title: Biosynthesis of glycolate-based polyesters containing medium-chain-length 3-hydroxyalkanoates in recombinant Escherichia coli expressing engineered polyhydroxyalkanoate synthase.
abstract: Glycolate(GL)-based polyesters were for the first time produced in the recombinant Escherichia coli fatty acid β-oxidation pathway reinforcing mutant LS5218, using extracellularly added GL as a monomer precursor. Cells expressing a Ser325Thr/Gln481Lys mutant of polyhydroxyalkanoate synthase (PhaC1STQK) from Pseudomonas sp. 61-3, propionyl-CoA transferase from Megasphaera elsdenii and enoyl-CoA hydratase from Pseudomonas aeruginosa grown on GL and dodecanoate were found to produce novel copolymers of GL with 3-hydroxyalkanoates (3HAs) (C(4)-C(12)), P(GL-co-3HA), with a weight-average molecular weight of 34,000. The (1)H and (13)C NMR analyses of the copolymer revealed the incorporation of GL units into the polymer chain. This result demonstrates that PhaC1STQK polymerized glycolyl-CoA as a monomer substrate. Additionally, the novel lactate(LA)-based polyester P(LA-co-3HA) was produced by substituting GL with LA, indicating that the method is versatile and allows the production of a variety of biopolymers.
authors: Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13-W8, Kita-ku, Sapporo 060-8628, Japan. mken@eng.hokudai.ac.jp
authors: Ken'ichiro
authors: K
authors: Matsumoto
authors: Ayako
authors: A
authors: Ishiyama
authors: Kohei
authors: K
authors: Sakai
authors: Tetsufumi
authors: T
authors: Shiba
authors: Seiichi
authors: S
authors: Taguchi
mesh_terms: D000217:Acyltransferases / Q000096:biosynthesis / Q000235:genetics; D057005:Bioengineering; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial; D006016:Glycolates / Q000378:metabolism; D011091:Polyesters / Q000378:metabolism; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism
chemical_list: D006016:Glycolates; D011091:Polyesters; C031149:glycolic acid; D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase