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.
| Published in | Journal of Plant Sciences (Volume 4, Issue 1) |
| DOI | 10.11648/j.jps.20160401.12 |
| Page(s) | 8-12 |
| Creative Commons |
This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited. |
| Copyright |
Copyright © The Author(s), 2024. Published by Science Publishing Group |
Glyoxalase-I Purification, Glutathione S-transferase, Simultaneous Elution, Maize
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APA Style
Md. Motiar Rohman, Afsana Hoque Akhi, Nusrat Jahan Methela, Mohammad Golam Hossain, M. Shalim Uddin, et al. (2016). 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, 4(1), 8-12. https://doi.org/10.11648/j.jps.20160401.12
ACS Style
Md. Motiar Rohman; Afsana Hoque Akhi; Nusrat Jahan Methela; Mohammad Golam Hossain; M. Shalim Uddin, et al. A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings. J. Plant Sci. 2016, 4(1), 8-12. doi: 10.11648/j.jps.20160401.12
AMA Style
Md. Motiar Rohman, Afsana Hoque Akhi, Nusrat Jahan Methela, Mohammad Golam Hossain, M. Shalim Uddin, et al. A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings. J Plant Sci. 2016;4(1):8-12. doi: 10.11648/j.jps.20160401.12
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Download@article{10.11648/j.jps.20160401.12,
author = {Md. Motiar Rohman and Afsana Hoque Akhi and Nusrat Jahan Methela and Mohammad Golam Hossain and M. Shalim Uddin and Mohammad Amiruzzaman and Bhagya Rani Banik},
title = {A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings},
journal = {Journal of Plant Sciences},
volume = {4},
number = {1},
pages = {8-12},
doi = {10.11648/j.jps.20160401.12},
url = {https://doi.org/10.11648/j.jps.20160401.12},
eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.jps.20160401.12},
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.},
year = {2016}
}
TY - JOUR T1 - A Glutathione S-transferase Elutes with Glyoxalase-I (Gly-I) During Purification of Gly-I from Maize (Zea mays L.) Seedlings AU - Md. Motiar Rohman AU - Afsana Hoque Akhi AU - Nusrat Jahan Methela AU - Mohammad Golam Hossain AU - M. Shalim Uddin AU - Mohammad Amiruzzaman AU - Bhagya Rani Banik Y1 - 2016/01/21 PY - 2016 N1 - https://doi.org/10.11648/j.jps.20160401.12 DO - 10.11648/j.jps.20160401.12 T2 - Journal of Plant Sciences JF - Journal of Plant Sciences JO - Journal of Plant Sciences SP - 8 EP - 12 PB - Science Publishing Group SN - 2331-0731 UR - https://doi.org/10.11648/j.jps.20160401.12 AB - 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. VL - 4 IS - 1 ER -
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