• Title/Summary/Keyword: chitinase production

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Antifungal Mechanism of Pseudomonas stutzeri YPL-l for Biocontrol of Fusarium solani causing Plant Root Rot (식물근부균 Fusarium solani에 대한 Pseudomonas stutzeri YPL-1의 생물학적 방제기작)

  • 임호성;김상달
    • Microbiology and Biotechnology Letters
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    • v.18 no.1
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    • pp.81-88
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    • 1990
  • For the selection of powerful antagonistic bacterium for biological control of soilborne Fusarium solani causing root rot of many important crops, the best YPL-1 strain was selected among 300 strains of bacteria isolated from rhizosphere in ginseng root rot-suppressive soil. The strain was identified to be a species to Pseudomonas stutzeri. With in vitro fungal inhibition tests, antagonistic substance of P. stutzeri YPL-1 against F. solani was presumed to be heat unstable, macromolecular substances such as protein. Also, it was shown that antifungal activity of P. stutzeri YPL-1 increased in proportion to its chitinase production. P. stutzeri YPL-M122 (chi-, lam -) which was deprived of the productivity of chitinase and laminarinase by NTG mutagenesis had lost antifungal activity, completely. And P. stutzeri YPL-MI53 (chi-) had only 4.1% of its antifungal activity. P. stutzeri YPL-1 was not able to produce any extracellular siderophore in iron-deficent minimal medium. It is confident that the antifungal mechanism of P. stutzeri YPL-1 for biocontrol of F. solani depends on lysis rather than antibiosis :the mechanism of lysis appears to involve enzymatic degradation of the cell will components of F. solani by hydrolytic enzymes of more chitinase and less laminarinase.

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Isolation and Characterization of Chitinoloytic Strain, Bacillus atrophaeus CJ-3. (전통발효식품으로부터 Chitin 분해 미생물의 분리 및 특성 규명)

  • 고보경;최인순;이상현;임채오;이성호;갈상완;최영주
    • Journal of Life Science
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    • v.14 no.3
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    • pp.501-508
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    • 2004
  • A bacterial strain CJ-3 which produced chitinase was isolated from Korean traditional soy sauce. Using 16S rDNA analysis, the strain CJ-3 was identified as Bacillus atrophaeus. The approximate molecular weight of the putative chitinase enzyme was 31.0 kDa and the enzyme activity was remarkably induced by addition of colloidal chitin (0.5, 1.0, 2.0%). The antioxidant activity was increased 53% by the browning reaction products of B. atrophneus CJ-3. Escherichia. coli lipopolysaccharides (LPS)-induced production of nitric oxide(NO) was reduced up to 45% by the browning reaction product in RAW264.7 macrophage. Inhibition of cell viability in the presence of LPS was recovered to normal level by the browning reaction product. These results suggest that browning reaction of B. atrophaeus CJ-3 plays an important role for activation of immune system. B. atrophaeus CJ-3 exhibited optimum temperature and pH of 37$^{\circ}C$ and pH 7.0∼8.0, respectively. The major intracelluar free amino acid was determined to be glutamate.

Chitinase을 생산하는 곤충병원미생물 Metarhizium anisopliae HY-2(KCTC 0156BP)의 토양해충 생물검정

  • Seo, Eun-Yeong;Son, Gwang-Hui;Sin, Dong-Ha;Kim, Gi-Deok;Park, Du-Sang;Park, Ho-Yong
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.469-472
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    • 2002
  • Solid state fermentation was performed for the production of entomopathogenic fungus Metarhizium anisopliae HY-2 using wheat bran media containing rice bran. Fungal growth in a solid state fermentation system was estimated by viable cell count, spore count, and mycelial biomass. It was used chemical method measuring N-acetyl-glucosamine (chitin) content for estimating of mycelial biomass. In static flask culture, viable cell reached 2.40 ${\times}$ $10^8$ cfu/g at 23 days of culture at $27^{\circ}C$ and then mycelial biomass was 41.59 mg/g. Specific growth rate(${\mu}$ max) was 0.0418 $h^{-1}$ between 3 and 9 days when estimated by viable cell count and was 0.00976 $h^{-1}$ between 9 and 17 days when N-acetylglucosamine content was measured. Viable cells reached 1.12 ${\times}$ $10^8$ cfu/g in polypropylene-bag at 28 days of culture at $27^{\circ}C$. Formulated microbial pesticide containing M. anisopliae HY-2 were tested their bio-activity against Chestnut Brown Chafer (Adoretus tenuimaculatus). The protection rate of the liquid culture showed 13 ${\sim}$ 26 % with 1st to 3rd instar, and spore suspension of M. anisopliae HY-2 showed 56 ${\sim}$ 64%. Conidia produced by large scale solid-state fermentation showed 20 ${\sim}$ 27 % activity 60 ${\sim}$ 64 % with M. anisopliae HY-2.

