• Title/Summary/Keyword: extracellular protease production

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A two-component sensor kinase (GacS) mediated signal transduction pathway involved in production of antifungal compounds in Pseudomonas chlororaphis O6.

  • Kang, Beom-Ryong;Lee, Jung-Hoon;Kim, Hyun-Jung;Cho, Baik-Ho;Kim, Young-Cheol
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.99.1-100
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    • 2003
  • E. intermedium Blocontrol activity of a P. chlororaphis rhizobacteium O6, depends to the synthesis of extracellular secondary metabolites and exoenzymes, thought to antagonize the pathogenicity of a variety of phytopathogenic fungi. The production of secondary metabolites and exoenzymes in O6, depends essentially on the GacS-mediated signal transduction pathway, which activates largely unknown signal transduction pathway. To exploit the GacS-mediated signal transdcution pathway involved in activation of ph genes that are necessary for biosynthesis of phenazine from P. chlororaphis O6, we cloned and sequenced the phz operon, rpoS gene encoding stationary specific sigma factor, ppx gene encoding polyphosphatase, and lon gene encoding ion protease. Expression of each gene in wild type and GacS mutant were analyzed by RT-PCR. Transcripts from rpoS, phzI enconing acylhomoserine lactone (AHL) synthase, and ph structural genes in the GacS mutant were reduced in each of these growth phases compared to the wild type. The GacS or Lon mutant was found to be deficient in the production of phenzines, exoenzymes, and the acylhomoserine lactone. These mutants were not complemented by ph operon and addition of exogenous AHL. These results indicate that the GacS global regulatory systems controls phenazine production at multiple levels. Future research will focus to identifying the GacS-mediated regulatory cascade involving in production of phenazine in P. chlororaphis.

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Isolation and Characterization of Airborne Mushroom Damaging Trichoderma spp. from Indoor Air of Cultivation Houses Used for Oak Wood Mushroom Production Using Sawdust Media

  • Kim, Jun Young;Kwon, Hyuk Woo;Lee, Dong Hyeung;Ko, Han Kyu;Kim, Seong Hwan
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.674-683
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    • 2019
  • Some species of the Trichoderma genus are reported as the major problem in oak wood mushroom production in Korea. In spite of economic loss by the fungi, scientific information on airborne Trichoderma species is not much available. To generate information for disease management development we analyzed airborne Trichoderma. A total of 1,063 fungal isolates were purely obtained from indoor air sampling of cultivation houses used for oak wood mushroom using sawdust media. Among the obtained isolates, 248 isolates were identified as Trichoderma fungi including T. harzianum, T. atroviride, T. citrinoviride, and T. pseudokoningii, by morphological and molecular analysis. T. harzianum was dominant among the four identified species. All the four Trichoderma species grew fast on solid nutrient media tested (potato dextrose agar [PDA], malt extract agar [MEA], Czapek's Dox + yeast extract agar [CYA] and cornmeal dextrose agar). Compact mycelia growth and mass spore production were better on PDA and CYA. In addition, T. harzianum and T. citrinoviride formed greenish and yellowish mycelium and spores on PDA and CYA. Greenish and yellowish pigment was saturated into PDA only by T. pseudokoningii. These four Trichoderma species could produce extracellular enzymes of sawdust substrate degradation such as β-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease. Their mycelia inhibited the growth of oak wood mushroom mycelia of two tested cultivars on dual culture assay. Among of eleven antifungal agents tested, benomyl was the best to inhibit the growth of the four Trichoderma species. Our results demonstrate that the airborne Trichoderma fungi need to be properly managed in the cultivation houses for safe mushroom production.

