Molecular cloning of structural and regulatory hydrogenase (hox) genes of Alcaligenes eutrophus H16

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The Alcaligenes eutrophus H16 hoxX gene participates in hydrogenase regulation.

Nucleotide sequence analysis revealed a 1,791-bp open reading frame in the hox gene cluster of the gram-negative chemolithotroph Alcaligenes eutrophus H16. In order to investigate the biological role of this open reading frame, we generated an in-frame deletion allele via a gene replacement strategy. The resulting mutant grew significantly more slowly than the wild type under lithoautotrophic c...

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Identification and molecular characterization of the Alcaligenes eutrophus H16 aco operon genes involved in acetoin catabolism.

Acetoin:dichlorophenolindophenol oxidoreductase (Ao:DCPIP OR) and the fast-migrating protein (FMP) were purified to homogeneity from crude extracts of acetoin-grown cells of Alcaligenes eutrophus. Ao:DCPIP OR consisted of alpha and beta subunits (Mrs, 35,500 and 36,000, respectively), and a tetrameric alpha 2 beta 2 structure was most likely for the native protein. The molecular weight of FMP s...

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Role of hydrogen in the activation and regulation of hydrogen oxidation by the soluble hydrogenase from Alcaligenes eutrophus H16.

The activation kinetics of the H2-oxidizing activity of the soluble hydrogenase from Alcaligenes eutrophus H16 were investigated. Activation with Na2S2O4 plus 101 kPa H2 resulted in a rapid increase in activity over 1 h and constant activity after 3 h incubation. Less-stable activations were achieved if enzyme was incubated with Na2S2O4 under 1 kPa H2 or 101 kPa N2. The enzyme could also be par...

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Nickel requirement for active hydrogenase formation in Alcaligenes eutrophus.

The nickel-dependent chemolithoautotrophic growth of Alcaligenes eutrophus is apparently due to a requirement of nickel for active hydrogenase formation. Cells grown heterotrophically with fructose and glycerol revealed a specific activity of soluble and membrane-bound hydrogenase which was severalfold higher than the normal autotrophic level. The omission of nickel from the medium did not affe...

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Cloning and expression of the levanase gene in Alcaligenes eutrophus H16 enables the strain to hydrolyze sucrose

Genetic engineering methods were used to enhance the substrate spectrum of Alcaligenes eutrophus H16, a poly-fl-hydroxybutyric acid (PHB) producer. Using parts of the vector pMMB33 and a 2.5 kb DNA fragment of the Bacillus subtilis chromosome a plasmid was constructed bearing the gene for levanase, an enzyme able to hydrolyze various saccharides. After transfer of the levanase gene by triparent...

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ژورنال

عنوان ژورنال: Journal of Bacteriology

سال: 1986

ISSN: 0021-9193,1098-5530

DOI: 10.1128/jb.168.2.636-641.1986