• 제목/요약/키워드: Streptomyces venezuelae

검색결과 20건 처리시간 0.028초

Biosynthesis of Plant-Specific Flavones and Flavonols in Streptomyces venezuelae

  • Park, Sung-Ryeol;Paik, Ji-Hye;Ahn, Mi-Sun;Park, Je-Won;Yoon, Yeo-Joon
    • Journal of Microbiology and Biotechnology
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    • 제20권9호
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    • pp.1295-1299
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    • 2010
  • Recently, recombinant Streptomyces venezuelae has been established as a heterologous host for microbial production of flavanones and stilbenes, a class of plant-specific polyketides. In the present work, we expanded the applicability of the S. venezuelae system to the production of more diverse plant polyketides including flavones and flavonols. A plasmid with the synthetic codon-optimized flavone synthase I gene from Petroselium crispum was introduced to S. venezuelae DHS2001 bearing a deletion of the native pikromycin polyketide synthase gene, and the resulting strain generated flavones from exogenously fed flavanones. In addition, a recombinant S. venezuelae mutant expressing a codon-optimized flavanone $3{\beta}$-hydroxylase gene from Citrus siensis and a flavonol synthase gene from Citrus unshius also successfully produced flavonols.

Streptomyces Peucetius에서의 ${\varepsilon}$-rhodomycinone 추출 및 이종균주에서의 rhodomycin D 생산 연구 (NDP-sugar production and glycosylation of ${\varepsilon}$-rhodomycinone in Streptomyces venezuelae)

  • 박성희;차민호;김은정;윤여준;송재경;이희찬;류광경;김병기
    • KSBB Journal
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    • 제23권1호
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    • pp.44-47
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    • 2008
  • Streptomyces peucetius가 생산하는 anthracycline 계열의 doxorubicin은 치료목적으로 사용되는 중요한 항암제 중 하나이다. Doxorubicin은 rhodomycin D에서부터 몇 단계의 생합성 과정을 더 거쳐 생산되는데, 생물학적 활성을 갖기 위해서는 deoxy-sugar의 전이가 반드시 일어나야 한다. 본 논문에서는 이종균주인 Streptomyces venezuelae에 11개의 유전자를 형질 전환하여 TDP-L-daunosamine를 생산하고 이것을 ${\varepsilon}$-rhodomycinone에 전이하여 rhodomycin D를 생산하는 연구를 수행하였다. S. peucetius 유래의 7개 유전자 dnmU, T, J, V, Z, Q, S.를 당 합성 및 전이를 위해 plasmid 형태로 전이하였으며, S. venezuelae의 desIII, IV와 doxorubicin 내성 유전자인 drrA, B는 chromosomal DNA에 삽입하였다. Aglycone 기질인 ${\varepsilon}$-rhodomycinone을 확보하기 위하여 6L의 고체 배지에 S. peucetius를 배양하여 유기용매로 추출하고 preparative HPLC로 분리 정제하였다. 결과적으로 이종균주인 S. venezuelae에서 ${\varepsilon}$-rhodomycinone에 당 전이가 일어난 생산물을 확인함으로써 deoxy-sugar의 생합성 및 전이에 필요한 최소한의 유전적 정보를 확인할 수 있었다. 또한, 유사서열 단백질 모델링을 통하여, 최초로 당 전이 반응에 필수적인 도움효소 DnrQ의 구조를 예측하였다.

Enhanced Flavonoid Production in Streptomyces venezuelae via Metabolic Engineering

  • Park, Sung-Ryeol;Ahn, Mi-Sun;Han, Ah-Reum;Park, Je-Won;Yoon, Yeo-Joon
    • Journal of Microbiology and Biotechnology
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    • 제21권11호
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    • pp.1143-1146
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    • 2011
  • Metabolic engineering of plant-specific phenylpropanoid biosynthesis has attracted an increasing amount of attention recently, owing to the vast potential of flavonoids as nutraceuticals and pharmaceuticals. Recently, we have developed a recombinant Streptomyces venezuelae as a heterologous host for the production of flavonoids. In this study, we successfully improved flavonoid production by expressing two sets of genes predicted to be involved in malonate assimilation. The introduction of matB and matC encoding for malonyl-CoA synthetase and the putative dicarboxylate carrier protein, respectively, from Streptomyces coelicolor into the recombinant S. venezuelae strains expressing flavanone and flavone biosynthetic genes resulted in enhanced production of both flavonoids.