Isolation, Characterization, and Use for Plant Growth Promotion Under Salt Stress, of ACC Deaminase-Producing Halotolerant Bacteria Derived from Coastal Soil

  • Siddikee, M.A.;Chauhan, P.S.;Anandham, R.;Han, Gwang-Hyun;Sa, Tong-Min
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1577-1584
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    • 2010
  • In total, 140 halotolerant bacterial strains were isolated from both the soil of barren fields and the rhizosphere of six naturally growing halophytic plants in the vicinity of the Yellow Sea, near the city of Incheon in the Republic of Korea. All of these strains were characterized for multiple plant growth promoting traits, such as the production of indole acetic acid (IAA), nitrogen fixation, phosphorus (P) and zinc (Zn) solubilization, thiosulfate ($S_2O_3$) oxidation, the production of ammonia ($NH_3$), and the production of extracellular hydrolytic enzymes such as protease, chitinase, pectinase, cellulase, and lipase under in vitro conditions. From the original 140 strains tested, on the basis of the latter tests for plant growth promotional activity, 36 were selected for further examination. These 36 halotolerant bacterial strains were then tested for 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity. Twenty-five of these were found to be positive, and to be exhibiting significantly varying levels of activity. 16S rRNA gene sequencing analyses of the 36 halotolerant strains showed that they belong to 10 different bacterial genera: Bacillus, Brevibacterium, Planococcus, Zhihengliuella, Halomonas, Exiguobacterium, Oceanimonas, Corynebacterium, Arthrobacter, and Micrococcus. Inoculation of the 14 halotolerant bacterial strains to ameliorate salt stress (150 mM NaCl) in canola plants produced an increase in root length of between 5.2% and 47.8%, and dry weight of between 16.2% and 43%, in comparison with the uninoculated positive controls. In particular, three of the bacteria, Brevibacterium epidermidis RS15, Micrococcus yunnanensis RS222, and Bacillus aryabhattai RS341, all showed more than 40% increase in root elongation and dry weight when compared with uninoculated salt-stressed canola seedlings. These results indicate that certain halotolerant bacteria, isolated from coastal soils, have a real potential to enhance plant growth under saline stress, through the reduction of ethylene production via ACC deaminase activity.

Isolation and Characterization of Transposon \ulcorner¨ªKm-Mediated Nonpathogenic Mutants of Xanthomonas campestris pv. vesicatoria (고추 세균성 반점병균의 비병원성 돌연변이체 분리 및 생리적 특성)

  • 윤영채;김용식;조용섭
    • Korean Journal Plant Pathology
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    • v.11 no.3
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    • pp.265-270
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    • 1995
  • Transposon mutation of Xanthomonas campestris pv. vesicatoria (Xcv) was induced by using transposon omegon ($\Omega$)-Km (Tn $\Omega$Km), which was confirmed by resistance to kanamycin (KMr), and nonpathogenic mutants were selected through the inoculation test on pepper plants. The mutagenesis frequency was about 6$\times$10-8, and 53 out of 2,000 Kmr bacterial colonies tested were nonpathogenic to the pepper cultivar Cheung-Hong. Optimum conditions for the Tn $\Omega$Km mutagenesis of Xcv were Luria Bertani (LB) broth medium for culture of Xcv, yeast extract-dextrose-CaCO3 (YDC) agar medium for selection of Tn $\Omega$Km-mediated mutants, and over 1 to 2 in the ratio of the donor (Escherichia coli S17-1 with the plasmid pJFF350 $\Omega$Km) and the recipient (Xcv) in the culture for the mutagenesis. One of the 4 nonpathogenic mutants (WNP1, WNP3, WNP4 and WNP5), which had been reconfirmed through the inoculation on pepper cv. Dabokgun, showed no differences in the production of exoenzymes such as protease and polygalacturonase and extracellular polysaccharides in vitro and the bacterial growth rate from those of the wild type of Xcv.

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Studies on Exudative Epidermitis in Pigs : I. Isolation and Some Properties of Staphylococcus hyicus subsp. hyicus from Diseased and Healthy Pigs (돼지 삼출성(渗出性) 표피염(表皮炎)에 관한 연구(硏究) : I. 발증돈(發症豚) 및 건강돈(健康豚)으로부터 Staphylococcus hyicus subsp. hyicus의 분리(分離) 및 그 성장(性狀))

