Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1

Journal of Basic Microbiology
Azzmer Azzar Abdul HamidFahrul Huyop

Abstract

The non-stereospecific α-haloalkanoic acid dehalogenase DehE from Rhizobium sp. RC1 catalyzes the removal of the halide from α-haloalkanoic acid D,L-stereoisomers and, by doing so, converts them into hydroxyalkanoic acid L,D-stereoisomers, respectively. DehE has been extensively studied to determine its potential to act as a bioremediation agent, but its structure/function relationship has not been characterized. For this study, we explored the functional relevance of several putative active-site amino acids by site-specific mutagenesis. Ten active-site residues were mutated individually, and the dehalogenase activity of each of the 10 resulting mutants in soluble cell lysates against D- and L-2-chloropropionic acid was assessed. Interestingly, the mutants W34→A,F37→A, and S188→A had diminished activity, suggesting that these residues are functionally relevant. Notably, the D189→N mutant had no activity, which strongly implies that it is a catalytically important residue. Given our data, we propose a dehalogenation mechanism for DehE, which is the same as that suggested for other non-stereospecific α-haloalkanoic acid dehalogenases. To the best of our knowledge, this is the first report detailing a functional aspect for DehE, a...Continue Reading

References

Jan 1, 1991·Methods in Enzymology·J A Wells
Mar 21, 2008·Journal of Molecular Biology·Jason W SchmidbergerMatthew C J Wilce
Apr 8, 2010·Natural Product Reports·David O'Hagan, Jason W Schmidberger
Feb 11, 2011·Bioscience, Biotechnology, and Biochemistry·Tatsuo Kurihara

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Citations

May 8, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Thiau-Fu AngThean Chor Leow
Jun 28, 2016·SpringerPlus·Aliyu AdamuFahrul Huyop
Oct 24, 2020·Journal of Biomolecular Structure & Dynamics·Habeebat Adekilekun OyewusiRoswanira Abdul Wahab
May 21, 2021·Journal of Biomolecular Structure & Dynamics·Habeebat Adekilekun OyewusiAzzmer Azzar Abdul Hamid
Dec 3, 2021·Journal of Biomolecular Structure & Dynamics·Habeebat Adekilekun OyewusiFahrul Huyop

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