Archives of Pharmacal Research
- Volume 15 Issue 4
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- Pages.360-364
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- 1992
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- 0253-6269(pISSN)
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- 1976-3786(eISSN)
Cloning and Overexpression of Methylcatechol 2, 3-Dioxygenase Gene from Toluene-Degrading Pseudomonas putida mt-2(pWWO)
- Lee, Jeong-Rai (Department of Biochemistry, College of Pharmacy, Chungbuk National University) ;
-
Min, Kyung-Rak
(Department of Biochemistry, College of Pharmacy, Chungbuk National University)
;
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Kim, Young-Soo
(Department of Biochemistry, College of Pharmacy, Chungbuk National University)
- Published : 1992.12.01
Abstract
Methylcatechol 2, 3-dioxygenase encoded in pWWO megaplasmid of Pseudomonas putida mt-2 has been cloned and overexpressed in Escherichia coli. This enzyme gene has been localized inside 2. 3-kb XhoI fragment derived from the pWWO megaplasmid. Analysis of enzyme activity and SDS-PAGE showed that the cloned methylcatechol 2, 3-dioxygenase gene in E. coli was about 100 fold overexpressed compared with the parental gene in P. putida mt-2 (pWWO). The cloned enzyme exhibited higher ring-fission activity to catechol than catechol derivatives including 3-methylcatechol, 4-methylcatechol, and 4-chlorocatechol.