Putative phospholipid hydroperoxide glutathione peroxidase from Antrodia camphorata

Antrodia camphorata

SpeciesNiu-Chang-Chih

JournalFood chemistry

Year2009

Key points

Glutathione Peroxidases (GPxs) play important roles in antioxidation
A cDNA (Ac-PHGPx, 764bp) encoding a putative phospholipid hydroperoxide glutathione peroxidase (PHGPx) from Antrodia camphorata has been cloned
To characterize the Ac-PHGPx, the coding region was subcloned into pYEX-S1 and transformed into Saccharomyces cerevisiae
The purified enzyme showed a predominant band with molecular mass of ~18kDa on 12% SDS-PAGE. The enzyme retained 50% activity at 60°C for 8min
The enzyme showed 42% activity after incubation with trypsin at 37°C for 40min
In addition, the ability of Ac-PHGPx to protect intact supercoiled plasmid DNA from OH induced nicking was demonstrated

From the paper

Abstract

Glutathione Peroxidases (GPxs) play important roles in antioxidation. A cDNA (Ac-PHGPx, 764bp) encoding a putative phospholipid hydroperoxide glutathione peroxidase (PHGPx) from Antrodia camphorata has been cloned. The deduced amino acid sequence is conserved among the reported GPxs. To characterize the Ac-PHGPx, the coding region was subcloned into pYEX-S1 and transformed into Saccharomyces cerevisiae. The recombinant 6His-tagged Ac-PHGPx was expressed and purified by Ni²⁺-nitrilotriacetic acid Sepharose. The purified enzyme showed a predominant band with molecular mass of ~18kDa on 12% SDS-PAGE. The enzyme retained 50% activity at 60°C for 8min. The enzyme was most active at pH 9. The enzyme showed 42% activity after incubation with trypsin at 37°C for 40min. In addition, the ability of Ac-PHGPx to protect intact supercoiled plasmid DNA from OH induced nicking was demonstrated.

Citation

Chen HT, Lin CY, Ken CF, Wen L, Lin CT (2009). Putative phospholipid hydroperoxide glutathione peroxidase from Antrodia camphorata. Food chemistry https://doi.org/10.1016/j.foodchem.2008.12.038

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