Poly-beta-hydroxybutyrate (PHB) biosynthetic genes in Rhizobium meliloti 41.
Genes encoding beta-ketothiolase (phaA), acetoacetyl-CoA reductase (phaB) and PHB-synthase (phaC) from R. meliloti 41, together with a fourth gene, referred to as ORF1, presumed to be involved in PHB biosynthesis, have been cloned and sequenced. phaA, phaB and ORF1 were identified by heterologous hybridization on a cosmid library, while phaC was isolated by cloning the transposon-tagged fragment from a R. meliloti PHB- Tn5 mutant. phaA and phaB were functionally expressed in Escherichia coli while phaC was able to complement a PHB- strain of R. meliloti 41. The three genes were sufficient to direct the production of polyhydroxyalkanoate in E. coli. The homology of ORF1 with an ORF located near the PHB genes in two phototrophic bacteria suggests its involvement in PHB synthesis.
Tombolini
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CRIBI Biotechnology Center, Università di Padova, Italy.
Povolo
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Buson
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Squartini
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Nuti
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D000102:Acetyl-CoA C-Acyltransferase / Q000235:genetics; D000217:Acyltransferases / Q000235:genetics; D000429:Alcohol Oxidoreductases / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006885:Hydroxybutyrates / Q000378:metabolism*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011091:Polyesters / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D016962:Sinorhizobium meliloti / Q000235:genetics*; D000094122:Polyhydroxybutyrates
D000102:Acetyl-CoA C-Acyltransferase; D000217:Acyltransferases; D000429:Alcohol Oxidoreductases; D006885:Hydroxybutyrates; D011091:Polyesters; D011994:Recombinant Proteins; C003182:poly-beta-hydroxybutyrate; D000094122:Polyhydroxybutyrates; C023744:acetoacetyl-CoA reductase; C060970:poly-beta-hydroxybutyrate polymerase