• Title/Summary/Keyword: screening of strain

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Isolation and Properties of Amino Acid Antimetabolite from Streptomyces sp. YS-943 (Streptomyces sp. YS-943균주가 생산하는 아미노산 대사길항물질의 정제와 성상)

  • 유성재;박부길
    • Microbiology and Biotechnology Letters
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    • v.23 no.1
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    • pp.81-86
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    • 1995
  • A Streptomyces strain YS-943, which produced amino acid antimetabolite, was isolated from soil. During the course of screening for new amino acid antimetabolites from the culture broth of Actinomycetes, we found that the strain produced a substance active against Gram-positive bacteria and its activity was reversed by L-methionine and L-histidine on the synthetic minimal agar medium in the culture broth.The morphological and cultural characteristics serve to identify the producing organism strain YS-943 as the genus Streptomyces. Fermentation was carried out in the synthetic medium at 28$\CIRC$C for 48 hours. The fermentation yield reached about 12 mg per liter of the broth. The YS-943 substance was obtained as white powder, mp 194$\CIRC$C and has the molecular formular of C$_{4}$H$_{8}$N$_{2}$O$_{4}$. Its structure was determined to be o-carbamyl-D-serine by spectroscopic data. It is active against some Gram-positive bacteria and reversed by L-methionine and L-histidine.

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Studies on antibiotics against Wheat black rust (I) (밀의 항흑수병 항생물질의 연구 1)

  • 정영기
    • Korean Journal of Microbiology
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    • v.19 no.3
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    • pp.108-114
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    • 1981
  • In order to isolate microorganisms which produce antibiotics aganist wheat black rust, some bacteria, molds, and actinomycetes were isolated from soils and screened for the production of antibiptics against wheat black rust. Beacuse wheat black rust-puccinia graminis--is a complete parsitic mold which can't grow in artifical medium, new method for the screening of antibiotic producing microorgsnisms against wheat black rust developed by using live leaves of wheat. With new method, a strain No. $480HS_{20}$ which produces a substnace having strong and Puccinia graminis activity and very narrow antimicrobial spectrum was isolated. the substance produced by the strain No.$480HS_{20}$ had better anti Puccinia graminis activity than any other known antifungal antibotics such as kasurgamycin, balasticidins, actidione, antimycin, ologomycin. And the substance was observed to be very stable at heat and ultraviolet light. The strain was indentified as Bacillus subtilis.

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Genome Shuffling of Mangrove Endophytic Aspergillus luchuensis MERV10 for Improving the Cholesterol-Lowering Agent Lovastatin under Solid State Fermentation

  • El-Gendy, Mervat Morsy Abbas Ahmed;Al-Zahrani, Hind A.A.;El-Bondkly, Ahmed Mohamed Ahmed
    • Mycobiology
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    • v.44 no.3
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    • pp.171-179
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    • 2016
  • In the screening of marine mangrove derived fungi for lovastatin productivity, endophytic Aspergillus luchuensis MERV10 exhibited the highest lovastatin productivity (9.5 mg/gds) in solid state fermentation (SSF) using rice bran. Aspergillus luchuensis MERV10 was used as the parental strain in which to induce genetic variabilities after application of different mixtures as well as doses of mutagens followed by three successive rounds of genome shuffling. Four potent mutants, UN6, UN28, NE11, and NE23, with lovastatin productivity equal to 2.0-, 2.11-, 1.95-, and 2.11-fold higher than the parental strain, respectively, were applied for three rounds of genome shuffling as the initial mutants. Four hereditarily stable recombinants (F3/3, F3/7, F3/9, and F3/13) were obtained with lovastatin productivity equal to 50.8, 57.0, 49.7, and 51.0 mg/gds, respectively. Recombinant strain F3/7 yielded 57.0 mg/gds of lovastatin, which is 6-fold and 2.85-fold higher, respectively, than the initial parental strain and the highest mutants UN28 and NE23. It was therefore selected for the optimization of lovastatin production through improvement of SSF parameters. Lovastatin productivity was increased 32-fold through strain improvement methods, including mutations and three successive rounds of genome shuffling followed by optimizing SSF factors.

