microbiology, Vol 141, 2553-2559, Copyright © 1995 by Society for General Microbiology

ARTICLES

Poly-beta-hydroxybutyrate (PHB) biosynthetic genes in Rhizobium meliloti 41

R Tombolini, S Povolo, A Buson, A Squartini and MP Nuti
CRIBI Biotechnology Center, Universita di Padova, Italy.

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.

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