Ablation of the OtcC gene encoding a post-polketide hydroxylase from the oxytetracyline biosynthetic pathway in streptomyces rimosus results in novel polyketides with altered chain length

Nataša Peric-Concha, Branko Borovicka, Paul F. Long, Daslav Hranueli, Peter G. Waterman, Iain S. Hunter

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Oxytetracycline (OTC) is a 19-carbon polyketide antibiotic made by Streptomyces rimosus. The otcC gene encodes an anhydrotetracycline oxygenase that catalyzes a hydroxylation of the anthracycline structure at position C-6 after biosynthesis of the polyketide backbone is completed. A recombinant strain of S. rimosus that was disrupted in the genomic copy of otcC synthesized a novel C-17 polyketide. This result indicates that the absence of the otcC gene product significantly influences the ability of the OTC “minimal” polyketide synthase to make a polyketide product of the correct chain length. A mutant copy of otcC was made by site-directed mutagenesis of three essential glycine codons located within the putative NADPH-binding domain. The mutant gene was expressed in Escherichia coli, and biochemical analysis confirmed that the gene product was catalytically inactive. When the mutant gene replaced the ablated gene in the chromosome of S. rimosus, the ability to make a 19-carbon backbone was restored, indicating that OtcC is an essential partner in the quaternary structure of the synthase complex.
LanguageEnglish
Pages37455-37460
Number of pages6
JournalJournal of Biological Chemistry
Volume280
DOIs
Publication statusPublished - 11 Nov 2005

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Streptomyces rimosus
Polyketides
Gene encoding
Biosynthetic Pathways
Mixed Function Oxygenases
Ablation
Chain length
Genes
Oxytetracycline
Carbon
Polyketide Synthases
Hydroxylation
Mutagenesis
Anthracyclines
Biosynthesis
Chromosomes
Site-Directed Mutagenesis
NADP
Codon
Glycine

Keywords

  • oxytetracycline
  • ablated gene
  • synthase complex

Cite this

Peric-Concha, Nataša ; Borovicka, Branko ; Long, Paul F. ; Hranueli, Daslav ; Waterman, Peter G. ; Hunter, Iain S. / Ablation of the OtcC gene encoding a post-polketide hydroxylase from the oxytetracyline biosynthetic pathway in streptomyces rimosus results in novel polyketides with altered chain length. In: Journal of Biological Chemistry. 2005 ; Vol. 280. pp. 37455-37460.
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abstract = "Oxytetracycline (OTC) is a 19-carbon polyketide antibiotic made by Streptomyces rimosus. The otcC gene encodes an anhydrotetracycline oxygenase that catalyzes a hydroxylation of the anthracycline structure at position C-6 after biosynthesis of the polyketide backbone is completed. A recombinant strain of S. rimosus that was disrupted in the genomic copy of otcC synthesized a novel C-17 polyketide. This result indicates that the absence of the otcC gene product significantly influences the ability of the OTC “minimal” polyketide synthase to make a polyketide product of the correct chain length. A mutant copy of otcC was made by site-directed mutagenesis of three essential glycine codons located within the putative NADPH-binding domain. The mutant gene was expressed in Escherichia coli, and biochemical analysis confirmed that the gene product was catalytically inactive. When the mutant gene replaced the ablated gene in the chromosome of S. rimosus, the ability to make a 19-carbon backbone was restored, indicating that OtcC is an essential partner in the quaternary structure of the synthase complex.",
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Ablation of the OtcC gene encoding a post-polketide hydroxylase from the oxytetracyline biosynthetic pathway in streptomyces rimosus results in novel polyketides with altered chain length. / Peric-Concha, Nataša; Borovicka, Branko; Long, Paul F.; Hranueli, Daslav; Waterman, Peter G.; Hunter, Iain S.

In: Journal of Biological Chemistry, Vol. 280, 11.11.2005, p. 37455-37460.

Research output: Contribution to journalArticle

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T1 - Ablation of the OtcC gene encoding a post-polketide hydroxylase from the oxytetracyline biosynthetic pathway in streptomyces rimosus results in novel polyketides with altered chain length

AU - Peric-Concha, Nataša

AU - Borovicka, Branko

AU - Long, Paul F.

AU - Hranueli, Daslav

AU - Waterman, Peter G.

AU - Hunter, Iain S.

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N2 - Oxytetracycline (OTC) is a 19-carbon polyketide antibiotic made by Streptomyces rimosus. The otcC gene encodes an anhydrotetracycline oxygenase that catalyzes a hydroxylation of the anthracycline structure at position C-6 after biosynthesis of the polyketide backbone is completed. A recombinant strain of S. rimosus that was disrupted in the genomic copy of otcC synthesized a novel C-17 polyketide. This result indicates that the absence of the otcC gene product significantly influences the ability of the OTC “minimal” polyketide synthase to make a polyketide product of the correct chain length. A mutant copy of otcC was made by site-directed mutagenesis of three essential glycine codons located within the putative NADPH-binding domain. The mutant gene was expressed in Escherichia coli, and biochemical analysis confirmed that the gene product was catalytically inactive. When the mutant gene replaced the ablated gene in the chromosome of S. rimosus, the ability to make a 19-carbon backbone was restored, indicating that OtcC is an essential partner in the quaternary structure of the synthase complex.

AB - Oxytetracycline (OTC) is a 19-carbon polyketide antibiotic made by Streptomyces rimosus. The otcC gene encodes an anhydrotetracycline oxygenase that catalyzes a hydroxylation of the anthracycline structure at position C-6 after biosynthesis of the polyketide backbone is completed. A recombinant strain of S. rimosus that was disrupted in the genomic copy of otcC synthesized a novel C-17 polyketide. This result indicates that the absence of the otcC gene product significantly influences the ability of the OTC “minimal” polyketide synthase to make a polyketide product of the correct chain length. A mutant copy of otcC was made by site-directed mutagenesis of three essential glycine codons located within the putative NADPH-binding domain. The mutant gene was expressed in Escherichia coli, and biochemical analysis confirmed that the gene product was catalytically inactive. When the mutant gene replaced the ablated gene in the chromosome of S. rimosus, the ability to make a 19-carbon backbone was restored, indicating that OtcC is an essential partner in the quaternary structure of the synthase complex.

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