• Title/Summary/Keyword: polyketide synthase

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Characterization of Doxorubicin-nonproducing Mutant, Nu3 of Streptomyces peucetius ATCC27952

  • Kyu, Hwang-Cheol;Lee, Hong-Sub;Hong, Young-Soo;Paek, Nam-Soo;Kim, Tae-Han;Lee, Jung-Joon
    • Journal of Microbiology and Biotechnology
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    • v.7 no.5
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    • pp.363-366
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    • 1997
  • A doxorubicin-nonproducing mutant, Nu23 was selected from the mutagenesis of Streptomyces peucetius ATCC27952. Chemical and molecular biological analysis suggested that the mutant was blocked at the step between polyketide synthase and aklaviketon reductase in the biosynthesis of doxorubicin. Furthermore, the bioconversion experiment with the mutant revealed that 13-dihydrodaunorubicin is likely to be a biosynthetic intermediate.

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Comparative Transcriptome Analysis for Avermectin Overproduction via Streptomyces avermitilis Microarray System

  • Im, Jong-Hyuk;Kim, Myung-Gun;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.3
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    • pp.534-538
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    • 2007
  • Avermectin and its analogs are major commercial antiparasitic agents in the fields of animal health, agriculture, and human infections. To increase our understanding about the genetic mechanism underlying avermectin overproduction, comparative transcriptomes were analyzed between the low producer S. avermitilis ATCC31267 and the high producer S. avermitilis ATCC31780 via a S. avermitilis whole genome chip. The comparative transcriptome analysis revealed that fifty S. avermitilis genes were expressed at least two-fold higher in S. avermitilis ATCC31780. In particular, all the avermectin biosynthetic genes, including polyketide synthase (PKS) genes and an avermectin pathway-specific regulatory gene, were less expressed in the low producer S. avermitilis ATCC31267. The present results imply that avermectin overproduction in S. avermitilis ATCC31780 could be attributed to the previously unidentified fifty genes reported here and increased transcription levels of avermectin PKS genes.

Identification of Three Positive Regulators in the Geldanamycin PKS Gene Cluster of Streptomyces hygroscopicus JCM4427

  • Kim, Won-Cheol;Lee, Jung-Joon;Paik, Sang-Gi;Hong, Young-Soo
    • Journal of Microbiology and Biotechnology
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    • v.20 no.11
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    • pp.1484-1490
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    • 2010
  • In the Streptomyces hygroscopicus JCM4427 geldanamycin biosynthetic gene cluster, five putative regulatory genes were identified by protein homology searching. Among those genes, gel14, gel17, and gel19 are located downstream of polyketide synthase genes. Gel14 and Gel17 are members of the LAL family of transcriptional regulators, including an ATP/GTP-binding domain at the N-terminus and a DNA-binding helix-turn-helix domain at the C-terminus. Gel19 is a member of the TetR family of transcriptional regulators, which generally act to repress transcription. To verify the biological significance of the putative regulators in geldanamycin production, they were individually characterized by gene disruption, genetic complementation, and transcriptional analyses. All three genes were confirmed as positive regulators of geldanamycin production. Specifically, Gel17 and Gel19 are required for gel14 as well as gelA gene expression.

Characterization of a Chalcosyltransferase (gerGTII) in Dihydrochalcomycin Biosynthesis

  • Pageni, Binod Babu;Oh, Tae-Jin;Thuy, Ta Thi Thu;Sohng, Jae Kyung
    • Molecules and Cells
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    • v.26 no.3
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    • pp.278-284
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    • 2008
  • An open reading frame, designated GerGTII and located downstream of the polyketide synthase genes, has been identified as a chalcosyltransferase by sequence analysis in the dihydrochalcomycin biosynthetic gene cluster of Streptomyces sp. KCTC 0041BP. The deduced product of gerGTII is similar to several glycosyltransferases, authentic and putative, and it displays a consensus sequence motif that appears to be characteristic of a sub-group of these enzymes. Specific disruption of gerGTII within the S. sp. KCTC 0041BP genome by insertional in-frame deletion method, resulted complete abolishment of dihydrochalcomycin and got the 20-O-mycinosyl-dihydrochalconolide as intermediate product in dihydrochalcomycin biosynthesis which was confirmed by electron spray ionization-mass spectrometry and liquid chromatography-mass spectrometry. Dihydrochalcomycin also was recovered after complementation of gerGTII.

