spring and neap tides were lower than those at the middles and bottom waters. The diurnal variations of five extracellular enzyme activities at the surface waters during the survey period showed lower values than those at the middle and botton waters. Among the measured extracellular enzyme activities, phosphatase showed the highest. However, the activities of amylase, chitinase and cellulase showed a similar tendency.

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Expression of pqq Genes from Serratia marcescens W1 in Escherichia coli Inhibits the Growth of Phytopathogenic Fungi

  • Kim, Yong-Hwan;Kim, Chul-Hong;Han, Song-Hee;Kang, Beom-Ryong;Cho, Song-Mi;Lee, Myung-Chul;Kim, Young-Cheol
    • The Plant Pathology Journal
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    • v.22 no.4
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    • pp.323-328
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    • 2006
  • Serratia marcescens W1, isolated from cucumber-cultivated soil in Suwon, Korea, evidenced profound antifungal activity and produced the extracellular hydrolytic enzymes, chitinase and protease. In order to isolate the antifungal genes from S. marcescens W1, a cosmid genomic library was constructed and expressed in Escherichia coli. Transformants exhibiting chitinase and protease expression were selected, as well as those transformants evidencing antifungal effects against the rice blast fungus, Magnaporthe grisea, and the cucumber leaf spot fungus, Cercospora citrullina. Cosmid clones expressing chitinase or protease exerted no inhibitory effects against the growth of fungal pathogens. However, two cosmid clones evidencing profound antifungal activities were selected for further characterization. An 8.2 kb HindIII fragment from these clones conditioned the expression of antagonistic activity, and harbored seven predicted complete open reading frames(ORFs) and two incomplete ORFs. The deduced amino acid sequences indicated that six ORFs were highly homologous with genes from S. marcescens generating pyrroloquinoline quinone(PQQ). Only subclones harboring the full set of pqq genes were shown to solubilize insoluble phosphate and inhibit fungal pathogen growth. The results of this study indicate that the functional expression of the pqq genes of S. marcescens W1 in E. coli may be involved in antifungal activity, via as-yet unknown mechanisms.

Antagonistic Potential of Native Trichoderma viride Strain against Potent Tea Fungal Pathogens in North East India

  • Naglot, A.;Goswami, S.;Rahman, I.;Shrimali, D.D.;Yadav, Kamlesh K.;Gupta, Vikas K.;Rabha, Aprana Jyoti;Gogoi, H.K.;Veer, Vijay
    • The Plant Pathology Journal
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    • v.31 no.3
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    • pp.278-289
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    • 2015
  • Indigenous strains of Trichoderma species isolated from rhizosphere soils of Tea gardens of Assam, north eastern state of India were assessed for in vitro antagonism against two important tea fungal pathogens namely Pestalotia theae and Fusarium solani. A potent antagonist against both tea pathogenic fungi, designated as SDRLIN1, was selected and identified as Trichoderma viride. The strain also showed substantial antifungal activity against five standard phytopathogenic fungi. Culture filtrate collected from stationary growth phase of the antagonist demonstrated a significantly higher degree of inhibitory activity against all the test fungi, demonstrating the presence of an optimal blend of extracellular antifungal metabolites. Moreover, quantitative enzyme assay of exponential and stationary culture filtrates revealed that the activity of cellulase, ${\beta}$-1,3-glucanase, pectinase, and amylase was highest in the exponential phase, whereas the activity of proteases and chitinase was noted highest in the stationary phase. Morphological changes such as hyphal swelling and distortion were also observed in the fungal pathogen grown on potato dextrose agar containing stationary phase culture filtrate. Moreover, the antifungal activity of the filtrate was significantly reduced but not entirely after heat or proteinase K treatment, demonstrating substantial role of certain unknown thermostable antifungal compound(s) in the inhibitory activity.

Characterization of L-asparaginase-producing Trichoderma spp. Isolated from Marine Environments

  • Woon-Jong, Yu;Dawoon, Chung;Yong Min, Kwon;Seung Sub, Bae;Eun-Seo, Cho;Hye Suck, An;Grace, Choi
    • Journal of Marine Life Science
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    • v.7 no.2
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    • pp.121-128
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    • 2022
  • L-asparaginase (ASNase) is a therapeutic enzyme used to treat acute lymphoblastic leukemia. Currently, the most widely used ASNases are originated from bacteria. However, owing to the adverse effects of bacterial ASNases, new resources for ASNase production should be explored. Fungal enzymes are considered efficient and compatible resources of natural products for diverse applications. In particular, fungal species belonging to the genus Trichoderma are well-known producers of several commercial enzymes including cellulase, chitinase, and xylanase. However, enzyme production by marine-derived Trichoderma spp. remains to be elucidated. While screening for extracellular ASNase-producing fungi from marine environments, we found four strains showing extracellular ASNase activity. Based on the morphological and phylogenetic analyses using sequences of translation elongation factor 1-alpha (tef1α), the Trichoderma isolates were identified as T. afroharzianum, T. asperellem, T. citrinoviride, and Trichoderma sp. 1. All four strains showed different ASNase activities depending on the carbon sources. T. asperellem MABIK FU00000795 showed the highest ASNase value with lactose as a carbon source. Based on our findings, we propose that marine-derived Trichoderma spp. are potential candidates for novel ASNase production.

Isolation and Morphological Characterization of Ttichoderma harzianum SJG-99721, a Powerful Biocontrol Agent (길항작용을 나타내는 Trichoderma harzianum SJG-99721의 분리 및 형태학적 특징)

  • 이호용;민봉희
    • Korean Journal of Environmental Biology
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    • v.20 no.2
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    • pp.130-135
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    • 2002
  • Species of Genus Trichoderma are commercially applied as biological control agents against fungal Pathogens. A powerful biocontrol agent, Trichoderma sp. SJG-99721 was isolated from 305 isolates by morphological characters, chitinase activities and antifungal activities against Phytophthora capsiei. The isolate was identified as Trichoderma harzianum from various features such as growth rate at $27^\circ{C}$, significant growth ratio of $27^\circ{C}$ to $17^\circ{C}$, amount of aerial mycelium, types of branching: system, and disposition patterns of phialide and phialospore. Trichoderma harzianum SJG-99721 have been shown to act as a powerful biological agent against fungal phytopathogens; Botrytis cinerea, Rhizoctonia solani, Phytophthora cryptogea, Phytophthora capsiei, Sclerotinia sclerotiorum, Mycoshaerella melonis, Alternaria sotani, Fusarium oxysporum, Collectotrichum gloesporioodes, Alternaria alternata, Phythium ultimum, Phytophthora drechsleri, Pyricularia grisea.