| 摘要 |
PURPOSE:To produce catechols in high efficiency, by culturing a catechol- producing microbial strain containing benzene oxygenase and benzene glycol dehydrogenase gene in a medium containing benzenes. CONSTITUTION:A chromosome DNA of Pseudomonas putida 98-136 (FERM P-6970) is prepared and the DNA is cut with a restriction enzyme Sau3A to obtain a DNA fragment of about 30kb. A plasmid vector pMMB 33 is cut separately with restriction enzymes SmaI and HpaI, the cut fragments are mixed, cut with BamHI, linked with the above DNA fragment of 30kb with a T4 ligase and introduced into E.coli to obtain the objective E.coli SI-300 (FERM BP-1358) capable of accumulating catechol from benzene. The SI-300 stain is cultured in a medium containing benzene, toluene, ethylbenzene, etc., and the objective catechols can be separated from cultured liquid. |
