Cloning, Expression and Purification of Glutathione Peroxidase of Antarctic Yeast Rhodotorula mucilaginosa AN5

Hai Yu, Cuijuan Shi, Xiujun Gao, Guangfeng Kan

2022

Abstract

Glutathione peroxidase (GPx) is a key enzyme in glutathione antioxidant enzyme system, which can remove H2O2 and other oxidants in organisms. In this study, the GPx gene was cloned, sequence analyzed, prokaryotic expressed and purified from Antarctic yeast Rhodotorula mucilaginosa AN5. The GPx gene sequence was amplified successfully by PCR and named RmGPx (GenBank No. KX164292). The open reading frame (ORF) of RmGPx was 498 bp and encoded 165 amino acids. The predicted molecular weight of the protein was 18268.6 Da and the theoretical isoelectric point was 8.37. The recombinant expression plasmid pET-28a-RmGPx was successfully constructed and transferred to E. coli. The protein expression was the optimum when induced by 0.2 mM IPTG at 37 ℃ for 4 hours. The protein was purified by an elution buffer of 100 mM imidazole in Ni NTA column. All study results supply the theoretical foundation for the functional analysis and application of GPx protein.

Paper Citation

in Harvard Style

Yu H., Shi C., Gao X. and Kan G. (2022). Cloning, Expression and Purification of Glutathione Peroxidase of Antarctic Yeast Rhodotorula mucilaginosa AN5. In Proceedings of the 4th International Conference on Biomedical Engineering and Bioinformatics - Volume 1: ICBEB, ISBN 978-989-758-595-1, pages 553-557. DOI: 10.5220/0011232500003443

in Bibtex Style

@conference{icbeb22,
author={Hai Yu and Cuijuan Shi and Xiujun Gao and Guangfeng Kan},
title={Cloning, Expression and Purification of Glutathione Peroxidase of Antarctic Yeast Rhodotorula mucilaginosa AN5},
booktitle={Proceedings of the 4th International Conference on Biomedical Engineering and Bioinformatics - Volume 1: ICBEB,},
year={2022},
pages={553-557},
publisher={SciTePress},
organization={INSTICC},
doi={10.5220/0011232500003443},
isbn={978-989-758-595-1},
}

in EndNote Style

TY - CONF

JO - Proceedings of the 4th International Conference on Biomedical Engineering and Bioinformatics - Volume 1: ICBEB,
TI - Cloning, Expression and Purification of Glutathione Peroxidase of Antarctic Yeast Rhodotorula mucilaginosa AN5
SN - 978-989-758-595-1
AU - Yu H.
AU - Shi C.
AU - Gao X.
AU - Kan G.
PY - 2022
SP - 553
EP - 557
DO - 10.5220/0011232500003443