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Optimization of Medium Components for the Production of Antagonistic Lytic Enzymes Against Phytopathogenic Fungi and Their Biocontrol Potential

  • Lee, Yong Seong;Neung, Saophuong;Park, Yun Suk;Kim, Kil Yong
    • Korean Journal of Soil Science and Fertilizer
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    • v.47 no.4
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    • pp.299-305
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    • 2014
  • In this paper, fractional factorial screening design (FFSD) and central composition design (CCD) were used to optimize the medium components for producing chitinase and gelatinase by Lysobacter capsici YS1215. Crab shell powder, nutrient broth and gelatin were proved to have significant effects on chitinase and gelatinase activity by FFSD first. An optimal medium was obtained by using a three factor CCD, which consisted of nutrient broth of $2.0gL^{-1}$, crab shell powder of $2.0gL^{-1}$ and gelatin of $1.0gL^{-1}$, respectively with the highest chitinase activity ($3.34UmL^{-1}$) and gelatinase activity ($14.15UmL^{-1}$). This value was 3.76 and 1.11 fold of the chitinase and gelatinase activity, respectively, compared to the lowest productive medium in the design matrix. In investigating potential of these enzymes partially purified from L. capsici YS1215 for biotechnological use, the crude enzymes was found to be inhibition against pathogenic fungal mycelia: Colletotrichum gleosporioides, Phytophthora capsici, and Rhizoctonia solani. In this study, we demonstrated the optimal medium for producing the chitinolytic and gelatinolytic enzymes by the strain YS1215 and the role of their enzymes that may be useful for further development of a biotechnological use and agricultural use for biological control of phytopathogenic fungi.

Effectiveness of Various Pseudomonas spp. and Burkholderia caryophylli Containing ACC-Deaminase for Improving Growth and Yield of Wheat (Triticum aestivum L.)

  • Shaharoona, B.;Jamro, G.M.;Zahir, Z.A.;Arshad, M.;Memon, K.S.
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1300-1307
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    • 2007
  • This study assessed the possible role of different traits in selected plant growth-promoting rhizobacteria (PGPR) for improving wheat growth and yield under natural conditions. Rhizobacteria exhibiting 1-aminocyclopropane-1-carboxylate (ACC)-deaminase activity were isolated and screened for their growth-promoting activity in wheat under axenic conditions. Five isolates belonging to Pseudomonas and one Burkholderia caryophylli isolate that showed promising performances under axenic conditions were selected and characterized for in vitro ACC-deaminase activity, chitinase activity, auxin production, P solubilization, and root colonization. These isolates were then used as inocula for wheat cultivated under natural conditions in pot and/or field trials. Significant increases in root elongation, root weight, tillers per pot, 1,000-grain weight, and grain and straw yields were observed in response to inoculation with PGPR in the pot trials. Inoculation with these PGPR was also effective under field conditions and increased the wheat growth and yield significantly. However, the efficacy of the strains was inconsistent under the axenic, pot, and field conditions. Pseudomonas fluorescens ($ACC_{50}$), which exhibited a relatively high in vitro ACC-deaminase activity, chitinase activity, auxin production, and P solubilization and more intensive root colonization, was the most efficient isolate under the field conditions. Therefore, these results demonstrated that ACC-deaminase activity is an efficient parameter for the selection of promising PGPR under axenic conditions. However, additional traits of PGPR, including auxin production, chitinase activity, P solubilization, and root colonization, are also important for selecting PGPR as biofertilizers.

Chitinase Production and Isolation of Serratia plymuthica AL-1 Antagonistic to White Rot Fungi from Allium fistulosum Roots. (대파 뿌리로부터 흑색썩음균핵병균에 길항하는 Serratia plymuthica AL-1의 분리 및 Chitinase의 생산)

  • 주길재;이익희;김진호
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.135-141
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    • 2002
  • This study was carried out to isolate antagonistic bacterium against Sclerotium cepivorum causing Allium fistulosum white rot. Total of 146 strains were isolated from A. fistulosum roots. The isolates were screened for antagonism to S. cepivorum and the isolated strain No. AL-1 was selected among these bacteria. It was identified as Serratia plymuthica based on morphological and physiological characteristics according to the Bergey's mannual of systematic bacteriology and 16S rDNA sequences methods. Serratia plymuthica AL-1 showed broad spectrum of antifungal activities against plant pathogenic fungi Alternaria altrata, Colletotrichum gleosporioids, Phoma sp., Rhizoctonia solani, Sclerotinia sclerotiorum, Stemphylium solani, Fusarium oxysporium niveum but not inhibited Didymella bryoniae. When S. plymuthica AL-1 cultivated in the TSB medium containing 1% colloidal chitin, the high molecular fraction (>10 kDa) have chitinase activity (3.2 units/ml) and the low molecular fraction (<10 kDa) have not chitinase activity. Oppositely, after heat treatment (80℃ for 30 min) of the cultivation supernatant, the high molecular fractions have not antifungal activity but the low molecular fractions have antifungal activity.