Identification and Functional Characterization of an afsR Homolog Regulatory Gene from Streptomyces venezuelae ATCC 15439

  • Maharjan, Sushila;Oh, Tae-Jin;Lee, Hei-Chan;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • 제19권2호
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    • pp.121-127
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    • 2009
  • Sequencing analysis of a 5-kb DNA fragment from Streptomyces venezuelae ATCC 15439 revealed the presence of one 3.1-kb open reading frame(ORF), designated as afsR-sv. The deduced product of afsR-sv(1,056 aa) was found to have high homology with the global regulatory protein AfsR. Homology-based analysis showed that aftR-sv represents a transcriptional activator belonging to the Streptomyces antibiotic regulatory protein(SARP) family that includes an N-terminal SARP domain containing a bacterial transcriptional activation domain(BTAD), an NB-ARC domain, and a C-terminal tetratricopeptide repeat domain. Gene expression analysis by reverse transcriptase PCR(RT-PCR) demonstrated the activation of transcription of genes belonging to pikromycin production, when aftR-sv was overexpressed in S. venezuelae. Heterologous expression of the aftR-sv in different Streptomyces strains resulted in increased production of the respective antibiotics, suggesting that afsR-sv is a positive regulator of antibiotics biosynthesis.

Interspecies Complementation of the LuxR Family Pathway-Specific Regulator Involved in Macrolide Biosynthesis

  • Mo, SangJoon;Yoon, Yeo Joon
    • Journal of Microbiology and Biotechnology
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    • 제26권1호
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    • pp.66-71
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    • 2016
  • PikD is a widely known pathway-specific regulator for controlling pikromycin production in Streptomyces venezuelae ATCC 15439, which is a representative of the large ATP-binding regulator of the LuxR family (LAL) in Streptomyces sp. RapH and FkbN also belong to the LAL family of transcriptional regulators, which show greatest homology with the ATP-binding motif and helix-turn-helix DNA-binding motif of PikD. Overexpression of pikD and heterologous expression of rapH and fkbN led to enhanced production of pikromycin by approximately 1.8-, 1.6-, and 1.6-fold in S. venezuelae, respectively. Cross-complementation of rapH and fkbN in the pikD deletion mutant (ΔpikD) restored pikromycin and derived macrolactone production. Overall, these results show that heterologous expression of rapH and fkbN leads to the overproduction of pikromycin and its congeners from the pikromycin biosynthetic pathway in S. venezuelae, and they have the same functionality as the pathwayspecific transcriptional activator for the pikromycin biosynthetic pathway in the ΔpikD strain. These results also show extensive "cross-communication" between pathway-specific regulators of streptomycetes and suggest revision of the current paradigm for pathwayspecific versus global regulation of secondary metabolism in Streptomyces species.

Heterologous Production and Detection of Recombinant Directing 2-Deoxystreptamine (DOS) in the Non-Aminoglycoside-Producing Host Streptomyces venezuelae YJ003

  • Kurumbang, Nagendra Prasad;Oh, Tae-Jin;Liou, Kwangkyoung;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • 제18권5호
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    • pp.866-873
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    • 2008
  • 2-Deoxystreptamine is a core aglycon that is vital to backbone formation in various aminoglycosides. This core structure can be modified to develop hybrid types of aminoglycoside antibiotics. We obtained three genes responsible for 2-deoxystreptamine production, neo7, neo6, and neo5, which encode 2-deoxy-scyllo-inosose synthase, L-glutamine: 2-deoxy-scyllo-inosose aminotransferase, and dehydrogenase, respectively, from the neomycin gene cluster. These genes were cloned into pIBR25, a Streptomyces expression vector, resulting in pNDOS. The recombinant pNDOS was transformed into a non-aminoglycoside-producing host, Streptomyces venezuelae YJ003, for heterologous expression. Based on comparisons of the retention time on LC-ESI/MS and ESI-MS data with those of the 2-deoxystreptamine standard, a compound produced by S. venezuelae YJ003/pNDOS was found to be 2-deoxystreptamine.

