• Title/Summary/Keyword: spore production

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Isolation and Culture Conditions of Hydrogen Producing Bacterium Enterobacter sp. ES392 (수소생산균 Enterobacter sp. ES392의 분리 및 배양조건)

  • Jeon, Sung-Jong;Lee, Eon-Seok
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.399-404
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    • 2010
  • A hydrogen-producing bacterium (strain ES392) was isolated from pond water located in the Dong-Eui University, Busan, Korea. The cell was long-rod type ($1.4\;{\mu}m$) of about ($0.6\;{\mu}m$) in diameter, and not formed flagellum and spore. Phylogenetic analysis based on the 16S rRNA sequence and biochemical studies indicated that ES392 belonged to the genus Enterobacter sp. The optimum pH and temperature for hydrogen production was 7.5 and $35^{\circ}C$, respectively. The optimization of medium compositions which maximize hydrogen production from Enterobacter sp. ES392 was determined. As a result, the maximum hydrogen production was obtained under the conditions of 4% (w/v) sucrose, 0.5% (w/v) yeast extract and 50 mM potassium phosphate buffer (pH 7.5). Under batch culture conditions, the maximal hydrogen production and yield were obtained as 3481 mL/L and 1.33 mol/mol sucrose, respectively.

Monoclonal Antibody-Based Indirect-ELISA for Early Detection, Diagnosis and Monitoring of Epiphytic Didymella bryoniae in Cucurbits.

  • Lee, Seon-Chul;Shim, Chang-Ki;Kim, Dong-Kil;Bae, Dong-Won;Kyo, Seo-Il;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.133.1-133
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    • 2003
  • Gummy stem blight, caused by Didymella bryoniae occurs exclusively on cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. Through this study, we optimized cultural conditions for mass-production of pycnidiospore by Metal Halide Lamp irradiation. In brief, the mycelial was cultured at $26^{\circ}C$ on PDA, for 2 days under the darkness, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$, a great number of pycnidia was simultaneously formed. Thus produced pycnidiospores were used as immunogen. From fusions of myeloma cell (v-653) with splenocytes from immunifed mice were car ried out. And, two hybridoma cell lines that recognized the immunogen Didymella bryoniae were obtained. One Monoclonal Antibody, Db1, recognized the supernatant and the other monoclonal antibody, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid two MAb Db1 and Db15, were immunotyped and identified as IgG1 and IgG2b, respectively. Titer of MAb Db1 and MAb Db15 was measured absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with Didymella bryoniae but none reacted with other of fungi and CMV, CGMMV Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{3}$ well by indirect ELISA characterization of the MAb Db1, Db15 antigen by heat and protease treatments show that the epitope recognized by the MAb Bb1, Db15 were a glycoprotein.

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Effect of crude ginseng saponin and raw ginseng juice on the growth of ginseng root rot organisms, Fusarium solani and Erwinia carotovora (인삼 조 Saponin과 조즙액이 인삼근부병균 Fusarium solani와 Erwinia carotovora의 생육에 미치는 영향)

  • Park Chang-Seuk;Ohh Seung-Hwan
    • Korean journal of applied entomology
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    • v.20 no.1 s.46
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    • pp.1-5
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    • 1981
  • Effect of crude saponin and raw ginseng juice on root rot pathogens such as Fusarium solani and Erwinia carotovora for there growth or spore germination was investigated. Macroconidial germination of F. solani was decreased as the incrasee of the crude saponin concentration. especially, percentage of the germination was remarkably reduced when the concentration was more than 500ppm. The spore production of F. solani was also reduced as the increase of the crude saponin concentration and this phenomenon was particularly profound on a solid medium. Mycelial growth was decreased when the crude saponin was added, while the effect of the concentration was not apparently significant. The higher concentration of raw ginseng juice is, the more the growth of F. solani. Growth of E. carotovora was enhanced by crude ginseng saponin and raw ginseng juice. The crude ginseng saponin stimulated the growth of E. carotovora as the increase of the concentration, while more growth of the bacteria obtained at $1\%$ raw ginseng juice added.

