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Functional Role of Tyr12 in the Catalytic Activity of Novel Zeta-like Glutathione S-transferase from Acidovorax sp. KKS102

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Abstract

Glutathione S-transferases (GSTs) are a family of enzymes that function in the detoxification of variety of electrophilic substrates. In the present work, we report a novel zeta-like GST (designated as KKSG9) from the biphenyl/polychlorobiphenyl degrading organism Acidovorax sp. KKS102. KKSG9 possessed low sequence similarity but similar biochemical properties to zeta class GSTs. Functional analysis showed that the enzyme exhibits wider substrate specificity compared to most zeta class GSTs by reacting with 1-chloro-2,4-dinitrobenzene (CDNB), p-nitrobenzyl chloride (NBC), ethacrynic acid (EA), hydrogen peroxide, and cumene hydroperoxide. The enzyme also displayed dehalogenation function against dichloroacetate, permethrin, and dieldrin. The functional role of Tyr12 was also investigated by site-directed mutagenesis. The mutant (Y12C) displayed low catalytic activity and dehalogenation function against all the substrates when compared with the wild type. Kinetic analysis using NBC and GSH as substrates showed that the mutant (Y12C) displayed a higher affinity for NBC when compared with the wild type, however, no significant change in GSH affinity was observed. These findings suggest that the presence of tyrosine residue in the motif might represent an evolutionary trend toward improving the catalytic activity of the enzyme. The enzyme as well could be useful in the bioremediation of various types of organochlorine pollutants.

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Abbreviations

GST:

Glutathione S-transferase

CDNB:

1-Chloro-2,4-dinitrobenzene

NBC:

p-Nitrobenzyl chloride

EA:

Ethacrynic acid

CuOOH:

Cumene hydroperoxide

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Acknowledgements

We wish to thank Dr. Yuji Nagata of the graduate school of life sciences, Tohoku University, Japan, for granting us the permission to use the strain. We also wish to thank Japan collection of microorganism (JCM) for providing us with the strain. This work was supported by the University of Malaya IPPP [PG170-2016A]. One of the authors (DS) would like to thank Bayero University, Nigeria, for the financial assistance.

Funding

This study was funded by University of Malaya IPPP Grant [PG170-2016A].

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Authors and Affiliations

  1. Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603, Kuala Lumpur, Malaysia

    Dayyabu Shehu & Zazali Alias

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  1. Dayyabu Shehu
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  2. Zazali Alias
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Correspondence to Zazali Alias.

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Shehu, D., Alias, Z. Functional Role of Tyr12 in the Catalytic Activity of Novel Zeta-like Glutathione S-transferase from Acidovorax sp. KKS102. Protein J 37, 261–269 (2018). https://doi.org/10.1007/s10930-018-9774-x

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