Please use this identifier to cite or link to this item: https://doi.org/10.1016/S0378-1097(01)00393-7
Title: Identification of amino acid residues essential for catalytic activity of gentisate 1,2-dioxygenase from Pseudomonas alcaligenes NCIB 9867
Authors: Chua, C.H.
Feng, Y.
Yeo, C.C. 
Poh, C.L. 
Khoo, H.E. 
Keywords: Catalytic activity
Gentisate 1,2-dioxygenase
Mutagenesis
Issue Date: 2001
Source: 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 1.13.11.4) 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
URI: http://scholarbank.nus.edu.sg/handle/10635/31468
ISSN: 03781097
DOI: 10.1016/S0378-1097(01)00393-7
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

12
checked on Dec 13, 2017

WEB OF SCIENCETM
Citations

12
checked on Dec 13, 2017

Page view(s)

169
checked on Dec 9, 2017

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.