Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4

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 weigh...

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Main Authors: Kong, Christina Wen Hui, Tan, Irene Kit Ping, Zazali Alias
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2022
Online Access:http://journalarticle.ukm.my/21464/
http://journalarticle.ukm.my/21464/1/M%2025.pdf
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author Kong, Christina Wen Hui
Tan, Irene Kit Ping
Zazali Alias,
author_facet Kong, Christina Wen Hui
Tan, Irene Kit Ping
Zazali Alias,
author_sort Kong, Christina Wen Hui
building UKM Institutional Repository
collection Online Access
description 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 PputUW4_00801 (putative glutathione S-transferase) of Pseudomonas sp. UW4 according to its estimated molecular weight and the pI values obtained experimentally.
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spelling oai:generic.eprints.org:214642023-04-07T00:54:46Z http://journalarticle.ukm.my/21464/ Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4 Kong, Christina Wen Hui Tan, Irene Kit Ping Zazali Alias, 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 PputUW4_00801 (putative glutathione S-transferase) of Pseudomonas sp. UW4 according to its estimated molecular weight and the pI values obtained experimentally. Penerbit Universiti Kebangsaan Malaysia 2022 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/21464/1/M%2025.pdf Kong, Christina Wen Hui and Tan, Irene Kit Ping and Zazali Alias, (2022) Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4. Malaysian Applied Biology, 51 (4). pp. 177-184. ISSN 0126-8643 https://jms.mabjournal.com/index.php/mab/index
spellingShingle Kong, Christina Wen Hui
Tan, Irene Kit Ping
Zazali Alias,
Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4
title Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4
title_full Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4
title_fullStr Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4
title_full_unstemmed Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4
title_short Bioinformatic analysis and purification of Glutathione Transferase (GST) from Pseudomonas sp. UW4
title_sort bioinformatic analysis and purification of glutathione transferase (gst) from pseudomonas sp. uw4
url http://journalarticle.ukm.my/21464/
http://journalarticle.ukm.my/21464/
http://journalarticle.ukm.my/21464/1/M%2025.pdf