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Effect of water potential on mycelial growth, reproduction, and spore germination by Cylindrocladium crotalariae (Cylindrocladium crotalariae의 균사자람, 포자형성과 포자발아에 대한 Water potential의 효과)

  • Sung Jae Mo;Heo No Youl;Kim Se Keun
    • Korean journal of applied entomology
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    • v.20 no.1 s.46
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    • pp.37-41
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    • 1981
  • Mycelial growth and production of macroconidia by Cylindrocladium crotalariae on either PDA or water agar containing soybean leaf pieces adjusted to different water potentials with KCI. were consistently maximal at -14bars. Mycelial growth by this fungus was nii at about -100 bars and below at 30C, -80 bars and below at 25C and -64 bars and below at 20C. Sporulation was prevented at -64 bars and below at $20^{\circ}C\;and\;25^{\circ}C$. Perithecial andmicrosclerotial formation was maximal at abot -1.4 to - 3.0 bars (the basal medium without sait). Percentage of spore germination for this fungus was uniformly maximal at all water potentials between -1.4 bars (the highest tested) and - 20 bars, was progressively less as the water potential lowered below -20 bars, and was prevented at - 60 bars.

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Monoclonal Antibody-Based Indirect-ELISA for Early Detection and Diagnosis of Epiphytic Didymella bryoniae in Cucurbits

  • Lee, Sun-Cheol;Han, Ki-Soo;Lee, Jung-Han;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.19 no.5
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    • pp.260-265
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    • 2003
  • Gummy stem blight caused by Didymella bryoniae occurs exclusively in cucurbits. This fungus has been known not to produce its pycnidium in vitro unless irradiated. In this study, cultural conditions for the mass-production of pycnidiospore by Metal Halide (MH) lamp irradiation were maximized. The mycelia were cultured at $26^{\circ}C$ on PDA for 2 days under dark condition, and then the plate was illuminated with MH lamp continuously for 3-4 days at $26^{\circ}C$. Results show that a great number of pycnidia were simultaneously formed. The pycnidiospores produced were then used as immunogen. Fusions of myeloma cell (v-653) with splenocytes from immunized mice were carried out. Two hybridoma cell lines that recognized the immunogen D. bryoniae were obtained. One monoclonal antibody (MAb), Dbl, recognized the supernatant while another MAb, Db15, recognized the spore. Two clones were selected which were used to produce ascite fluid of the two MAb, Dbl and Db15, the immunotypes of which were identified as IgG1 and IgG2b, respectively. Titers of MAb Dbl and MAb Db15 were measured and the absorbance exceeded 0.5 even at a $10^{-5}$ dilution. The MAbs reacted positively with D. bryoniae but none reacted with other viral isolates, Cucumber mosaic virus and Cucumber green mottle mosaic virus. Sensitivity of MAb was precise enough to detect spore concentration as low as $10^{-3}$/well by indirect ELISA. Characterization of the MAbs Dbl, Db15 antigen by heat and protease treatments, which suggests that the epitope recognized by these two MAbs was glycoprotein.

Improvement of sporeless strain in oyster mushroom Pleurotus ostreatus (느타리버섯 소포자 형성 균주 개발)

  • Shin, Pyung-Gyun;Oh, Se-Jong;Yoo, Young-Bok
    • Journal of Mushroom
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    • v.4 no.2
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    • pp.53-56
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    • 2006
  • The enormous production of spores by the fruitbodies in the cultivation of oyster mushrooms (Pleurotus ostreatus) is develop an allergy with symptoms similar to an "extrinsic allergic alveolitis (EEA)". the sporeless strain would noy only benefit health of mushroom workers but also reduce the risk of viral infections on the mushroom farms. For the development of a sporeless strain of P. ostreatus we used strain ASI 2069. This non-commercial strain is completely nonsporulating. We have recovered both nuclear types of strain ASI 2069 as monokaryons (hereafter referred to as neohaplonts) by protoplasting the mycelium. Crosses between neohaplonts and SSI's(single spore isolates) obtained from a sporulating commercial strain ASI 2180 yielded fruitbodies that isolated 128 strains. 13 excellent strains are selected from 30 bred strains by quality of fruitbodies and spore number. Among 13 excellent strains, G192 strain is chose finally to high yield and sporelessness.

