Bioinformatic analysis and purification of glutathione transferase (GST) from Pseudomonas sp. UW4
Document Type
Article
Publication Date
1-1-2022
Abstract
The study aimed at identifying and purifying cytosolic glutathione transferase isoforms expressed in Pseudomonas sp. UW4. Search at UniProt (https://www.uniprot.org/uniprot/), has indicated that there were 20 genes encoding putative glutathione transferases for the microorganism. The molecular weights of the isoforms ranged from 17.6 to 34.06 kDa. SDS-polyacrylamide gel electrophoresis revealed that the GST purified using Sulfobromophthalein-glutathione (BSP) affinity column, resolved into a single band with a low molecular weight (MW) of 16 kDa with the pI value of 6.0. Purified GST was reactive towards ethacrynic acid, 1-chloro-2,4-dinitrobenzene, cumene hydroxide, and hydrogen peroxide, but no detectable activity with Trans-2-octenal, hepta-2,4-dienal and Trans-4-phenyl-3-butene-2-one. This has proven that putative GST possessed peroxidase activity and proposed to be similar to PputUW4₀₀₈₀₁ (putative glutathione S-transferase) of Pseudomonas sp. UW4 according to its estimated molecular weight and the pI values obtained experimentally. © 2022, Malaysian Society of Applied Biology. All rights reserved.
Keywords
Affinity chromatography, glutathione transferases, Pseudomonas sp. UW4
Divisions
InstituteofBiologicalSciences
Funders
Kementerian Sains, Teknologi dan Inovasi [Grant no. FP034-2008C],Universiti Malaya [Grant no. PG176-2015A]
Publication Title
Malaysian Applied Biology
Volume
51
Issue
4