Abstract
We identified the poly(hydroxyalkanoate) synthase (PHAS) genes of three strains of Pseudomonas oleovorans by using polymerase chain reaction (PCR)-based detection methods. P. oleovorans NRRL B-14682 contains Class I PHA synthase gene (phaC), NRRL B-14683 harbors Class II phaC1 and phaC2 genes, and NRRL B-778 contain both the Class I and II PHA synthase genes. Inverse-PCR and chromosomal walking techniques were employed to obtain the complete sequences of the Class I phaCs of NRRL B-778 (phbC778; 1698 bps) and B-14682 (phbC14682; 1899 bps). BLAST search indicated that these genes are new and had not been previously cloned. The gene product of phbC778 (i.e., PhbC778; 566 amino acid residues) is homologous to the Class I PHA synthases of Pseudomonas sp. HJ-2 and Pseudomonas sp. strain 61-3, and that of phbC14682 (PhbC14682; 632 amino acids) is homologous to PHAS of Delftia acidovorans. The PhbC14682 contains an extra sequence of 33 amino acids in its conserved α/β-hydrolase domain, making it only the second Class I PHA synthase found to contain this cellular proteolytic sequence. Consistent with their Pseudomonas origin, the codon-usage profiles of PhbC778 and PhbC14682 are similar to those of Pseudomonas Class II PHASs. These new Pseudomonas Class I phbC genes provide valuable addition to the gene pool for the construction of novel PHASs through gene shuffling.
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Literature Cited
IS Aldor JD Keasling (2003) ArticleTitleProcess design for microbial plastic factories: metabolic engineering of polyhydroxyalkanoates Curr Opin Biotechnol 14 475–483 Occurrence Handle1:CAS:528:DC%2BD3sXot12iu70%3D Occurrence Handle14580576
SF Altschul TL Madden AA Schäffer J Zhang Z Zhang W Miller DJ Lipman (1997) ArticleTitleGapped BLAST and PSI-BLAST: a new generation of protein database search programs Nucleic Acids Res 25 3389–3402 Occurrence Handle10.1093/nar/25.17.3389 Occurrence Handle1:CAS:528:DyaK2sXlvFyhu7w%3D Occurrence Handle9254694
RD Ashby DKY Solaiman TA Foglia (2002a) ArticleTitleThe synthesis of short- and medium-chain-length poly(hydroxyalkanoate) mixtures from glucose- or alkanoic acid-grown Pseudomonas oleovorans J Ind Microbiol Biotechnol 28 147–153 Occurrence Handle1:CAS:528:DC%2BD38XitV2nt7g%3D
RD Ashby DKY Solaiman TA Foglia (2002b) ArticleTitlePoly(ethylene glycol)-mediated molar mass control of short-chain- and medium-chain-length poly(hydroxyalkanoates) from Pseudomonas oleovorans Appl Microbiol Biotechnol 60 154–159 Occurrence Handle1:CAS:528:DC%2BD38XnvVOqur8%3D
P Besse H Veschambre (1994) ArticleTitleChemical and biological synthesis of chiral epoxides Tetrahedron 50 8885–8927 Occurrence Handle1:CAS:528:DyaK2cXmtlSmu7Y%3D
R Chenna H Sugawara T Koike R Lopez TJ Gibson DG Higgins JD Thompson (2003) ArticleTitleMultiple sequence alignment with the Clustal series of programs Nucleic Acids Res 31 3497–3500 Occurrence Handle1:CAS:528:DC%2BD3sXltVWjsrg%3D Occurrence Handle12824352
W Hazenberg B Witholt (1997) ArticleTitleEfficient production of medium-chain-length poly(3-hydroxyalkanoates) from octane by Pseudomonas oleovorans: economic considerations Appl Microbiol Biotechnol 48 588–596 Occurrence Handle1:CAS:528:DyaK2sXnsl2ku7s%3D
GW Huisman E Wonink R Meima B Kazemier P Terpstra B Witholt (1991) ArticleTitleMetabolism of poly(3-hydroxyalkanoates) (PHAs) by Pseudomonas oleovorans. Identification and sequences of genes and function of the encoded proteins in the synthesis and degradation of PHA J Biol Chem 266 2191–2198 Occurrence Handle1:CAS:528:DyaK3MXmt1Cqs7k%3D Occurrence Handle1989978
