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题名: Taiwanofungus Camphorata Nitroreductase: cDNA Cloning and Biochemical Characterisation
作者: Chen, Chih-Chen;Ken, Chuian-Fu;Wen, Lisa;Chang, Ching-Fong;Lin, Chi-Tsai
贡献者: 生物學系
关键词: Nitroreductase;Taiwanofungus camphorata;Three-dimension structural model (3-D structural model)
日期: 2012-12
上传时间: 2013-06-05T08:48:29Z
出版者: Elsevier
摘要: Nitroreductases (Nrs) play important roles in redox system via NADPH or NADH as a reductant. A TcNr cDNA encoding a putative Nr was cloned from Taiwanofungus camphorata. A 3-D structural model of the TcNr has been created based on the known structure of BcNr (Bacillus cereus). To characterise the TcNr, the coding region was subcloned into an expression vector and transformed into Escherichia coli. The recombinant His6-tagged TcNr was purified by Ni affinity chromatography. The purified enzyme showed a single band at molecular mass of approximately 25 kDa on 12% sodium dodecyl sulphate–polyacrylamide gel electrophoresis. The enzyme exhibited Nr activity via ferricyanide assay. The Michaelis constant (KM) value for ferricyanide was 0.86 mM. The enzyme’s half-life of deactivation at 45 °C was 12.3 min. The enzyme was most active at pH 6. The enzyme’s preferred substrate is 1-chloro-2, 4-dinitrobenzene.
關聯: Food Chemistry, 135(4): 2708-2713
显示于类别:[生物學系] 期刊論文


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