Isolation and Numerical Identification of Streptomyces humidus strain S5-55 Antagonistic to Plant Pathogenic Fungi

  • Lim, Song-Won;Kim, Jeong-Dong;Kim, Biom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.16 no.4
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    • pp.189-199
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    • 2000
  • To search for the antifungal substances, various actino-mycete isolates were obtained from various soils of Korea using plate dilution method on the humic acid vitamin agar plates. In the screening procedures using a dual culture method, 32 actionomycete isolates were selected, which showed the inhibitory activity against mycelial growth of plant pathogenic fungi Altirnaria mali, Colletotrichum gloeosporides, Fusarium oxysporum f.sp. cucumerinum, Magnaporthe grisea, Phytophthora capsici, and Rhizoctonia solani. Bioassay of the crude extracts from culture filtrates and mycelial mets revealed that 12 antagonistic actionomycetes produced highly active antifungal substances. Actinomycete strain S5-55 which showed the substantial antifungal activity against the tested fungi was selected for production of the antifungal substances. Based on the cytochemical and morphological characteristics, strain S5-55 was identified as a Streptomyces species. The results of the numerical identification using the TAXON program confirmed that Streptomyces strain S5-55 was identical with Streptomyces humidus including in TAXON major cluster 19. The production of antifungal substance was most favorable when S. humidus strain S5-55 was cultivated for 10 dats on soluble starch broth supplemented with $K_2$HPO$_4$. The antifungal substances active against the plant pathogenic fungi P. capsici and M. grisea were partially purified using $\textrm{C}_{18}$ reversed-phase column chromatography.

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Effective Screening of Antagonist for the Biological Control of Soilborne Infectious Disease (Damping-Off)

  • LEE BAEK-SEOK;LEE HYANG-BOK;CHOI SUNG-WON;YUN HYUN-SHIK;KIM EUN-KI
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.701-709
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    • 2005
  • An efficient method of selecting an antagonistic strain for use as a biological control agent strain was developed. In this improved method, the surface tension reduction potential of an isolate was included in the 'decision factor,' in addition to two other factors; the growth rate and pathogen inhibition. By using a statistically designed method, an isolate from the soil was selected and identified as Bacillus sp. GB 16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth were observed when the Bacillus sp. GB 16 was used. The action of the surface tension reducing component was assumed to enhance the wetting, spreading, and residing of the antagonistic strain in the rhizosphere. This result showed that the improved selection method was quite effective in selecting the best antagonistic strain for the biological control of soilborne infectious plant pathogens.

Isolation of Novel Pseudomonas diminuta KAC-1 Strain Producing Glutaryl 7-Aminocephalosporanic Acid Acylase

  • Kim, Dae-Weon;Kang, Sang-Mo;Yoon, Ki-Hong
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.200-205
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    • 1999
  • 7-Aminocephalosporanic acid (7-ACA) is the initial compound in preparation of cephalosporin antibiotics widely used in clinical treatment. Bacteria producing glutaryl 7-ACA acylase, which convert cephalosporin C to 7-ACA, has been screened in soil samples. A bacterial strain exhibiting high glutaryl 7-ACA acylase activity, designated KAC-1, was isolated and identified as a strain of Pseudomonas diminuta by characterizing its morphological and physiological properties. The screening procedures include culturing on enrichment media containing glutaric acid, glutamate, and glutaryl 7-aminocephalosporanic acid as selective carbon sources. To enhance enzyme production, optimal cultivation conditions were investigated. This strain grew optimally at pH 7 to 9 and in temperatures of 20 to 40 C, but acylase production was higher when the strain was grown at 25 C. Glutaric acid, glutamate and glucos also acted as inducers for acylase production. In a jar fermenter culture, P. diminuta KAC-1 produce acylase in a growth-associated manner. The substrate specificity of KAC-1 acylase by cell extract showed that this enzyme had specificity toward glutaryl 7-ACA, glutaryl 7-ADCA, but not cephalosporin C.

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Screening and Characteristics of a Mutant of Actinoplanes teichomyceticus ATCC31121 Highly Producing Teicoplanin (Teicoplanin 생산성이 우수한 Actinoplanes teichomyceticus ATCC31121 변이주 선별 및 배양학적 특성)

  • 노용택
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.299-304
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    • 2001
  • Teicoplanin is a kind of glycopeptide antibiotics produced by Actinoplanes teichomyceticus, and used in the clinical antibiotic such as vancomycin against methicillin-resistant Stabphylococcus aureus (MRSA). Actino planes teichomyceticus ATCC 31121 was mutated with UV to obtain a superior mutant strain with increased level of teicoplanin production. In this investigation, lethal curve was obtained and the optimal condition to induce mutagenesis was determined to isolate the desirable mutant strain. It was also confirmed that teicoplanin activities by agar diffusion method was compared with the parent strain. One mutant strain, T991014-1 with the highest productivity, was finally selected, and was characterized through the various tests such as amylase activity, protease activity, halotolerance, antibiotic resistance, autotoxicity, and productivity. Ad fermentation characteristics of the mutant strain were also studied.

