Volume 4, Issue 1, February 2016, Page: 8-12
A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings
Md. Motiar Rohman, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Afsana Hoque Akhi, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Nusrat Jahan Methela, Department of Genetics and Plant Breeding, Patuakhali Science and Technology University, Patuakhali, Bangladesh
Mohammad Golam Hossain, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
M. Shalim Uddin, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Mohammad Amiruzzaman, Molecular Breeding Lab, Plant Breeding Division, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Bhagya Rani Banik, Training and Communication Wing, Bangladesh Agricultural Research Institute, Gazipur, Bangladesh
Received: Dec. 27, 2015;       Accepted: Jan. 8, 2016;       Published: Jan. 21, 2016
DOI: 10.11648/j.jps.20160401.12      View  3806      Downloads  110
Abstract
In this study an attempt was taken to purify Glyoxalase-I (Gly-I: E.C., 4.4.1.5), from maize seedlings. Both green and non-green parts of 7 day old maize seedlings were used as plant materials. Crude proteins were precipitated by 65% (NH4)2SO4, and dialyzed overnight. The dialyzate was applied on DEAE-cellulose chromatography and eluted with linear gradient of KCl from 0 to 0.2 M. In both cases, Gly-I eluted at approximately 85 mM of KCL. The active Gly-I fractions were pooled and applied on a hydroxylapatite chromatography and eluted with 0-40 mM potassium-phosphate buffer, but the eluted fractions showed very poor activity. Therefore, the active pooled fraction of DEAE-chromatography was then applied directly on affinity chromatography (S-hexyl glutathione-agarose) for final purification and eluted with 1.2 mM of S-hexyl glutathione. The purified protein from green and non-green part had specific activity of 33.23 and 39.25 μmol min-1 mg-1 protein, respectively, along with recovery of 1.47 and 162, respectively, and yield of 83.11 and 68.15, respectively. In SDS-PAGE, the active purified affinity fraction was found to move with another protein. The spectrophotometric analysis of high active Gly-I fractions from DEAE-cellulose and affinity chromatography showed GST [another detoxifying enzyme (E.C., 2.5.1.18)] activity. This result suggested that one of the adjacent protein bands in SDS-PAGE was due to presence of a GST in Gly-I fraction.
Keywords
Glyoxalase-I Purification, Glutathione S-transferase, Simultaneous Elution, Maize
To cite this article
Md. Motiar Rohman, Afsana Hoque Akhi, Nusrat Jahan Methela, Mohammad Golam Hossain, M. Shalim Uddin, Mohammad Amiruzzaman, Bhagya Rani Banik, A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings, Journal of Plant Sciences. Vol. 4, No. 1, 2016, pp. 8-12. doi: 10.11648/j.jps.20160401.12
Copyright
Copyright © 2016 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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