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题名: Biochemical characterization of 1-Cys peroxiredoxin from Antrodia camphorata
作者: Lisa Wen;Hui-Ming Huang;Rong-Huay Juang;Chi-Tsai Lin
贡献者: NTOU:Institute of Bioscience and Biotechnology
关键词: Mushroom;Antrodia camphorata;Expression;1-Cys peroxiredoxin (1-Cys Prx)
日期: 2007-01-01
上传时间: 2011-10-21T02:22:24Z
出版者: Applied Microbiology and Biotechnology
摘要: Abstract:Antrodia camphorata is a unique medicinal mushroom found only in Taiwan. It has been used as a remedy for various diseases in folk medicine. Antrodia camphorata has been shown to exhibit antioxidative effects. Peroxiredoxins play important roles in antioxidation and cell signaling. A gene encoding an antioxidant enzyme, 1-cysteine peroxiredoxin (1-Cys Prx), was identified in an expressed sequence tag database of the A. camphorata and cloned by polymerase chain reaction. The 1-Cys Prx cDNA (837 bp, accession no. AY870325) contains an open reading frame encoding a protein of 223 amino acid residues with calculated molecular mass of 25,081 Da. The deduced protein shared 44–58% identity with 1-Cys Prx from Homo sapiens, Bos taurus, and Saccharomyces cerevisia. The sequence surrounding the conserved cysteine DFTPVCTTE is conserved. The coding sequence was subcloned into a vector, pET-20b (+), and transformed into Escherichia coli. The recombinant 1-Cys Prx was purified by Ni2+-nitrilotriacetic acid (Sepharose). The purified enzyme was characterized under various conditions. The enzyme is thermostable because its half-life of inactivation was 15.5 min at 60°C. It was stable under alkaline pH range from 7.8 to 10.2. The enzyme showed decreased activity with increasing concentration of imidazole. The enzyme is sensitive to trypsin and chymotrypsin treatment.
關聯: 73(6), pp.1314-1322
显示于类别:[生命科學暨生物科技學系] 期刊論文


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