Please use this identifier to cite or link to this item:
|Title:||Identification of amino acid residues essential for catalytic activity of gentisate 1,2-dioxygenase from Pseudomonas alcaligenes NCIB 9867|
|Citation:||Chua, C.H., Feng, Y., Yeo, C.C., Poh, C.L., Khoo, H.E. (2001). Identification of amino acid residues essential for catalytic activity of gentisate 1,2-dioxygenase from Pseudomonas alcaligenes NCIB 9867. FEMS Microbiology Letters 204 (1) : 141-146. ScholarBank@NUS Repository. https://doi.org/10.1016/S0378-1097(01)00393-7|
|Abstract:||Gentisate 1,2-dioxygenase (GDO, EC 220.127.116.11) is a ring cleavage enzyme that utilizes gentisate as a substrate yielding maleylpyruvate as the ring fission product. Mutant GDOs were generated by both random mutagenesis and site-directed mutagenesis of the gene cloned from Pseudomonas alcaligenes NCIB 9867. Alignment of known GDO sequences indicated the presence of a conserved central core region. Mutations generated within this central core resulted in the complete loss of enzyme activity whereas mutations in the flanking regions yielded GDOs with enzyme activities that were reduced by up to 78%. Site-directed mutagenesis was also performed on a pair of highly conserved HRH and HXH motifs found within this core region. Conversion of these His residues to Asp resulted in the complete loss of catalytic activity. Mutagenesis within the core region could have affected quaternary structure formation as well as cofactor binding. A mutant enzyme with increased catalytic activities was also characterized. © 2001 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.|
|Source Title:||FEMS Microbiology Letters|
|Appears in Collections:||Staff Publications|
Show full item record
Files in This Item:
There are no files associated with this item.
checked on May 19, 2018
WEB OF SCIENCETM
checked on Apr 3, 2018
checked on May 12, 2018
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.