• Title/Summary/Keyword: streptomyces

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Purification of Streptomyces Phospholipase D by Immunoaffinity Chromatoghraphy using Peptide Antibodies (Streptomyces phospholipase D의 정제를 위한 면역친화 크로마토그래피의 개발)

  • Park, In-Sun;Kim, Young-Ah;Jeong, Su-Jin;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.294-298
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    • 2006
  • An immunoaffinity chromatography for the specific binding of Streptomyces somaliensis phospholipase D (PLD) that is considered as an industrially potential enzyme was developed. By using the protein structure prediction programs and the X-ray crystal structure of a Streptomyces PLD, 5 different epitopes with high antigenicity that are predicted to locate on the surface of the S. somaliensis PLD were selected and then synthesized for the preparation of antipeptide antibodies. Each purified rabbit IgG was coupled with NHS-activated Sepharose to prepare the immunoaffinity resins. After one-step purification of the culture concentrate on the antipeptide IgG-coupled Sepharose column, SDS-PAGE and the Western blot analysis of the purified samples showed that purification of PLD on the affinity columns was satisfactory, indicating that the peptide design using the structural information of Streptomyces PLDs was rational. However, the purified PLD in the solution aggregated rapidly, which resulted in poor specific activity and low purification yield.

Isolation and Identification of Streptomyces sp. Producing Anti-vancomycin Resistant Staphylococcus aureus Substance (반코마이신 내성 Staphylococcus aureus 억제 물질 생산 Streptomyces sp.의 분리 및 동정)

  • Oh Se-Teak;Lee Jun-Jae;Lee Ji-Youn;Kim Jin-Kyu;Yang Si-Yong;Kim Yang-Soo;Song Min-Dong
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.90-95
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    • 2005
  • An Actinomycetes producing an anti-VRSA (vancomycin-resistant Staphylococcus aureus) substance was isolated from soil. The cultural, morphological, physiological and phylogenetic analyses of an isolated strain were investigated for identification. Cultural characteristics based on ISP (International Streptomyces Project) were as follows: white aerial mycelium, yellow reverse side, and good growth on various medium. Also, the isolate did not produce the soluble pigment. Morphological characteristics were showed cylindrical spore chain and smooth spore surface by SEM (Scanning Electron Microscope). Physiological characteristics were showed LL-type by DAP isomer analysis and detected glycine, glutamic acid and alanine. A phylogenetic analysis of the 16S rDNA provided a clue that the isolated strain was actually a member of the genus Streptomyces, because the determined sequence exhibited a higher homology with Streptomyces echinatus. The isolate was identified to be a genus of Streptomyces sp.. The optimal culture conditions for the maximum production of anti-VRSA substance by Streptomyces sp. were attained in a culture medium composed of $2.0\%$ (w/v) glucose, and $0.4\%$ (w/v) yeast extract. The anti-VRSA substance was highly produced after 5 days of culture. Optimal pH and temperature conditions for the production of anti-VRSA substance were pH 7.0 and $28^{\circ}C$, respectively.

In vivo Functional Analysis of γ-butyrolactone Autoregulator Receptor Gene (scaR) in Streptomyces clavuligerus (Streptomyces clavuligerus의 γ-butyrolactone autoregulator receptor 유전자에 대한 in vivo 기능 분석)

  • Kang Su-Jin;Lee Chang-Kwon;Choi Sun-Uk;Kim Hyun-Soo;Hwang Yong-Il
    • Journal of Life Science
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    • v.16 no.1
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    • pp.76-81
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    • 2006
  • A $\gamma-butyrolactone$ autoregulator receptor has a common activity as DNA-binding transcriptional repressors controlling secondary metabolism and/or morphological differentiation in Streptomyces. A gene (scaR) encoding it was cloned from Streptomyces cravuligerus, a clavulanic acid producer, and was in vitro characterized in a previous report. In this study to clarify the in vivo function of ScaR, a $\gamma-butyrolactone$ autoregulator receptor of Streptomyces clavuligerus, we constructed a scaR-deleted strain by means of homologous recombination. No difference in morphology was found between the wild-type strain and the scaR-disruptant, but the scaR-disruptant showed higher clavulanic acid production. This indicates that the ScaR in S. clavuligerus acts as a negative regulator of the biosynthesis of clavulanic acid, but plays no role in morphological differentiation.

Development of Doxorubicin overproducing Streptomyces Strain using Protoplast Regeneration (방선균 원형질체 재생에 의한 독소루비신 고생산성 균주개발)

  • 박희섭;박현주;김용훈;임상민;김동일;류욱상;김상린;김응수
    • KSBB Journal
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    • v.18 no.4
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    • pp.289-293
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    • 2003
  • Doxorubicin is an anthracycline-family polyketide compound with a very potent anti-cancer activity, typically produced by Streptomyces peucetius. In order to increase doxurubicin productivity, a semi-industrial doxorubicin-producing Streptomyces strain named BR-Dox was cultured in a R2YE liquid medium containing CaCO$_3$, and then converted to a cell wall-free protoplast using lysozyme treatment method, followed by PEG-mediated cell wall regeneration. Among several protoplast-regenerated Streptomyces BR-Dox strains, two independent isolates named BR-Dox4 and BR-Dox6 were visually selected using thin layer chromatography (TLC) based on the pigment overproducing phenotype. Comparing with Streptomyces BR-Dox parental strain, two protoplast-regenerated strains, BR-Dox4 and BR-Dox6 exhibited 25.2% and 12.2% higher doxorubicin productivity analyzed by high pressure liquid chromatography (HPLC), respectively. This result suggests that a protoplast-regeneration of an antibiotics-producing Streptomyces strain should be a promising strain development approach for antibiotics overproduction in Streptomyces species.