Draft Genome Sequence of Weissella koreensis Strain HJ, a Probiotic Bacterium Isolated from Kimchi

  • Seung-Min Yang;Eiseul Kim;So-Yun Lee;Soyeong Mun;Hae Choon Chang;Hae-Yeong Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.128-131
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    • 2023
  • Here we report the draft genome sequence of Weissella koreensis strain HJ and genomic analysis of its key features. The genome consists of 1,427,571 bp with a GC content of 35.5%, and comprises 1,376 coding genes. In silico analysis revealed the absence of pathogenic factors within the genome. The genome harbors several genes that play an important role in the survival of the gastrointestinal tract. In addition, a type III polyketide synthase cluster was identified. Pangenome analysis identified 68 unique genes in W. koreensis strain HJ. The genome information of this strain provides the basis for understanding its probiotic properties.

Eicosapentaenoic Acid (EPA) Biosynthetic Gene Cluster of Shewanella oneidensis MR-1: Cloning, Heterologous Expression, and Effects of Temperature and Glucose on the Production of EPA in Escherichia coli

  • Lee, Su-Jin;Jeong, Young-Su;Kim, Dong-Uk;Seo, Jeong-Woo;Hur, Byung-Ki
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.6
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    • pp.510-515
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    • 2006
  • The putative EPA synthesis gene cluster was mined from the entire genome sequence of Shewanella oneidensis MR-1. The gene cluster encodes a PKS-like pathway that consists of six open reading frames (ORFs): ORFSO1602 (multi-domain beta-ketoacyl synthase, KS-MAT-4ACPs-KR), ORFSO1600 (acyl transferase, AT), ORFSO1599 (multi-domain beta-ketoacyl synthase, KS-CLF-DH-DH), ORFSO1597 (enoyl reductase, ER), ORFSO1604 (phosphopentetheine transferase, PPT), and ORFSO1603 (transcriptional regulator). In order to prove involvement of the PKS-like machinery in EPA synthesis, a 20.195-kb DNA fragment containing the genes was amplified from S. oneidensis MR-1 by the long-PCR method. Its identity was confirmed by the methods of restriction enzyme site mapping and nested PCR of internal genes orfSO1597 and orfSO1604. The DNA fragment was cloned into Escherichia coli using cosmid vector SuperCos1 to form pCosEPA. Synthesis of EPA was observed in four E. coli clones harboring pCosEPA, of which the maximum yield was 0.689% of the total fatty acids in a clone designated 9704-23. The production yield of EPA in the E. coli clone was affected by cultivation temperature, showing maximum yield at $20^{\circ}C$ and no production at $30^{\circ}C$ or higher. In addition, production yield was inversely proportional to glucose concentration of the cultivation medium. From the above results, it was concluded that the PKS-like modules catalyze the synthesis of EPA. The synthetic process appears to be subject to regulatory mechanisms triggered by various environmental factors. This most likely occurs via the control of gene expression, protein stability, or enzyme activity.

Molecular Classification of Commercial Spirulina Strains and Identification of Their Sulfolipid Biosynthesis Genes

  • Kwei, Chee Kuan;Lewis, David;King, Keith;Donohue, William;Neilan, Brett A.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.359-365
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    • 2011
  • Cyanobacterial strains of the genus Spirulina have recently been identified as an excellent source of sulfolipids, some of which possess anti-HIV properties. Thus, to investigate the distribution of sufolipid biosynthesis pathways in Spirulina, a genetic screening/phylogentic study was performed. Five different strains of Spirulina [Spirulina (Jiangmen), Spirulina sp., S. platensis, S. maxima, and Spirulina seawater] sourced from different locations were initially classified via 16S rDNA sequencing, and then screened for the presence of the sulfolipid biosynthesis genes sqdB and sqdX via a PCR. To assess the suitability of these strains for human consumption and safe therapeutic use, the strains were also screened for the presence of genes encoding nonribosomal peptide synthases (NRPSs) and polyketide synthases (PKSs), which are often associated with toxin pathways in cyanobacteria. The results of the 16S rDNA analysis and phylogenetic study indicated that Spirulina sp. is closely related to Halospirulina, whereas the other four Spirulina strains are closely related to Arthrospira. Homologs of sqdB and sqdX were identified in Spirulina (Jiangmen), Spirulina sp., S. platensis, and the Spirulina seawater. None of the Spirulina strains screened in this study tested positive for NRPS or PKS genes, suggesting that these strains do not produce NRP or PK toxins.

