|Ashby, Richard - Rick|
Submitted to: Biomacromolecules
Publication Type: Review Article
Publication Acceptance Date: 11/18/2004
Publication Date: 5/1/2005
Citation: Solaiman, D., Ashby, R.D. 2005. Rapid genetic characterization of poly(hydroxyalkanoate) synthase and its applications. Biomacromolecules. 6(2):532-537.
Technical Abstract: Microorganisms containing short-chain-length (scl-) or medium-chain-length (mcl-) poly(hydroxyalkanoates) (PHA) are commonly identified by staining methods using lipophilic reagents. These methods provide powerful means for general screening of organisms actively producing and accumulating PHAs. The Southern blot hybridization method additionally allows the identification of potential PHA-producing microorganisms. PCR-based detection methods further afford rapid and sensitive means to screen for PHA biosynthesis genes. We have developed PCR assays for simultaneous or individual detection of the mcl-PHA synthase genes of Pseudomonas. The amplicons (~ 0.5 kb) can be directly sequenced or used as probes for hybridization studies. The sequence information can further be used to initiate chromosome walking for eventual cloning of the complete PHA biosynthesis operon. We have also used amplification pattern and sequence data to differentiate sub-groups of P. corrugata and P. mediterranea. Other researchers reported PCR methods for the detection of scl-PHA synthase genes and those of Bacillus spp., thus greatly expanding the types of PHA synthase gene and the organisms that can be characterized by this approach. The vast sequence information obtainable through PCR-based studies of various PHA synthase operons should facilitate the identification or construction of new PHA synthases capable of synthesizing novel PHAs.