Isolation and heterologous expression of PHA synthesising genes from Bacillus thuringiensis R1

TitleIsolation and heterologous expression of PHA synthesising genes from Bacillus thuringiensis R1
Publication TypeJournal Article
Year of Publication2008
AuthorsDesetty, RD, Mahajan, VS, Khan, BMohammad, Rawal, SK
JournalWorld Journal of Microbiology & Biotechnology
Volume24
Issue9
Pagination1769-1774
Date PublishedSEP
Type of ArticleArticle
ISSN0959-3993
Keywordsaceto-acetyl-coA reductase, Bacillus thuringiensis R1, PHA synthase, polyhydroxyalkanoate
Abstract

The polyhydroxyalkanoate biosynthesis gene locus from Bacillus thuringiensis R1 was isolated, cloned and analyzed at the molecular level. We found that a similar to 5 kb SacI-ClaI digested fragment of genomic DNA from B. thuringiensis R1 encoding the PHA synthesising genes, conferred PHA producing ability to E. coli. The fragment was sequenced and found to be of 4787 bp with five open reading frames. Sequence alignment with closely related species of Bacillus in the existing database revealed that the ORFs correspond to phaP, phaQ, phaR, phaB and phaC genes. However, E. coli harboring phaP, phaQ, phaR, phaB and phaC locus produced very low PHA. Furthermore, complementation of the locus with phaA from Ralstonia eutropha increased the PHA production in the recombinant E. coli from 3.0% to 24% of cell dry mass. The putative promoter regions and ribosome binding sites were identified for each of the gene. Conserved domains for PHA synthase and aceto-acetyl-coA reductase were also identified. We hence conclude that the PHA operon of Bacillus thuringiensis R1 consists of phaP, phaQ, phaR, phaB, phaC and complementation of the same with phaA is accountable for its high PHA production.

DOI10.1007/s11274-008-9669-7
Type of Journal (Indian or Foreign)Foreign
Impact Factor (IF)1.532
Divison category: 
Biochemical Sciences