Fumonisin Production by Field Isolates of the Gibberella fujikuroi Species Complex and Fusarium commune Obtained from Rice and Corn in Korea (우리나라 벼와 옥수수로부터 분리한 Gibberella fujikuroi 종복합체와 Fusarium commune 소속 균주의 푸모니신 생성능)

  • Lee, Soo-Hyung;Kim, Ji-Hye;Son, Seung-Wan;Lee, Theresa;Yun, Sung-Hwan
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.310-316
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    • 2012
  • Gibberellea fujikuroi species (Gf) complex comprises at least 15 species, most of which not only causes serious plant diseases, but also produces mycotoxins including fumonisins. Here, we focused on the abilities of the field isolates belonging to the Gf complex associated with rice and corn, respectively in Korea to produce fumonisin, all of which were confirmed to carry FUM1, the polyketide synthase gene essential for fumonisin biosynthesis. A total of 88 Gf complex isolates (55 F. fujikuroi, 10 F. verticillioides, 20 F. proliferatum, 2 F. subglutinans, and 1 F. concentricum), and 4 isolates of F. commune, which is a non-member of Gf complex, were grown on rice substrate and determined for their production levels of fumonisins by a HPLC method. Most isolates of F. verticillioides and F. proliferatum, regardless of host origins, produced fumonisin $B_1$ and $B_2$ at diverse ranges of levels ($0.5-2,686.4{\mu}g/g$, and $0.7-1,497.6{\mu}g/g$, respectively). In contrast, all the isolates of F. fujikuroi and other Fusarium species examined produced no fumonisins or only trace amounts ($<10{\mu}g/g$) of fumonisins. Interestingly, the frequencies of relatively high fumonisin-producers among the F. proliferatum and F. fujikuroi isolates derived from corn were higher than those among the fungal isolates from rice. In addition, it is a first report demonstrating the ability of the FUM1-carrying F. commune isolates from rice to produce fumonisins.

Isolation and Characterization of Actinomycete Strain BK185 Possessing Antifungal Activity against Ginseng Root Rot Pathogens (인삼 뿌리썩음병균에 항균활성이 있는 방선균 BK185의 분리 및 특성)

  • Kim, Byung-Yong;Bae, Mun-Hyung;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Kyu;Oh, Dong-Chan;Song, Jaekyeong
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.396-403
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    • 2014
  • Ginseng (Panax ginseng C. A. Meyer) is an economically valuable pharmaceutical crop in Korea. In order to find promising biocontrol agents for soil-borne fungal pathogens which infect ginseng roots, we have isolated actinomycete, BK185 from soil. The isolate was investigated for the antifungal activity against to ginseng rot pathogens prior to testing genetic and chemical properties. The strain was identified as Streptomyces sp. using phylogenetic analysis based on 16S rRNA gene sequence. The most closely related species was S. sporoclivatus and S. geldanamycininus with high similarities (>99%). The isolate, BK185 showed positive reaction for PCR detection targeting biosynthetic gene clusters of PKS (Type-I polyketide synthase) and NRPS (Non-ribosomal polypeptide synthetase) genes. Major metabolite from the BK185 was analyzed by The LC/MS and identified to geldamycin, which was known to contained broad antibacterial, antifungal or anticancer activities. The results provide evidences that the strain, BK185 can be promising biocontrol agent for ginseng organic farming.

Isoflavone Daidzein: Chemistry and Bacterial Metabolism

  • Kim, Mi-Hyang;Han, Jae-Hong;Kim, Soo-Un
    • Journal of Applied Biological Chemistry
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    • v.51 no.6
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    • pp.253-261
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    • 2008
  • Isoflavone daidzein is a phytoestrogen widely distributed in Leguminosae and is especially rich in the soybean. The C6-C3 (rings B and C) unit of isoflavones is derived from the phenylpropanoid pathway and the remaining C6 (ring A) unit is from the polyketide pathway. This unique carbon skeleton is the result of isomerization of the flavone catalyzed by the isoflavone synthase, a cytochrome P450 enzyme. The isoflavones daidzein and genistein are present in the plant mostly in the glucosylated forms. However, in the human intestine, the glycosidic linkage is broken, and the free form is uptaked into blood stream. The free form is further metabolized into various reduction products to end up at the equol, which is known to have the most potent estrogenic effect among the metabolites. Several human intestinal bacteria that can convert daidzein into equol have been described, and the study into the chemistry and biochemistry of the daizein reduction would be rewarding to the improvement of the human health.