1887

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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2001-06-01
2021-07-30
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