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Title: Construction and Characterization of Fusion Class III Poly(hydroxyalkanoate) Biopolymer Synthase Genes

Author
item Solaiman, Daniel
item Ashby, Richard - Rick
item Zerkowski, Jonathan

Submitted to: Annual Meeting and Expo of the American Oil Chemists' Society
Publication Type: Abstract Only
Publication Acceptance Date: 1/13/2010
Publication Date: 5/2/2010
Citation: Solaiman, D., Ashby, R.D., Zerkowski, J.A. 2010. Construction and Characterization of Fusion Class III Poly(hydroxyalkanoate) Biopolymer Synthase Genes. Annual Meeting and Expo of the American Oil Chemists' Society Abstracts. p. 19.

Interpretive Summary:

Technical Abstract: Poly(hydroxyalkanoates) (PHAs) are polyesters produced by bacteria. Because these polymers are biodegradable and are produced using renewable fermentative substrates, PHAs have been studied as environmentally friendly replacement for petroleum-based polymers. High production costs and limited applications are impeding the widespread industrial adoption of PHAs, however. Genetic engineering of PHA-producing bacteria can help overcome this problem. Poly(hydroxyalkanoate) synthase (PHAS) is an enzyme responsible for PHA synthesis. In this presentation, we will discuss a study we undertake to genetically fuse the genes encoding the two subunits of a PHAS of Allochromatium vinosum ATCC 35206, i.e., the phaE and phaC genes. We hypothesize that the fused genes will facilitate the association of the two subunits to increase the product yield. Two fused genes (phaEC and phaCE) are constructed using the overlapping-primer-extension PCR method. The overlapping primers were designed in such a way that a stretch of 6 amino-acid sequence links the two fused pha genes in the resultant phaEC and phaCE genes. The progress of the ongoing experiments to express the fused genes in Pseudomonas resinovorans and Ralstonia eutropha using an expression vector, pBS29-P2, will be discussed in detail in the presentation. The results should yield important information for improving production yield of PHA in genetically modified organisms.