Open AccessHarvesting of novel polyhydroxyalkanaote (PHA) synthase encoding genes from a soil metagenome library using phenotypic screening
Author(s) -
Schallmey Marcus,
Ly Anh,
Wang Chunxia,
Meglei Gabriela,
Voget Sonja,
Streit Wolfgang R.,
Driscoll Brian T.,
Charles Trevor C.
Publication year - 2011
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2011.02324.x
Subject(s) - complementation , cosmid , biology , gene , metagenomics , sinorhizobium meliloti , genetics , mutant , genomic library , computational biology , phenotype , phenotypic screening , peptide sequence
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.