Identification of a lipoamide dehydrogenase gene as second locus affected in poly(3-hydroxybutyric acid)-leaky mutants of Alcaligenes eutrophus.
From a genomic library of Alcaligenes eutrophus strain H16 in the broad-host range cosmid pVK100 a 6300-bp EcoRI-fragment was cloned, which restored the wild-type phenotype in transposon-induced poly(3-hydroxybutyric acid)-leaky mutants, derived from A. eutrophus. Nucleotide sequence analysis of the region adjacent to the transposon insertion revealed an open reading frame which complied with all criteria for a coding region. This region was referred to as phbL, and the deduced amino acid sequence from this part of phbL showed 60% amino acid identity in an overlap of 98 residues to the lipoamide dehydrogenase gene (lpd) of Escherichia coli. In addition, the 6300-bp EcoRI-fragment conferred expression of lipoamide dehydrogenase activity to E. coli.
Pries
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Institut für Mikrobiologie, Georg-August-Universität Göttingen, FRG.
Hein
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Steinbüchel
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D020155:3-Hydroxybutyric Acid; D000421:Alcaligenes / Q000145:classification / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics*; D008058:Dihydrolipoamide Dehydrogenase / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D006885:Hydroxybutyrates / Q000378:metabolism*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D017422:Sequence Analysis, DNA; D014162:Transfection / Q000235:genetics*
D004269:DNA, Bacterial; D006885:Hydroxybutyrates; D008058:Dihydrolipoamide Dehydrogenase; D020155:3-Hydroxybutyric Acid