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TOL pismid size of Flavobacterium odoratum SUB53 was estimated as 83 Md and the optimum concentration of m-toluate degradation by TOL plasmid was 5 mM. H2 production by Rhodopseudomonas sphaeroides KCTC1425 was largely dependent on nitrogenase activity and showed the highest at 30 mM malate with 7 mM glutamate as nitrogen source. Nitrogenase activities were inhibited by 03 mM NH,´ ions, to be appeared the decrease of H, production. Conjugation of TOL plasmids from F. odoratum SUB53 and Pseudomonas putida mt-2 to R sphaeroides showed the optimum at the exponential stage of recipient cells in presence of helper plasmid pRK2013. According to the investigation of catechol-l,2-oxygenase (C-1, 20) and catechol-2,3-oxygenase (C-2,3-0) activities of R sphaeroides Cl (TOL SUB53) and C2 (TOL mt-2), the gene for C-2,3-0 is located on TOL plasmid and gene for C-1, 2-0 on the chromosome of R sphaeroides. m-Toluate was biodegraded by TOL plasmid in R sphaeroides Cl and C2, presumably to be produced H2 gas from the ? secondary metabolites of m-toluate.
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