title: Harvesting of novel polyhydroxyalkanaote (PHA) synthase encoding genes from a soil metagenome library using phenotypic screening.
abstract: We previously reported the construction of metagenomic libraries in the IncP cosmid vector pRK7813, enabling heterologous expression of these broad-host-range libraries in multiple bacterial hosts. Expressing these libraries in Sinorhizobium meliloti, we have successfully complemented associated phenotypes of polyhydroxyalkanoate synthesis mutants. DNA sequence analysis of three clones indicates that the complementing genes are homologous to, but substantially different from, known polyhydroxyalkanaote synthase-encoding genes. Thus we have demonstrated the ability to isolate diverse genes for polyhydroxyalkanaote synthesis by functional complementation of defined mutants. Such genes might be of use in the engineering of more efficient systems for the industrial production of bioplastics. The use of functional complementation will also provide a vehicle to probe the genetics of polyhydroxyalkanaote metabolism and its relation to carbon availability in complex microbial assemblages.
authors.affiliation: Department of Biology, University of Waterloo, Waterloo, ON, Canada.
authors.forename: Marcus
authors.initials: M
authors.lastname: Schallmey
authors.forename: Anh
authors.initials: A
authors.lastname: Ly
authors.forename: Chunxia
authors.initials: C
authors.lastname: Wang
authors.forename: Gabriela
authors.initials: G
authors.lastname: Meglei
authors.forename: Sonja
authors.initials: S
authors.lastname: Voget
authors.forename: Wolfgang R
authors.initials: WR
authors.lastname: Streit
authors.forename: Brian T
authors.initials: BT
authors.lastname: Driscoll
authors.forename: Trevor C
authors.initials: TC
authors.lastname: Charles
mesh_terms: D000217:Acyltransferases / Q000235:genetics*; D003001:Cloning, Molecular / Q000379:methods*; D015723:Gene Library; D005816:Genetic Complementation Test; D054892:Metagenome / Q000235:genetics*; D046529:Microscopy, Electron, Transmission; D005810:Multigene Family; D009154:Mutation; D010641:Phenotype; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D016962:Sinorhizobium meliloti; D012988:Soil Microbiology
chemical_list: D000217:Acyltransferases; C077225:poly(3-hydroxyalkanoic acid) synthase