Studies on the Classification and Identification of Actinomycetes Producing Chitinase isolated from Soil (토양(土壞)에서 분리(分離)한 Chitinase 생성(生成) 방선균(放線菌)의 분류동정(分類同定))

  • Kim, Yeong-Yil;Kim, Yong-Woong;Kim, Kwang-Sick
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.3
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    • pp.339-345
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    • 1988
  • Four actinomycetes which produces extracellular chitinase were isolated from soil and organic matter all over the Chonnam provincial area. The chemical composition, morphological, cultural and physiological properties of isolated strain S-25, S-42, S-172 and S-267 were studied in relation to the toxonomical properties. All of strains contained phospholipids such as phosphatidyl ethanolamine, phosphatidyl inositol and diphosphatidyl glycerol. The components of cell wall in all strains have L, L-DAP, Glutamic acid, Alanine, Glycine and Glucosamine. The surface of spore is smooth and colony is grey in all strains, Based on the results obtained in these experiments all of strains are identified as Streptomyces sp.S-25, Streptomyces sp.S-42, Streptomyces sp.S-l72 and Streptomyces sp. S-267.

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Antibacterial Activity of Streptomyces Strains Isolated from Different Regions of Jordan

  • Hala Khyami-Horani;Amal Al-Aboudi;Musa Abu Zarga;Monther Sadder;Halima Othman
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.474-483
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    • 2023
  • Members of the genus Streptomyces produce more than 70% of antibiotics. The rise in antibiotic resistance globally enhanced the search for novel species with the ability to produce new bioactive compounds. This study was initiated to investigate different regions in Jordan for previously uncultured and rare Streptomyces species capable of producing novel antimicrobial compounds especially active against bacteria resistant to antibiotics. A total of 191 Streptomyces strains were isolated from 26 soil samples collected from different geographic regions in Jordan. Isolates were characterized based on colony and cellular morphology as well as using 16S rRNA gene sequencing. These isolates were screened for their ability to produce antibiotics by the perpendicular-cross streak method, and then tested by well diffusion method against tested pathogens. Fifty-four isolates showed potential to produce antimicrobial products especially active against resistant bacteria, 20.1% of the isolates showed inhibitory effect against Staphylococcus aureus, 16.9% against clinical MSSA strains, and 18.0% against MRSA: whereas only 4.2% against Esherichia coli, 3.2% against Klebsiella pneumonia, 2.7% against Pseudomonas aeruginosa, and 10.0% against clinical Candida albicans. Three isolates were selected for further identification due to their antibacterial activity against S. aureus, MRSA, and MSSA. These isolates were identified as follows; Streptomyces aburaviensis DSa3, Streptomyces alboniger SAb7 and Streptomyces misionensis ZAb2, based on cultural, biochemical characteristics and molecular analysis of the 16S rRNA.

Nucleotide Sequence of 16S rRNA Gene from Streptomyces melanosporofaciens 7489

  • LEE, DONG-SUN;SUNG-OUI SUH;SEON-KAP HWANG;TAEG-KYU KWON;TAE-HO KIM;WOO-CHANG SHIN;SOON-DUCK HONG
    • Journal of Microbiology and Biotechnology
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    • v.6 no.5
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    • pp.364-365
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    • 1996
  • A region encoding the 16S rRNA was cloned by PCR from Streptomyces melanosporofaciens 7489 and sequenced by the chain-termination dideoxy sequencing method. A phylogenetic tree constructed by sequence alignment of 24 Streptomyces species suggests that there is little evolutionary distance between this strain and Streptomyces rimosus.

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Selection and Identification of a Strain KT-10 Producing the Cathepsin B Inhibitor

  • Han, Kil-Hwan;Do, Jae-Ho;Kim, Sang-Dal
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.333-340
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    • 1997
  • An actinomycetes, KT-10 isolated from ginseng field in Kyongpook, Korea was selected based on its ability to produce a lysosomal cathepsin B inhibitor. The inhibitor purified from the culture supernatant of the isolate KT-10 showed strong inhibitory effects against cathepsin B as well as against papain when the activities were measured using synthetic substrate, ${\alpha}$-N-benzyloxycarbonyl-L-Iysine p-nitrophenyl ester (CLN) or ${\alpha}$-N-benzoyl-D,L-arginine 2-naphthylamide (BANA). The isolate KT-10 was identified as a species of Streptomyces based on its morphological characteristics and chemotaxonomic data. The TAXON program of Ward was used to identify Streptomyces sp. KT-10 as a strain of Streptomyces luteogriseus belong to cluster 18 of the genus Streptomyces with a Willcox probability 0.999388. The cathepsin B inhibitor was presumed to a novel material composed of a polyhydroxylamine.

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