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Concanamycin B, Active substance Against Phytophthora capsici Produced by Streptomyces neyagawaensis 38D10 Strain (Streptomyces neyagawaensis 38D10 균주가 생산하는 concanamycin B의 항고추역병 활성)

  • Kim, Chang-Jin;Lee, In-Kyoung;Yun, Bong-Sik;Yoo, Ick-Dong
    • Microbiology and Biotechnology Letters
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    • v.21 no.4
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    • pp.322-328
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    • 1993
  • During the screening of antifungal compounds from microbial secondary metabolites to control phytophthora blight of red pepper caused by Phytophthora capsici, a soil isolate, strain 38D10 was selected. Based on taxonomic studies, this strain was identified as Streptomyces neyagawaensis. The antifungal compound was purified from culture broth by HP-20 column chromatography, ethyl acetate extraction, silica gel column chromatography, HPLC and identified as concanamycin B by UV. $^1H$-NMR, $^{13}C$-NMR, SIMS analysis. Concanamycin B has strong antifungal activity against some phytopathogenic fungi but not antivacterial activity and preventive value were 50% and 100% at 125ppm and 250ppm in pot assay.

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Use of Bioluminescent Indicator Acinetobacter Bacterium for Screening and Characterization of Active Antimicrobial Agents

  • Haleem Abd-El;A.M. Desouky;Zaki Sahar A.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1706-1712
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    • 2006
  • Because of the need for new antimicrobial substances with novel mechanisms of action, we report here the use of an Acinetobacter reporter system for high-throughput screening of active antimicrobial agents. The bioreporter Acinetobacter strain DF4/PUTK2 carrying luciferase genes luxCDABE was chosen because of its ecological importance and it is widespread in nature. This bioreporter is genetically engineered to emit light constitutively that can be measured in real time by luminometry. Hence, this reporter system was employed to determine the bacteriostatic actions of spent-culture supernatants derived from twelve bacterial isolates. Out of the results, the strongest bioluminescence inhibitory effect of the supernatants was recorded with Bacillus cereus strain BAC (S5). Subsequently, ethyl acetate extracts of extracellular products of strain BAC (S5) were separated by a thin-layer chromatography (TLC). Based on the bioluminescence inhibitory assay, three fractions were found to have antimicrobial activity. One fraction (C) having the strongest antimicrobial activity was further purified using TLC and characterized by IR, $^1H$ NMR, mass spectrometry, SDS-PAGE, and amino acid composition analysis. The results predicted the presence of 2-pyrrolidone-S-carboxylic acid (PCA) and the octadeconic-acid-like fatty acid. Fraction C also demonstrated a broad inhibitory activity on several Gram-negative and Gram-positive bacteria. In conclusion, the Acinetobacter reporter system shows great potential to be a reliable, sensitive, and real-time indicator of the bacteriostatic actions of the antimicrobial agents.

Isolation of Methylotrophic Actionmycetes Capable of Producing Antagonistic Activity Against Oral Resident Bacteria and Screening of Mutants (구강상재균을 중심으로 항균력을 나타내는 메탄올 자화방선균의 분리 및 변이주 생산)

  • Park, Myoung-Ho;Lee, Hwa-Sik;Bae, Bong-Jin;Kim, Joung
    • Journal of Technologic Dentistry
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    • v.22 no.1
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    • pp.145-152
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    • 2000
  • In order to select an effective antibiotic substance against oral resident bacteria, we were isolated from soil and texonomically analyzed. Seven hundred and eighteen strains were isolated on humic acid- vitamin agar(HV agar) and 220 strains were on methanol medium from three each paddy forest, field and riverside soil samples. So, during the screening of antibiotics from soil, we isolated microorganisms showing powerful antagonistic activity against oral resident bacteria. Microorganism was tested against 25 strains of bacteria, yeast and fungi. Among them, No. 248 strain exhibited the most strongly growth inhibition. So, the taxonomical analysis the isolated strain was found to be unknown Actinomyces sp. and was named No 248. A production of the antibiotics from No. 248 begins at the early exponential phase developed at the 72th hour under the optinum conditions. The property of No. 248 antimicrobial compound was very stable under acid(pH 3.0) and alkali(pH 10.0) treatment, but it was instable in heat treatment at $120^{\circ}C$. For the improvement of antibiotic activity, two mutants were isolated from strain No. 248 by the treatment of mutagenic agents, NTG and hydroxylamine. As a result, the mutant strains excreted the potent antibiotics to inhibit the growth of Candida albicans.

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