Characterization of Recombinant Cosmids Containing H_2-Oxidation Genes from Alcaligenes hydrogenophilus

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Ten Pseudomonas oxalaticus OX1 harboring recombinant cosmids containing H_2-oxidation genes (hox) from Alcaligenes hydrogenophilus showed H_2-dependent autotrophic growth with different growth rates. Some strains grew autotrophically almost as fast as the strain containing the Hox plasmid pHG21-a. Ten strains had high activity of membrane-bound hydrogenase (MH), but the activity of soluble hydrogenase could not be detected. Sizes of DNA inserts in recombinant cosmids isolated from 10 strains were 26 to 29 kilobases (Kb), and there was a 22-Kb overlapped region. It is suggested that the DNA fragment required for conferring the abillity of H_2-dependent autotrophic growth on CO_2-fixing P. oxalaticus appears to span a region of more than 20Kb in A. hydrogenophilus. Structural genes of MH were found in the right end of the overlapped region by DNA hybridization analysis using oligonucleotide probes corresponding to the conserved sequences of MH genes in Bradyrhizobium japonicum and Rhodobacter capsulatus. The MH was induced by H_2 and repressed by the addition of fructose, as in A. hydrogenophilus. Therefore, the inserted DNA fragment contained the DNA region which determined the regulatory properties as well as structural genes of MH.
社団法人日本生物工学会の論文
1989-10-25