BHA Rehm (2003) ArticleTitlePolyester synthases: natural catalysts for plastics Biochem J 376 15–33 Occurrence Handle1:CAS:528:DC%2BD3sXovVajs7c%3D Occurrence Handle12954080
J Sambrook DW Russell (2001) Molecular cloning: a laboratory manual, 3rd ed, vol. 2 Cold Spring Harbor Laboratory Press Cold Spring Harbor, NY 881–885
D-S Sheu Y-T Wang C-Y Lee (2000) ArticleTitleRapid detection of polyhydroxyalkanoate-accumulating bacteria isolated from the environment by colony PCR Microbiology 146 2019–2025 Occurrence Handle1:CAS:528:DC%2BD3cXmtVemtrw%3D Occurrence Handle10931906
DKY Solaiman (2002) ArticleTitlePolymerase-chain-reaction-based detection of individual polyhydroxyalkanoate synthase phaC1 and phaC2 genes Biotechnol Lett 24 245–250 Occurrence Handle1:CAS:528:DC%2BD38Xitl2gsbs%3D
DKY Solaiman (2003) ArticleTitleBiosynthesis of medium-chain-length poly(hydroxyalkanoates) with altered composition by mutant hybrid PHA synthases J Ind Microbiol Biotechnol 30 322–326 Occurrence Handle1:CAS:528:DC%2BD3sXltFCntbw%3D Occurrence Handle12761653
DKY Solaiman RD Ashby TA Foglia (2000) ArticleTitleRapid and specific identification of medium-chain-length polyhydroxyalkanoate synthase gene by polymerase chain reaction Appl Microbiol Biotechnol 53 690–694 Occurrence Handle1:CAS:528:DC%2BD3cXkvVyhurc%3D Occurrence Handle10919328
A Steinbüchel T Lütke-Eversloh (2003) ArticleTitleMetabolic engineering and pathway construction for biotechnological production of relevant polyhydroxyalkanoates in microorganisms Biochem Eng J 16 81–96
K Sudesh T Fukui Y Doi (1998) ArticleTitleGenetic analysis of Comamonas acidovorans polyhydroxyalkanoate synthase and factors affecting the incorporation of 4-hydroxybutyrate monomer Appl Environ Microbiol 64 3437–3443 Occurrence Handle1:CAS:528:DyaK1cXmtVajtL8%3D Occurrence Handle9726894
TA Tatusova TL Madden (1999) ArticleTitleBlast 2 sequences - a new tool for comparing protein and nucleotide sequences FEMS Microbiol Lett 174 247–250 Occurrence Handle1:CAS:528:DyaK1MXjtlOlu74%3D Occurrence Handle10339815
T Tsuge S-i Imazua K Takaseb S Taguchib Y Doi (2004) ArticleTitleAn extra large insertion in the polyhydroxyalkanoate synthase from Delftia acidovorans DS-17: its deletion effects and relation to cellular proteolysis FEMS Microbiol Lett 231 77–83 Occurrence Handle1:CAS:528:DC%2BD2cXhtFWnsr4%3D Occurrence Handle14769470
JB Beilen ParticleVan G Eggink H Enequist R Bos B Witholt (1992) ArticleTitleDNA sequence determination and functional characterization of the OCT-plasmid-encoded alkJKL genes of Pseudomonas oleovorans Mol Microbiol 6 3121–3136 Occurrence Handle1453953
JB Beilen Particlevan S Panke S Lucchini AG Franchini M Röthlisberger B Witholt (2001) ArticleTitleAnalysis of Pseudomonas putida alkane-degradation gene clusters and flanking insertion sequences: evolution and regulation of the alk genes Microbiology 147 1621–1630 Occurrence Handle11390693
B Witholt B Kessler (1999) ArticleTitlePerspectives of medium chain length poly(hydroxyalkanoates), a versatile set of bacterial bioplastics Curr Opin Biotechnol 10 279–285 Occurrence Handle1:CAS:528:DyaK1MXjvVSnsLw%3D Occurrence Handle10361079
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The technical assistance of Nicole Cross and Marshall Reed is acknowledged.
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Solaiman, D.K., Ashby, R.D. Genetic Characterization of the Poly(hydroxyalkanoate) Synthases of Various Pseudomonas oleovorans Strains. Curr Microbiol 50, 329–333 (2005). https://doi.org/10.1007/s00284-005-4508-7
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DOI: https://doi.org/10.1007/s00284-005-4508-7