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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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Effect of Tenderizer on Physical Quality and Microbial Safety during Korean Beef Jerky Production (한국형 우육포의 제조공정 중 연화제가 육포품질 및 미생물학적 안전성에 미치는 영향)

  • Kim, Hyoun-Wook;Han, Doo-Joung;Kim, Cheon-Jei;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.28 no.5
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    • pp.675-680
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    • 2008
  • The physical quality and microbial safety of Korean beef jerky was evaluated at various steps during its preparation. Microbial counts in raw beef demonstrated mesophillic bacteria at 4.20 Log CFU/g, psychrotrophic bacteria at 3.85 Log CFU/g, anaerobic bacteria at 4.90 Log CFU/g, and yeast and molds at 1.92 Log CFU/g. Spore-forming bacteria and coliforms were not detected in raw beef samples. Spices and spiced meats showed similar trends in microbial counts, demonstrating minimal microbial contamination during these stages of preparation. The final beef jerky product exhibited counts of mesophillic bacteria at 1.15-1.66 Log CFU/g, psychrotrophic bacteria at 1.15-1.66 Log CFU/g, and anaerobic bacteria at 0.81-1.72 Log CFU/g. Spore-forming bacteria, yeast and molds, and coliforms were not detected in beef jerky. Significant differences from added ingredients occurred for instron textural profile analysis traits for hardness. In general, Korean beef jerky with humectant and tenderizer had lower hardness than control (without humectant and tenderizer). Also, the sample added with 0.01% protease from Streptomyces griseus had lower hardness than all samples. All samples had 0.7l to 0.72 water activities, and the color and pH were not shown in significant changes of all samples.

Inhibition of Bacillus cereus Growth and Toxin Production by Bacillus amyloliquefaciens RD7-7 in Fermented Soybean Products

  • Eom, Jeong Seon;Choi, Hye Sun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.1
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    • pp.44-55
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    • 2016
  • Bacillus cereus is a gram-positive, rod-shaped, spore-forming bacterium that has been isolated from contaminated fermented soybean food products and from the environment. B. cereus produces diarrheal and emetic toxins and has caused many outbreaks of foodborne diseases. In this study, we investigated whether B. amyloliquefaciens RD7-7, isolated from rice doenjang (Korean fermented soybean paste), a traditional Korean fermented soybean food, shows antimicrobial activity against B. cereus and regulates its toxin gene expression. B. amyloliquefaciens RD7-7 exhibited strong antibacterial activity against B. cereus and inhibited the expression of B. cereus toxin-related genes (groEL, nheA, nheC, and entFM). We also found that addition of water extracts of soybean and buckwheat soksungjang (Korean fermented soybean paste made in a short time) fermented with B. amyloliquefaciens RD7-7 significantly reduced the growth and toxin expression of B. cereus. These results indicate that B. amyloliquefaciens RD7-7 could be used to control B. cereus growth and toxin production in the fermented soybean food industry. Our findings also provide a basis for the development of candidate biological control agents against B. cereus to improve the safety of fermented soybean food products.

Identification and Production of Constitutive Chitosanase from Bacillus sp. HW-002

  • Lee, Hyean Woo;Jong Whan Choi;Dong Pyou Han;Noo Woon Lee;Sung Lim Park;Dong Heui Yi
    • Journal of Microbiology and Biotechnology
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    • v.6 no.1
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    • pp.12-18
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    • 1996
  • A chitosanase-producing bacteria was isolated on chitosan agar plate from soil samples. The strain was spore-forming gram positive bacteria, catalase positive, and rod shape. The strain was identified as Bacillus cereus. The strain did not need an inducer for the synthesis of chitosanase. Chitosanase from Bacillus sp. HW-002 was constitutive enzyme. The optimal medium for the production of the enzyme was composed of 0.5$\%$ sucrose and $1.5\%$ yeast extract-tryptone (1:1 w/w) mixture at pH 6.5. After Bacillus sp. HW-002 was cultivated at $32^{\circ}C$ for 32 h, maximal productivity was gained to be about 27, 200 U/l. Chitosanase from Bacillus sp. HW-002 was a mixed growth-linked metabolite.

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