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. 2005 Nov;187(21):7543-5.
doi: 10.1128/JB.187.21.7543-7545.2005.

Replacement of tyrosine 181 by phenylalanine in gentisate 1,2-dioxygenase I from Pseudomonas alcaligenes NCIMB 9867 enhances catalytic activities

Chew Ling Tan  1 , Chew Chieng Yeo, Hoon Eng Khoo, Chit Laa Poh
Affiliations

Replacement of tyrosine 181 by phenylalanine in gentisate 1,2-dioxygenase I from Pseudomonas alcaligenes NCIMB 9867 enhances catalytic activities

Chew Ling Tan et al. J Bacteriol. 2005 Nov.

Abstract

xlnE, encoding gentisate 1,2-dioxygenase (EC 1.13.11.4), from Pseudomonas alcaligenes (P25X) was mutagenized by site-directed mutagenesis. The mutant enzyme, Y181F, demonstrated 4-, 3-, 6-, and 16-fold increases in relative activity towards gentisate and 3-fluoro-, 4-methyl-, and 3-methylgentisate, respectively. The specific mutation conferred a 13-fold higher catalytic efficiency (k(cat)/Km) on Y181F towards 3-methylgentisate than that of the wild-type enzyme.

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References

    1. Andujar, E., and E. Santero. 2003. Site-directed mutagenesis of an extradiol dioxygenase involved in tetralin biodegradation identifies residues important for activity or substrate specificity. Microbiology 149:1559-1567. - PubMed
    1. Bradford, M. M. 1976. A rapid and sensitive method for quantification of microgram quantities of proteins utilizing the principle of protein dye binding. Anal. Biochem. 72:248-254. - PubMed
    1. Crawford, R. L., S. W. Hutton, and P. J. Chapman. 1975. Purification and properties of GDO from Moraxella osloensis. J. Bacteriol. 121:794-799. - PMC - PubMed
    1. Feng, Y., H. E. Khoo, and C. L. Poh. 1999. Purification and characterization of gentisate 1,2-dioxygenases from Pseudomonas alcaligenes NCIB 9867 and Pseudomonas putida NCIB 9869. Appl. Environ. Microbiol. 65:946-950. - PMC - PubMed
    1. Harpel, M. R., and J. D. Lipscomb. 1990. Gentisate 1,2-dioxygenase from Pseudomonas. Substrate coordination to active site Fe2+ and mechanism of turnover. J. Biol. Chem. 265:22187-22196. - PubMed

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