Title Cloning and Expression in E. coli of the Genes Responsible for Degradation of 4-Chlorobenzoate and 4-Chlorocatechol drom Pseudomonas sp. Strain S-47
Author Kim, Ki Pil · Seo, Dong In · Lee, Dong Hun · Kim, Young Soo¹ · Kim, Chi Kyung *
Address Laboratory of Environmental and Molecular Microbiology, Division of Life Sciences, College of Natural Sciences; ¹Department of Pharmacy, Chungbuk National University
Bibliography Journal of Microbiology, 36(2),99-105, 1998,
Key Words benABCD, xylE 4-chlorobenzoate, 4-Chlorocatechol, dioxygenase, Pseudomonas sp. S-47
Abstract Pseudomonas sp. strain S-47 can grow on 4-chlorobenzoate (4CBA) and transform 4CBA to 4-chlorocatechol (4CC) under aerobic conditions, which is subsequently degraded to produce 2-hydroxypent-2, 4-dienoate (2H-2,4DA). The upper steps for conversion of 4CBA to 4CC are recognized to be conducted by the benzoate-1,2-dioxygenase (B12O) system encoded by benABC and benD. The ensving meta-cleabage reaction of 4CC is catalyzed by catechol 2,3-dioxygenase(C23O) encoded by the xylE gene. The benABCD and the xylE genes were cloned from the chromosome of Pseudomonas sP. S-47 into pCS1(48.7kb), pCS101(24.4kb), pCS201(17.7kb), and pCS202(6.7kb) recombinant plasmids, and were well ecpressed in E. coli XL1-Blue host cells. The PstI-insert (4.0kb) of pC202 was found to contain the benABCD and cylE genes and to have 2 EcoRV, 1 SphI, and 3 SacII restriction sites.
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