• Title/Summary/Keyword: Soil bacteria

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Improvement of Cyclosporin A Hydroxylation in Sebekia benihana by Conjugational Transfer of Streptomyces coelicolor SCO4967, a Secondary Metabolite Regulatory Gene (Sebekia benihana에서 Streptomyces coelicolor SCO4967 유전자 도입을 통한 하이드록실 사이클로스포린 A의 생전환)

  • Kim, Hyun-Bum;Lee, Mi-Jin;Han, Kyu-Boem;Kim, Eung-Soo
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.475-480
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    • 2010
  • Actinomycetes are Gram-positive soil bacteria and one of the most important industrial microorganisms due to superior biosynthetic capabilities of many valuable secondary metabolites as well as production of various valuable bioconversion enzymes. Among them are cytochrome P450 hydroxylase (CYP), which are hemoproteins encoded by a super family of genes, are universally distributed in most of the organisms from all biological kingdoms. Actinomycetes are a rich source of soluble CYP enzymes, which play critical roles in the bioactivation and detoxification of a wide variety of metabolite biosynthesis and xenobiotic transformation. Cyclosporin A (CyA), one of the most commonly-prescribed immunosuppressive drugs, was previously reported to be hydroxylated at the position of 4th N-methyl leucine by a rare actinomycetes called Sebekia benihana, leading to display different biological activity spectrum such as loss of immunosuppressive activities yet retaining hair growth-stimulating side effect. In order to improve this regio-selective CyA hydroxylation in S. benihana, previously-identified several secondary metabolite up-regulatory genes from Streptomyces coelicolor and S. avermitilis were heterologously overexpressed in S. benihana using an $ermE^*$ promoter-containing Streptomyces integrative expression vector. Among tested, SCO4967 encoding a conserved hypothetical protein significantly stimulated region-specific CyA hydroxylation in S. benihana, implying that some common regulatory systems functioning in both biosynthesis and bioconversion of secondary metabolite might be present in different actinomycetes species.

Rheinheimera aquatica sp. nov., Antimicrobial Activity-Producing Bacterium Isolated from Freshwater Culture Pond

  • Chen, Wen-Ming;Lin, Chang-Yi;Young, Chiu-Chung;Sheu, Shih-Yi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1386-1392
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    • 2010
  • A bacterial strain designated GR5$^T$, previously isolated from a freshwater culture pond in Taiwan while screening for bacteria for antimicrobial compounds, was characterized using a polyphasic taxonomic approach. Strain GR5$^T$ was found to be Gram-negative, aerobic, greenish-yellow colored, rod-shaped, and motile by means of a single polar flagellum. Growth occurred at $10-40^{\circ}C$ (optimum, $35^{\circ}C$), pH 7.0-8.0 (optimum pH 8.0), and with 0-2.0% NaCl (optimum, 0.5-1.0%). The major fatty acids were $C_{16:1}{\omega}7c$(36.3%), $C_{16:0}$(16.6%), $C_{12:0}$ 3-OH (12.5%), and $C_{18:1}{\omega}7c$(9.1%). The major respiratory quinone was Q-8, and the DNA G+C content of the genomic DNA was 51.9 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain GR5$^T$ belongs to the genus Rheinheimera, where its most closely related neighbors are Rheinheimera texasensis A62-14B$^T$ and Rheinheimera tangshanensis JA3-B52$^T$ with sequence similarities of 98.1% and 97.5%, respectively, and the sequence similarities to any other recognized species within Gammaproteobacteria are less than 96.5%. The mean level of DNA-DNA relatedness between strain GR5$^T$ and R. texasensis A62-14B$^T$, the strain most closely related to the isolate, was $26.5{\pm}7.6%$. Therefore, based on the phylogenetic and phenotypic data, strain GR5$^T$ should be classified as a novel species, for which the name Rheinheimera aquatica sp. nov. is proposed. The type strain is GR5$^T$ (=BCRC 80081$^T$=LMG 25379$^T$).

Utilization of Essential Oil Free Needles for Compost and Roughage (침엽정유추출잔사의 퇴비화 및 조사료 이용)

  • 최인규;강하영
    • Journal of Korea Foresty Energy
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    • v.21 no.1
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    • pp.7-15
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    • 2002
  • The essential oil free needles, which were left after distillation of essential oil from various coniferous needles, were fermented with food waste organics in order to use as compost and roughage. Microorganisms for the fermentation were selected from domestic sources such as swine compost, bark compost, and kimchi, etc, and consisted of aerothermophilic bacteria and actinomycetes. The weight reduction ratio of food waste organics treated with the microorganisms was 90% after 30 days treatment, and the fermentation temperature was kept at approximately $45^{\circ}C$. The compost process was really slow due to chemical compounds derived from needles, and it finally took 60 days for complete compost. When 10% of needle compost was mixed with soil for radish growth, the growth indicators such as leaf length and root weight were increased compared with control, while root weight, root width, and root length were inhibited on the addition of 20% needle compost. The nutrient value and digestibility ratio of various essential oil free needles as roughage for ruminant animals were evaluated. The ratio of crude protein for essential oil free needles from Korean pine(Pinus koraiensis) was 10.02%, which was higher than those of rice straw(5.48%) and corn(9.00%). The digestibility ratios of essential oil free needles from Sawara cypress(Chamaecwaris pislfera), Korean pine(Pinus koraiensis), and Japanese Red Pine(Pinus densiflora) was 53%, 34%, 34%, respectively, indicating that those essential oil free needles were considered as excellent roughage.