Antifumgal Activity and Identification of an Actinomycetes Strain Isolated from Mummified Peaches (복숭아 미이라과로부터 분리한 방선균의 항균 활성 및 동정)

  • 임태헌;이정목;장태현;차병진
    • Microbiology and Biotechnology Letters
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    • v.28 no.3
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    • pp.161-166
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    • 2000
  • Antifungal Activity and Identification of an Actinomycetes Strain Isolated from Mummified Peaches. Lirn, Tae Heon*, Jung Mok Lee, Tae Hyun Chang, and Byeongjin Chal. *Research Institute of Plant Nutrient, Oaeyu Co, Inc. Kyongsan 712-820, Korea, 1 Department of Agricultural Bi%g'f Chungbul< NatJ"onal Univershy, Cheongju 367-763, Korea - An actinomycetes strain which produced chitinase, urease, and antifungal substances to MoniliniaJhtcticola was isolated from peaches mununified by Moniliniafructicola. The strain TH-04 was identified as Streptomyces sp. based on cultural and lTIOIphological characteristics, cell wall diaminopimelic acid, and sugar patterns ofwhole~cell extracts. Streptomyces sp. TH~04 showed antifungal activity to several fungi including Moniliniafructicola, Colletotrichum gloeosporioides, Magnaponhe grisea, Rhizoctonia solani, Phytophthora capsici, Altemaria kikuchiana, Fusarium solani, and Fusarium O),ysporum. The optimum cultural conditions for the production of antifungal substances were $20^{\circ}C$pH 7, and 7 days.

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Chitinase-producing Salinivibrio bacteria isolated from salt-fermented shrimp with antimicrobial and safety assessments

  • Le, Bao;Chung, Gyuhwa;Yang, Seung Hwan
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.233-238
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    • 2018
  • Chitinases are glycosyl hydrolases which cleave the ${\beta}$-1,4 linkage of chitin into oligo or monomers of N-acetylglucosamine. These bacterial enzymes have been used for a wide range of applications in the food and pharmaceutical industries. In this study, we isolated two potential chitinolytic strains, BAO-01 and BAO-02, from salt-fermented shrimp, which were shown to belong to the genus Salinivibrio through genetic characterization using 16S rRNA. These isolates were gram-positive, rod-shaped, and non-spore forming. BAO-01 showed greater growth and chitinase activity than BAO-02 after the incubation at $37^{\circ}C$ for 4 days. Both strains grew on a wide range of carbon and nitrogen sources, pH values, temperatures, and salt levels. However, they showed minor biochemical differences. In addition, their antimicrobial activities against foodborne pathogens and antibiotic susceptibilities were evaluated. These Salinivibrio spp. did not show bioamine production, hemolytic activity, and mucin degradation. Therefore, the in vitro screening results suggested that these bacteria could be widely used as new candidates for chitin hydrolyzation and seafood fermentation.

Characterization and Antifungal Activity from Soilborne Streptomyces sp. AM50 towards Major Plant Pathogens

  • Jang, Jong-Ok;Lee, Jung-Bok;Kim, Beam-Soo;Kang, Sun-Chul;Hwang, Cher-Won;Shin, Kee-Sun;Kwon, Gi-Seok
    • Korean Journal of Environmental Agriculture
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    • v.30 no.3
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    • pp.346-356
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    • 2011
  • BACKGROUND: Chemical fungicides not only may pollute the ecosystem but also can be environmentally hazardous, as the chemicals accumulate in soil. Biological control is a frequently-used environment-friendly alternative to chemical pesticides in phytopathogen management. However, the use of microbial products as fungicides has limitations. This study isolated and characterized a three-antifungal-enzyme (chitinase, cellulase, and ${\beta}$-1,3-glucanase)-producing bacterium, and examined the conditions required to optimize the production of the antifungal enzymes. METHOD AND RESULTS: The antifungal enzymes chitinase, cellulase, and ${\beta}$-1,3-glucanase were produced by bacteria isolated from an sawmill in Korea. Based on the 16S ribosomal DNA sequence analysis, the bacterial strain AM50 was identical to Streptomyces sp. And their antifungal activity was optimized when Streptomyces sp. AM50 was grown aerobically in a medium composed of 0.4% chitin, 0.4% starch, 0.2% ammonium sulfate, 0.11% $Na_2HPO_4$, 0.07% $KH_2PO_4$, 0.0001% $MgSO_4$, and 0.0001% $MnSO_4$ at $30^{\circ}C$. A culture broth of Streptomyces sp. AM50 showed antifungal activity towards the hyphae of plant pathogenic fungi, including hyphae swelling and lysis in P. capsici, factors that may contribute to its suppression of plant pathogenic fungi. CONCLUSION(S): This study demonstrated the multiantifungal enzyme production by Streptomyces sp. AM50 for the biological control of major plant pathogens. Further studies will investigate the synergistic effect, to the growth regulations by biogenic amines and antifungal enzyme gene promoter.