Title | In vitro analysis of the chain termination reaction in the synthesis of poly-(R)-beta-hydroxybutyrate by the class III synthase from Allochromatium vinosum. |
Publication Type | Journal Article |
Year of Publication | 2005 |
Authors | Lawrence, AG, Choi, J, Rha, CK, Stubbe, J, Sinskey, AJ |
Journal | Biomacromolecules |
Volume | 6 |
Issue | 4 |
Pagination | 2113-9 |
Date Published | 2005 Jul-Aug |
ISSN | 1525-7797 |
Keywords | Acyltransferases, Chromatiaceae, Escherichia coli, Hydroxybutyrates, Magnetic Resonance Spectroscopy, Molecular Weight, Polyesters |
Abstract | Allochromatium vinosum polyhydroxyalkanoate synthase catalyzes formation of poly-(R)-3-hydroxybutyrate (PHB) from (R)-3-hydroxybutyryl-coenzyme A (HB-CoA). (R)-3-Hydroxybutyryl-N-acetylcysteamine (HB-NAC) is an alternative substrate for this synthase in vitro, with a turnover 1% that of HB-CoA. With HB-NAC, the molecular weight (M(w)) of PHB produced at substrate-to-enzyme ratios of 1500-15 000 is approximately 75 kDa. (1)H NMR shows that PHB produced has hydroxybutyrate at the alcohol end and N-acetylcysteamine (NAC) at the carboxylate end of the polymer. Exogenous NAC has no effect on the M(w) of PHB produced with HB-CoA or HB-NAC in vitro, whereas PHB from a polymerization reaction with both HB-CoA and HB-NAC has intermediate M(w). These results can be accommodated by two models. In the first, NAC liberated at the active site during polymerization acts as a chain transfer agent. In the second, there is a noncovalent polymer intermediate covalently linked to NAC, which can dissociate from the active site. |
DOI | 10.1021/bm0501048 |
Alternate Journal | Biomacromolecules |
Citation Key | 80 |
PubMed ID | 16004452 |
Grant List | 49171 / / PHS HHS / United States |