BIOINFORMATIC ANALYSIS AND PURIFICATION OF GLUTATHIONE TRANSFERASE (GST) from Pseudomonas sp. UW4
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Published:2022-10-31
Issue:4
Volume:51
Page:177-184
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ISSN:2462-151X
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Container-title:Malaysian Applied Biology
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language:
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Short-container-title:MAB
Author:
KONG CHRISTINA WEN HUI,TAN IRENE KIT PING,ALIAS ZAZALI
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 PputUW4_00801 (putative glutathione S-transferase) of Pseudomonas sp. UW4 according to its estimated molecular weight and the pI values obtained experimentally.
Publisher
Persatuan Biologi Gunaan Malaysia
Subject
General Agricultural and Biological Sciences
Reference23 articles.
1. Allocati, N., Casalone, E., Masulli, M., Ceccarelli, I., Carletti, E., Parker, M. W. & Di Ilio, C. 1999. Functional analysis of the evolutionarily conserved proline 53 residue in Proteus mirabilis glutathione transferase B1-1. FEBS Letters, 445: 347–350. 2. Allocati, N., Federici, L., Masulli, M. & Di Ilio, C. 2009. Glutathione transferases in bacteria, FEBS Journal, 276: 58–75. 3. Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72:248-54. 4. Bresell, A., Weinder R., Lundqvist, G., Raza, W., Shimoji, M., Sun, T-H., Balk, L., Wiklund, R., Erikson, J., Jansson, C., Persson, B., Jakobsson, P-J. & Morgentenson, R. 2005. Bioinformatic and enzymativ characterization of the MAPEG superfamily. FEBS Journal, 272: 1688-1703. 5. Di Ilio, C., Aceto, D.A., Piccolomini, R., Allocati, N., Faraone, A., Cellini, L., Ravagnani, G. & Federici, G. 2001. Purification and characterization of three forms of glutathione transferase from Proteus mirabilia. Biochemical Journal, 360:1–16.
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