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Cloning and characterization of poly(3‐hydroxybutyrate) biosynthesis genes from Pseudomonas sp. USM 4‐55
Author(s) -
Tan Yifen,
Neo PeiChin,
Najimudin Nazalan,
Sudesh Kumar,
Muhammad Tengku Sifzizul Tengku,
Othman Ahmad Sofiman,
Samian Razip
Publication year - 2010
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.200900138
Subject(s) - operon , polyhydroxyalkanoates , biosynthesis , escherichia coli , pseudomonas , cupriavidus necator , biochemistry , heterologous expression , chemistry , atp synthase , biology , microbiology and biotechnology , gene , bacteria , recombinant dna , genetics
Pseudomonas sp. USM 4‐55 is a locally isolated bacterium that possesses the ability to produce polyhydroxyalkanoates (PHA) consisting of both poly(3‐hydroxybutyrate) [P(3HB)] homopolymer and medium‐chain length (mcl) monomers (6 to 14 carbon atoms) when sugars or fatty acids are utilized as the sole carbon source. In this study, the P(3HB) biosynthesis operon carrying the phbC Ps P(3HB) synthase was successfully cloned and sequenced using a homologous probe. Three open reading frames encoding NADPH‐dependent acetoacetyl‐coenzyme A reductase (PhbB Ps ), β‐ketothiolase (PhbA Ps ) and P(3HB) synthase (PhbC Ps ) were found in the phb operon. The genetic organization of phb operon showed a putative promoter region, followed by phbB Ps ‐ phbA Ps ‐ phbC Ps . phbR Ps which encoded a putative transcriptional activator was located in the opposite orientation, upstream of phbBAC Ps . Heterologous expression of pGEM″ABex harboring phbC Ps in Escherichia coli JM109 resulted in P(3HB) accumulation of up to 40% of dry cell weight (DCW). (© 2010 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)