Bacillus megaterium, plasmid

Bacillus subtilis transformants were identified by their resistance to naphthalene, parachlorobiphenyl, or dibenzofuran. Controls subtilis protoplasts regenerated without the addition of B. megaterium plasmid DNA) exhibited no increased reistance to these three compounds. That each transformant was B. subtilis and not a contaminant was verified by testing for the B. subtilis chromosomal markers. Only streptomycin resistant strains that showed arginine and leucine dependent growth were used in subsequent experiments. 

The degradation of alkanoic acids by P-oxidation has been noted parenthetically above, but alternative pathways may occur. For example, the metabolism of hexanoic acid by strains of Pseudomonas sp. may take place by co-oxidation with subsequent formation of succinate and 2-tetrahydrofurany-lacetate as a terminal metabolite (Kunz and Weimer 1983). In a strain of Cory neb acterium sp., the specificities of the relevant catabolic enzymes are consistent with the production of dodecanedioic acid by co-oxidation of dode-cane but not of hexadecanedioic acid from hexadecane (Broadway et al. 1993). Hydroxylation at subterminal (co-1, co-2, and co-3) positions of carboxylic acids with chain lengths of 12 to 18—and less readily of the corresponding alcohols, but not the carboxylic acids or the alkanes—has been observed (Miura and Fulco 1975) for a soluble enzyme system from a strain of Bacillus megaterium. Whereas in this organism co-2 hydroxylation is carried out by a soluble cytochrome P-450 BM 3 (Narhi and Fulco 1987), co-hydroxylation in P. oleovorans that carries the OCT plasmid is mediated by a three-component hydroxylase that behaves like a cytoplasmic membrane protein (Ruettinger et al. 1974 Kok et al. 1989). 

BacUhis megaterium to Bacillus subtiBs by Plasmid Transfer Techniques... 

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