Biosynthesis of Glycosylated Derivatives of Tylosin in Streptomyces venezuelae

  • Han, Ah-Reum;Park, Sung-Ryeol;Park, Je-Won;Lee, Eun-Yeol;Kim, Dong-Myung;Kim, Byung-Gee;Yoon, Yeo-Joon
    • Journal of Microbiology and Biotechnology
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    • 제21권6호
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    • pp.613-616
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    • 2011
  • Streptomyces venezuelae YJ028, bearing a deletion of the entire biosynthetic gene cluster encoding the pikromycin polyketide synthases and desosamine biosynthetic enzymes, was used as a bioconversion system for combinatorial biosynthesis of glycosylated derivatives of tylosin. Two engineered deoxysugar biosynthetic pathways for the biosynthesis of TDP-3-O-demethyl-D-chalcose or TDP-L-rhamnose in conjunction with the glycosyltransferaseauxiliary protein pair DesVII/DesVIII were expressed in a S. venezuelae YJ028 mutant strain. Supplementation of each mutant strain capable of producing TDP-3-O-demethyl-D-chalcose or TDP-L-rhamnose with tylosin aglycone tylactone resulted in the production of the 3-O-demethyl-D-chalcose, D-quinovose, or L-rhamnose-glycosylated tylactone.

Genetically Engineered Biosynthesis of Macrolide Derivatives Including 4-Amino-4,6-Dideoxy-L-Glucose from Streptomyces venezuelae YJ003-OTBP3

  • Pageni, Binod Babu;Oh, Tae-Jin;Liou, Kwang-Kyoung;Yoon, Yeo-Joon;Sohng, Jae-Kyung
    • Journal of Microbiology and Biotechnology
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    • 제18권1호
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    • pp.88-94
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    • 2008
  • Two sugar biosynthetic cassette plasm ids were used to direct the biosynthesis of a deoxyaminosugar. The pOTBP1 plasmid containing TDP-glucose synthase (desIII), TDP-glucose-4,6-dehydratase (desIV), and glycosyltransferase (desVII/desVIII) was constructed and transformed into S. venezuelae YJ003, a strain in which the entire gene cluster of desosamine biosynthesis is deleted. The expression plasmid pOTBP3 containing 4-aminotransferase (gerB) and 3,5-epimerase (orf9) was transformed again into S. venezuelae YJ003-OTBP1 to obtain S. venezuelae YJ003-OTBP3 for the production of 4-amino-4,6-dideoxy-L-glucose derivatives. The crude extracts obtained from S. venezuelae ATCC 15439, S. venezuelae YJ003, and S. venezuelae YJ003-OTBP3 were further analyzed by TLC, bioassay, HPLC, ESI/MS, LC/MS, and MS/MS. The results of our study clearly shows that S. venezuelae YJ003-OTBP3 constructs other new hybrid macrolide derivatives including 4-amino-4,6-dideoxy-L-glycosylated YC-17 (3, [M+ $Na^+$] m/z=464.5), methymycin (4, m/z=480.5), novamethymycin (6, m/z=496.5), and pikromycin (5, m/z=536.5) from a 12-membered ring aglycon (10-deoxymethynolide, 1) and a 14-membered ring aglycon (narbonolide, 2). These results suggest a successful engineering of a deoxysugar pathway to generate novel hybrid macrolide derivatives, including deoxyaminosugar.

Hydroxylation of Indole by PikC Cytochrome P450 from Streptomyces venezuelae and Engineering Its Catalytic Activity by Site-Directed Mutagenesis

  • Lee Sang-Kil;Park Je-Won;Park Sung-Ryeol;Ahn Jong-Seog;Choi Cha-Yong;Yoon Yeo-Joon
    • Journal of Microbiology and Biotechnology
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    • 제16권6호
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    • pp.974-978
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    • 2006
  • The cytochrome P450 monooxygenase from the pikromycin biosynthetic gene cluster in Streptomyces venezuelae, known as PikC, was observed to hydroxylate the unnatural substrate indole to indigo. Furthermore, the site-directed mutagenesis of PikC monooxygenase led to the mutant enzyme F171Q, in which Phe171 was altered to Gln, with enhanced activity for the hydroxylation of indole. From enzyme kinetic studies, F171Q showed an approximately five-fold higher catalytic efficiency compared with the wild-type PikC. Therefore, these results demonstrate the promising application of P450s originating from Streptomyces, normally involved in polyketide biosynthesis, to generate a diverse array of other industrially useful compounds.