  • Park, Cheong-kyu;Kang, Byong-kyu
    • Korean Journal of Veterinary Research
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    • v.26 no.2
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    • pp.251-257
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    • 1986
  • Isolation of Staphylococcus hyicus subsp. hyicus from piglets suffering from exudative epidermitis and healthy pigs was performed, and some properties of the isolates were examined. Twentry-six litters of exudative epidermitis observed in the field were sucking piglets with ages ranging from 1 to 5 weeks and most (73.1%) of these cases occurred in piglets between 2 and 3 weeks of age. Staphylococcus hyicus subsp. hyicus was isolated front 192(38.9%) of 494 healthy pigs. The rate of isolation of these organism from healthy pigs was found to vary greatly among farms at an isolation rate of 33.3% to 45.5% and this organism was isolated more frequently front sucking piglets below the age of 8 weeks than adult pigs. All of the isolates originated from the diseased and healthy pigs were positive in heat-stable DNase, Tween 80 hydrolysis, gelatinase, protease, but negative in clumping factor and hemelysin on sheep blood agar. There was no difference related to origin in the production of extracellular active substances. Typical lesions of the classic exudative epidermitis were produced by inoculation of Staphylococcus hyicus subsp. hyicus isolated from the diseased and healthy pigs. The pathogenic potentials of isolates from healthy pigs were no different from that of the isolates obtained from diseased pigs.

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Isolation of new microorganisms which degrades the protein of a food garbage efficiently and its application (음식물 쓰레기중의 단백질을 효과적으로 분해하는 신규 미생물의 분리 및 응용)

  • Koo Kyung-Wan;Chung Yong-Hyun;Hong Sung-Hee;Oh Sang-Hoon;Kim Dong-seop;Jeon Hee-Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.6 no.4
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    • pp.342-348
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    • 2005
  • In this study, novel strains showing better protein degradation activity were isolated for the production of effective compost from garbages. Well growing bacteria with clear zone on the skim milk agar media were isolated . The strain was identified as Bacillus subtilis PNV-1 through various biochemical tests, Bergy's manual of determinative bacteriology and 165 rDNA partial sequence. The extracellular protease of the strain PNV-1 has its activity at broad pH and the optimal temperature was $50^{\circ}C$. Also, the strain PNV-1 was highly tolerant to high concentration of salt, red/black pepper and mustard.

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Potato Soft Rot Caused by Psychrotolerant Pseudomonas sp. from Subarctic Tundra Soil

  • Sungho Woo;Yung Mi Lee;Dockyu Kim
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.399-404
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    • 2023
  • Agricultural activities and the number of farms in the subarctic regions have been increasing annually after the coronavirus disease 2019 pandemic to achieve food self-sufficiency. Potatoes are vulnerable to soft rot bacteria at all stages of production, storage, and transportation. A novel bacterium, Pseudomonas sp. N3-W, isolated from Alaska tundra soil, grows at 5-25℃ and produces extracellular protease(s). N3-W caused necrotic spots (hypersensitivity) in hot pepper leaves and soft rot disease (pathogenicity) in potato tubers. The psychrotolerant N3-W caused significant soft rot symptoms on potatoes at a broad temperature range (5℃, 15℃, and 25℃). In contrast, mesophilic Pectobacterium carotovorum KACC 16999 induced severe rotting symptoms in potatoes at their optimal growth temperature of 15℃ and 25℃. However, it barely produced symptoms at 5℃, which is the appropriate storage and transportation temperature for potatoes. The results of pathogenicity testing imply that psychrotolerant soft rot pathogens from polar regions may cause severe soft rot not only during the crop growing season but also during storage and transportation. Our study indicates the possibility of new plant pathogen emergence and transmission due to the expansion of crop cultivation areas caused by permafrost thawing in response to recent polar warming.

A Study of the Diversity and Profile for Extracellular Enzyme Production of Aerobically Cultured Bacteria in the Gut of Muraenesox cinereus (갯장어(Muraenesox cinereus) 장으로부터 호기적 조건에서 분리된 미생물의 다양성 및 세포외 효소 생산능 분석에 관한 연구)

