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Biochemistry (Moscow)

, Volume 74, Issue 5, pp 502–508 | Cite as

Enzymatic properties of a recombinant phospholipid hydroperoxide glutathione peroxidase from Momordica charantia and its complementation function in yeast

  • Chun-Juan Dong
  • Xiao-Dong Yang
  • Jin-Yuan LiuEmail author
Article

Abstract

The entire encoding region for Momordica charantia phospholipid hydroperoxide glutathione peroxidase (McPHGPx) was cloned into pET-28a(+) vector and expressed in Escherichia coli BL21(DE3). The purified recombinant McPHGPx displayed GSH-dependent peroxidase activity towards phospholipid hydroperoxide, H2O2, and tert-butyl hydroperoxide and had the highest affinity with and catalytic efficiency for phospholipid hydroperoxide. The optimum temperature of the enzyme activity ranged from 40 to 50°C, thus it is a thermostable enzyme compared to other PHGPx enzymes. Furthermore, McPHGPx expression in Saccharomyces cerevisiae PHGPx-deletion mutant rescued the susceptibilities to the oxidation-sensitive polyunsaturated fatty acid (linolenic acid), indicating its PHGPx complementation function in yeast. These results have well documented that McPHGPx functions as a PHGPx in vitro and in vivo and will be beneficial for further functional studies on plant PHGPx enzymes.

Key words

phospholipid hydroperoxide glutathione peroxidase Momordica charantia enzymatic properties functional complementation antioxidant enzyme 

Abbreviations

BHT

butylated hydroxytoluene

GPx

glutathione peroxidase

IPTG

isopropyl β-D-thiogalactopyranoside

LA

linolenic acid

(Mc)PHGPx

(Momordica charantia) phospholipid hydroperoxide glutathione peroxidase

MDA

malondialdehyde

PCOOH

phosphatidylcholine hydroperoxide

ROS

reactive oxygen species

SD/Trp

synthetic complete medium without tryptophan

TBA

thiobarbituric acid

t-BHP

tert-butyl hydroperoxide

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Copyright information

© Pleiades Publishing, Ltd. 2009

Authors and Affiliations

  1. 1.Laboratory of Molecular Biology and MOE Laboratory of Protein Science, Department of Biological Sciences and BiotechnologyTsinghua UniversityBeijingChina

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