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Microbial Degradation of Polyethylene Glycol (Polyethylene Glycol의 미생물학적 분해)

  • 이종근;이상준;이재동;박송희;박재림
    • Korean Journal of Microbiology
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    • v.24 no.3
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    • pp.329-334
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    • 1986
  • The bacteria capable of utilizing polyethylene glycol(PEG) 6,000 as a sole carbon source were isolated from soil and sewage water connected to factory area. The isolate designated as EL-033 had high biodegradability on PEG 6,000, and was identified as Micrococcus sp. Micrococcus sp. EL-033 could grow on and degrade di-, tri-, tetraethylene glycols and PEGs with molecular weight up to 6,000 and very slowly stilize PEG 20,000 as sole carbon source, but not degrade ethylene glycol. The growth rate of isolate was increased in the higher molecular weight PEGs. The optical culture medium was established to be as follow: PEG 6,000, 0.2%(w/v); $K_2HPO_4$, 0.1%; $NaH_2PO_4{\cdot}12H_2O,\;0.1%\;:\;MgSO_4{\cdot}7H_2O$, 0.05%; polypeptone, 0.1% in distilled water, pH7.5. About 90% of PEG 6,000 was degraded in exponential phase of 48h culture and PEG 6,000 was completely degraded during 72h.

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An investigation of Panax ginseng Meyer growth promotion and the biocontrol potential of antagonistic bacteria against ginseng black spot

  • Sun, Zhuo;Yang, Limin;Zhang, Lianxue;Han, Mei
    • Journal of Ginseng Research
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    • v.42 no.3
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    • pp.304-311
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    • 2018
  • Background: Ginseng black spot disease resulting from Alternaria panax Whuetz is a common soil-borne disease, with an annual incidence rate higher than 20-30%. In this study, the bacterial strains with good antagonistic effect against A. panax are screened. Methods: A total of 285 bacterial strains isolated from ginseng rhizosphere soils were screened using the Kirby-Bauer disk diffusion method and the Oxford cup plate assay. We analyzed the antifungal spectrum of SZ-22 by confronting incubation. To evaluate the efficacy of biocontrol against ginseng black spot and for growth promotion by SZ-22, we performed pot experiments in a plastic greenhouse. Taxonomic position of SZ-22 was identified using morphology, physiological, and biochemical characteristics, 16S ribosomal DNA, and gyrB sequences. Results: SZ-22 (which was identified as Brevundimonas terrae) showed the strongest inhibition rate against A. panax, which showed 83.70% inhibition, and it also provided broad-spectrum antifungal effects. The inhibition efficacies of the SZ-22 bacterial suspension against ginseng black spot reached 82.47% inhibition, which is significantly higher than that of the 25% suspension concentrate azoxystrobin fungicide treatment (p < 0.05). Moreover, the SZ-22 bacterial suspension also caused ginseng plant growth promotion as well as root enhancement. Conclusion: Although the results of the outdoor pot-culture method were influenced by the pathogen inoculum density, the cropping history of the field site, and the weather conditions, B. terrae SZ-22 controlled ginseng black spot and promoted ginseng growth successfully. This study provides resource for the biocontrol of ginseng black spot.

Control of Ginseng Damping-off by Streptomyces sp. A3265 (방선균 A3265 균주에 의한 인삼 잘록병의 방제)

  • Woo, E-Eum;Lee, Gang-Seon;Lee, In-Kyoung;Choi, Jae-Eul;Yun, Bong-Sik
    • The Korean Journal of Mycology
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    • v.44 no.3
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    • pp.193-195
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    • 2016
  • Korean ginseng (Panax ginseng) possesses various biological and pharmacological properties. Damping-off is a critical disease on ginseng seedlings, which is caused by the fungal pathogens Rhizoctonia solani and Pythium sp.. This disease is generally controlled by the application of fungicides, but also biological control is an efficient and environmentally friendly way to prevent ginseng damping-off. In a previous study, we screened soil-borne bacteria with potential applications as biological control agents for ginseng damping-off and selected the bacterial strain Streptomyces sp. A3265, producing antifungal substances guanidylfungin and methylguanidylfungin. In this study, we investigated control efficacy of Streptomyces sp. A3265 against ginseng damping-off in the field. As a result, the incidence of damping-off was significantly reduced when soaking ginseng seeds in the culture broth of Streptomyces sp. A3265.

