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Volume 147, Issue 6

Cloning and functional analysis of a phosphopantetheinyl transferase superfamily gene associated with jadomycin biosynthesis in ISP5230

The GenBank accession numbers for the and sequences reported in this paper are AF222693 and AY026363, respectively.

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Abstract

Sequence analysis of a I/I fragment of chromosomal DNA downstream of in the ISP5230 gene cluster for jadomycin biosynthesis detected a partial ORF similar in its deduced amino acid sequence to the product involved in synthesizing a regulator of heterocyst spacing in . By probing a phage library of DNA with the I/I fragment, the authors identified and isolated a hybridizing clone. The nucleotide sequence of its DNA contained three complete ORFs (, and ) and one incomplete ORF (). The ORF lay immediately downstream of, and partially overlapped, . It contained 786 nucleotides encoding an amino acid sequence like those of enzymes in the phosphopantetheinyl transferase family. The ORF contained 1794 nucleotides and encoded an amino acid sequence resembling acyl-CoA decarboxylases, thus suggesting a role in polyketide condensation reactions.The ORF was not identified, but the partial showed marked similarities in its deduced amino acid sequence to NDP-hexose-2,3-dehydratases, indicating a role in forming the sugar component of jadomycin B. Expression of in and examination of the product by SDS-PAGE established that the ORF encoded a 291 kDa protein, corresponding in size to the 262 amino acid polypeptide deduced from the sequence. Evidence from a Northern hybridization indicated that expression is correlated with jadomycin B synthesis. Cultures of ISP5230 disrupted in produced only 2–5% of the wild-type titre of jadomycin B, but grew well and produced chloramphenicol normally. The authors conclude that encodes a holo-ACP synthase needed primarily for jadomycin B biosynthesis.

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Keyword(s): ACP, acyl carrier protein , ACPS, holo-acyl carrier protein synthase , act, actinorhodin biosynthesis gene , acyl carrier protein synthase , ArCP, aryl carrier protein , CLF, chain-length factor , cml, chloramphenicol biosynthesis gene , fren, frenolicin biosynthesis gene , gra, granaticin biosynthesis gene , jad genes , jad, jadomycin biosynthesis gene , KS, ketosynthase , NRPS, non-ribosomal peptide synthetase , PCP, peptidyl carrier protein , phosphopantetheinyl transferase gene , PKS, polyketide synthase , polyketide antibiotic , Ppan, phosphopantetheine and PPTase, phosphopantetheinyl transferase
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2001-06-01
2024-05-03
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Cloning and functional analysis of a phosphopantetheinyl transferase superfamily gene associated with jadomycin biosynthesis in Streptomyces venezuelae ISP5230The GenBank accession numbers for the jadM and jadNOX sequences reported in this paper are AF222693 and AY026363, respectively.
Microbiology 147, 1535 (2001); https://doi.org/10.1099/00221287-147-6-1535
/content/journal/micro/10.1099/00221287-147-6-1535
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