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Identification of PHA loci in Thermus thermophilus HB8 genome

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Identification of PHA loci in Thermus thermophilus HB8 genome
Pantazaki, A. A.
Kyriakidis, D.A.
Mimikakou, G. E.
Polyhydroxyalkanoic acids (PHAs) are carbon and energy storage polymers synthesized by most eubacteria and archea in response to environmental conditions. The PHA biosynthetic bacteria are classified into four groups based on the organization of the pha gene locus and the structure-function properties of the PHA synthases. Synthases are the key enzymes in the biosynthesis of PHA and catalyse the stereoselective conversion of (R)-3-hydroxyacyl-CoA substrates to PHAs. Recently we have published the purification and characterization of a PHA synthase from Thermus thermophilus HB8 [1]. Although the entire genome of T. thermophilus has been published [2] a large number of ORFs still remains as hypothetical proteins. In this study PCR-based methods were employed for the identification of pha biosynthesis genes in T. thermophilus genome. For pha loci amplification the LA Taq polymerase was used and genomic DNA isolated from the bacterial cells. The primers were designed based on the highly conserved nucleic acid sequences in regions of the class II pha loci of various Pseudomonas. PCR products were sucloned into PCRII and later into pUC19. Escherichia coli JM109 were transformed with the constructed plasmids and screened for PHA synthase activity. Three putative regions were identified and studied further. The amplification pattern indicates that T. thermophilus HB8 belongs to class II pha loci. Elucidation of the mechanism of PHA biosynthesis in a thermophilic microorganism may lead to polymers with improved properties of high biotechnological value
Article
National Hellenic Research Foundation (NHRF)
Helios - Repository of the National Hellenic Research Foundation (NHRF)
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English
2008
2012-06-12T12:23:07Z
2008-12-11T09:22:00Z
http://hdl.handle.net/10442/7932
DOI: http://dx.doi.org/10.1111/j.1742-4658.2008.06448.x
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