Studies on diaminododecane Utilization by Bacteria Studies on Diaminododecane Utilization by Corynebacterium sp. DAD 2-3 (Diaminododecane 자화균에 관한 연구 제2보 Corynebacterium sp. DAD 2-3의 Diaminododecane자화에 관한 연구)

  • 이상준;이종근
    • Korean Journal of Microbiology
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    • v.21 no.4
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    • pp.191-196
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    • 1983
  • A Corynebacterium sp. capable of utilizing diaminododecane (DAD) were isolated from the soil by enrichment culture. Among 9 different kinds of substituted alkanes containing CN, $NH_2$, Cl, and SH groups (monoterminally or diterminally substituted) tested as carbon source, the isolate, designated as DAD 2-3, utilized DAD, putrescine dihydrochloride, dodecane and laurylamine. Dodecanethiol, thioanisole, decanedithiol, dicyanooctane, laurylcyanide,and dichlorodecane were not utilized. When emulgen 950 was added to the medium, the growth of DAD 2-3 was slightly accelerated. Isolate DAD 2-3 grown in the medium with DAD as carbon source formed .alpha.-ketoglutaric acid. Metabolic product of DAD 2-3 grown in a medium without nitrogen source was different from that of grown in a medium with $NH_4NO_3$. When glucose, putrescine, n-dodecane and other alkane derivatives were tested in place of DAD, isolate DAD 2-3 yielded products different from those they formed with DAD suggesting specificity of DAD as a carbon source.

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Elimination of R-Plasmid in Streptomyces bobili (YS-40) by Ethldium Bromide (Ethidium Bromide에 의한 Streptomyces bobili(YS-40)의 R-Plasmid 제거)

  • 김상달;도재호
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.289-295
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    • 1982
  • Streptomyces bobili (YS-40) isolated from soil was tested that it had drug resistance against penicillin, cephalosporin series antibiotics and other antibiotics in the previous paper. The treatment of Streptomyces bobili, (YS-40) with ethidium bromide (EtBr), acriflavine and sodium dodecyl sulfate. (SDS) resulted in the elimination of R-plasmid from the host strain. Minimum growth inhibitory concentrations (MIC) of Hg, Ag, penicillin-G, ampicillin, chloramphenicol, oxytetracycline, streptomycin and kanamycin were found to be 15, 10, > 3, 000, > 100, > 1, 000, > 100, < 5 and < 5$\mu\textrm{g}$/$m\ell$ respectively. Among the curing agents, EtBr was proved to be the most powerful compound for the elimination of R-plasmid in the strain and the elimination rate with EtBr(10$\mu\textrm{g}$/$m\ell$) was about 98%. Optimal pH to. the elimination of R-plasmid was pH 7.0 and the R-plasmid in the cells incubated for 24 hrs was proved to be eliminated most effectively. Aerial mass color, soluble pigment formation and reverse side color were reported to be often the plasmid associated characteristics of the R-plasmid bearing bacteria. But these characteristics of the uncured and cured Streptomyces bobili, (YS-40) showed no changes in the most of the pigment formation media tested in this work.

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Purification and Characterization of the Antitumor Antibiotic from Streptomyces sp. YBE-316 (Streptomyces sp. YBE-316이 생산하는 항암성 항생물질의 정제 및 특성)

  • Park, Jae-Hong;Hahm, Byoung-Kwon;Bai, Dong-Hoon;Yu, Ju-Hyun
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.329-336
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    • 1995
  • For the development of new antitumor antibiotics produced by microorganisms, Streptomyces sp. YBE-316 was isolated from soil. The productivity of the antitumor antibiotic from Streptomyces sp. YBE-316 gradually increased after 60 hours, and was maximum after 100 hours after inoculation in growth medium (2.0% sucrose, 1.0% soybean meal, 0.1% K$_{2}$HPO$_{4}$, pH 7.0) at 30$\circ$C, 150 rpm, 5 NL/min by 30 l jar fermentor. This antitumor antibiotic was present only in mycelium, and stable in pH 5.0-10.0 for 20 minutes at 100$\circ$C. Antitumor and antibiotic activities were maintained at neutral pH, and heat stability was low. This antitumor antibiotic was soluble in methanol and ethanol, and insoluble in water, ethyl acetate, chloroform, and n-hexane. This antitumor antibiotic was sequentially purified by acetone extraction from mycelium, butanol extraction, and silica gel column chromatography. Antitumor activity was low against most tested cell lines, but antibiotic activity was high and low against yeasts and bacteria, respectivelv. The visualization test showed that this antitumor antibiotic had higher hydroxyl, ketone, amino, carboxyl groups, and sugar(s) in its structure. Instrumental analyses showed that this antitumor antibiotic was a pentaene in polyene class antibiotics. In pentaene class antibiotics, this was considered as an eurocidin or capacidin type antibiotics. The molecular weight of this antitumor antibiotic was higher than 683.0 daltons, and this antitumor antibiotic might be glycosylated by other sugar(s), instead of mycosamine or perosamine, an amino sugar.

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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.