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Acylation of Streptomyces type II polyketide synthase acyl carrier proteins
Author(s) -
Crosby John,
Byrom Kate J,
Hitchman Timothy S,
Cox Russell J,
Crump Matthew P,
Findlow I.Stuart C,
Bibb Maureen J,
Simpson Thomas J
Publication year - 1998
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(98)00840-0
Subject(s) - acylation , acyl carrier protein , polyketide , chemistry , cysteine , stereochemistry , streptomyces , intramolecular force , biochemistry , residue (chemistry) , polyketide synthase , biosynthesis , enzyme , biology , bacteria , genetics , catalysis
Acyl derivatives of type II PKS ACPs are required for in vitro studies of polyketide biosynthesis. The presence of an exposed cysteine residue prevented specific chemical acylation of the phosphopantetheine thiol of the actinorhodin PKS holo ACP. Acylation studies were further complicated by intramolecular disulphide formation between cysteine 17 and the phosphopantetheine. The presence of this intramolecular disulphide was confirmed by tryptic digestion of the ACP followed by ESMS analysis of the fragments. An act Cys17Ser ACP was engineered by site‐directed mutagenesis. S ‐Acyl adducts of act C17S, oxytetracycline and griseusin holo ACPs were rapidly formed by reaction with hexanoyl, 5‐ketohexanoyl and protected acetoacetyl imidazolides. Comparisons with type II FAS ACPs were made.