title: Genome-engineered Sinorhizobium meliloti for the production of poly(lactic-co-3-hydroxybutyric) acid copolymer.
abstract: Economically competitive commercial production of biodegradable bioplastics with desirable properties is an important goal. In this study, we demonstrate the use of chromosome engineering of an alternative bacterial host, Sinorhizobium meliloti, for production of the copolymer, poly(lactate-co-3-hydroxybutyrate). Codon-optimized genes for 2 previously engineered enzymes, Clostridium propionicum propionate CoA transferase (Pct532Cp) and Pseudomonas sp. strain MBEL 6-19 polyhydroxyalkanoate (PHA) synthase 1 (PhaC1400Ps6-19), were introduced into S. meliloti Rm1021 by chromosome integration, replacing the native phbC gene. On the basis of phenotypic analysis and detection of polymer product by gas chromatography analysis, synthesis and accumulation of the copolymer was confirmed. The chromosome integrant strain, with the introduced genes under the control of the native phbC promoter, is able to produce over 15% cell dry mass of poly(lactate-co-3-hydroxybutyrate), containing 30 mol% lactate, from growth on mannitol. We were also able to purify the polymer from the culture and confirm the structure by NMR and GC-MS. To our knowledge, this is the first demonstration of production of this copolymer in the Alphaproteobacteria. Further optimization of this system may eventually yield strains that are able to produce economically viable commercial product.
authors.affiliation: Department of Biology, Faculty of Science, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada.|Department of Biology, Faculty of Science, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada.
authors.forename: Tam T
authors.initials: TT
authors.lastname: Tran
authors.affiliation: Department of Biology, Faculty of Science, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada.|Department of Biology, Faculty of Science, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada.
authors.forename: Trevor C
authors.initials: TC
authors.lastname: Charles
mesh_terms: D000217:Acyltransferases / Q000235:genetics*; D015256:Coenzyme A-Transferases / Q000235:genetics*; D005818:Genetic Engineering; D006885:Hydroxybutyrates / Q000378:metabolism; D011091:Polyesters / Q000378:metabolism*; D016962:Sinorhizobium meliloti / Q000235:genetics*; D000094122:Polyhydroxybutyrates
chemical_list: D000217:Acyltransferases; D015256:Coenzyme A-Transferases; D006885:Hydroxybutyrates; D011091:Polyesters; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C033616:poly(lactide); C077225:poly(3-hydroxyalkanoic acid) synthase; C031212:acyl-CoA transferase
keywords: PHA synthase; Sinorhizobium meliloti génétiquement conçu; bioplastics; bioplastiques; copolymer; copolymère; engineered Sinorhizobium meliloti; propionate CoA transferase; propionate CoA transférase