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KMID : 0043319920150040360
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Archives of Pharmacal Research
1992 Volume.15 No. 4 p.360 ~ p.364
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Cloning and Overexpression of Methylcatechol 2,3-Dioxygenase Gene from Toluene-Degrading Pseudomonas putida mt-2 (pWWO)
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Lee JR
Min KR/Kim YS
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Abstract
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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.
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KEYWORD
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