  • Lee, Yong-Jik;Oh, Do-Kyoung;Kim, Hye Won;Nam, Gae-Won;Sohn, Jae Hak;Lee, Han-Seung;Shin, Kee-Sun;Lee, Sang-Jae
    • Journal of Life Science
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    • v.29 no.2
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    • pp.248-255
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    • 2019
  • This research confirmed the diversity and characterization of gut microorganisms isolated from the intestinal organs of Muraenesox cinereus, collected on the Samcheonpo Coast and Seocheon Coast in South Korea. To isolate strains, Marine agar medium was basically used and cultivated at $37^{\circ}C$ and pH7 for several days aerobically. After single colony isolation, totally 49 pure single-colonies were isolated and phylogenetic analysis was carried out based on the result of 16S rRNA gene DNA sequencing, indicating that isolated strains were divided into 3 phyla, 13 families, 15 genera, 34 species and 49 strains. Proteobacteria phylum, the main phyletic group, comprised 83.7% with 8 families, 8 genera and 26 species of Aeromonadaceae, Pseudoalteromonadaceae, Shewanellaceae, Enterobacteriaceae, Morganellaceae, Moraxellaceae, Pseudomonadaceae, and Vibrionaceae. To confirm whether isolated strain can produce industrially useful enzyme or not, amylase, lipase, and protease enzyme assays were performed individually, showing that 39 strains possessed at least one enzyme activity. Especially the Aeromonas sp. strains showed all enzyme activity tested. This result indicated that isolated strains have shown the possibility of the industrial application. Therefore, this study has contributed for securing domestic genetic resources and the expansion of scientific knowledge of the gut microbial community in Muraenesox cinereus of South Korea.

The Optimal Condition and Enzyme Activity of Entomopathogenic Fungus Beauveria bassiana Using Extracted Rice Bran (미강추출물을 이용한 곤충병원성 곰팡이 Beauveria bassiana의 최적 배양조건 및 효소활성)

  • Kim, Chang-Su;Lee, Jung-Bok;Kim, Beam-Soo;Lee, Min-Hye;Kang, Kyeong-Muk;Joo, Woo-Hong;Kim, Jin-Won;Im, Dae-Joon;Kwon, Gi-Seok
    • Journal of Life Science
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    • v.23 no.8
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    • pp.1010-1018
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    • 2013
  • The greenhouse whitefly, Bemisia tabaci, is considered one of the most destructive pests of crops. In this study, we aimed to determine the optimal liquid culture conditions in shake flasks for maximal sporulation of Beauveria bassiana M130 using rice bran. The optimal initial pH for the spore production of B. bassiana using extracted rice bran medium was 5.2 and $28^{\circ}C$. The screening in shake flasks of carbon and nitrogen sources resulted in the identification of an optimal medium based on 0.5% $(NH_4)_2SO_4$, with extracted rice bran 8:1. Using this medium, a production level of $2.15{\times}10^9$ spores per ml was obtained after six days from culture inoculation at $28^{\circ}C$ in a rotary shaking incubator at 130 rpm. In addition, the specific activities of extracellular enzymes of chitinase and protease were $4,296{\mu}mol$ and $375{\mu}mol$, respectively. These results suggest that Beauveria bassiana M130 could be a bio-controller for the greenhouse whitefly.

Optimal Culture Conditions on the Keratinase Production by Bacillus sp. SH-517. (Bacillus sp. SH-517에 의한 keratinase의 생성 최적 배양 조건)

  • Bang, Byung-Ho;Rhee, Moon-Soo;Lim, Ki-Hwan;Yi, Dong-Heui
    • Journal of Life Science
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    • v.18 no.6
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    • pp.839-844
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    • 2008
  • A strain SH-517 which produce extracellular keratinase, was isolated from the soil of a poultry waste and a poultry factory. An isolate SH-517 was identified as Bacillus sp. based on its morphological and biochemical characteristics. The optimal culture conditions for the production of keratinase by Bacillus sp. SH-517 were investigated. The optimal medium composition for keratinase production was determined to be 2.0% chicken feather as carbon source, 0.5% beef extract as organic nitrogen source, 0.5% $KNO_3$ as inorganic nitrogen source and 0.06% KCl, 0.05% NaCl, 0.04% $KH_2PO_4$, 0.03% $K_2HPO_4$ as mineral source and 0.01% yeast extract as growth factor. The optimal temperature and pH of medium were shown $40^{\circ}C$ and 8.5 with shaking culture (180 rpm/min), respectively. The maximum keratinase production reached maximum of 125 units/ml/min after 42 hr of cultivation under the optimal culturing conditions.