重组Kex2蛋白酶

Kex2蛋白酶是钙离子依赖性的蛋白水解酶，能特异性识别和切割Arg-Arg、Lys-Arg等双碱基氨基酸羧基端肽键，与胰蛋白酶不同，Kex2不能识别和切割单一碱性氨基酸即精氨酸或赖氨酸的羧基端肽键。在酵母体内Kex2蛋白酶负责加工killer toxin和α-factor的前体。Kex2蛋白酶是由毕赤酵母表达生产。酶的最适pH为9.0，最适温度为37℃。在偏酸性环境中稳定，稳定pH为 5.0-6.0。Kex2蛋白酶活性不受常规的丝氨酸蛋白酶抑制剂如PMSF、TPCK所抑制。 Kex2蛋白酶推荐使用方法： 如果溶解后不立即使用，推荐溶解缓冲：20mM NaAc-HAc, 2 mM Ca2+, pH 5.0~5.5. 推荐酶切缓冲液：5-10 mM Tris, 2mM Ca2+, pH 7.0-9.0 or HEPES, 5mM Ca2+, pH 7.0-9.0. 推荐条件： 蛋白酶：融合蛋白质量比1:20~1:1000 Kex2 is a Ca2+-dependent serine protease and cleaves at C-terminal site of Lys-Arg, Arg-Arg, Pro-Arg in pro-α,factor and killer-toxin precursors maturing, it was discovered in Saccharomyces cerevisiae.But Kex2 can’t recognize and cut a single basic amino acid,such as carboxyl end peptide bond of arginine and lysine. Recombinant Kex2 is a genetically engineered protein expressed in Pichia pastoris and purified by high pressure liquid chromatography. The optimal pH of Kex2 protease is 9.0, and the optimal temperature is 37 ℃. It is stable in buffer (pH 5.0-6.0). The activity of Kex2 is not affected by the conventional serine protease inhibitors such as PMSF, TPCK, TLCK inhibition. Kex2 Protease Recommended Method: Recommended reaction buffer: 5-10 mM Tris, 2mM Ca2+, pH 7.0-9.0 or HEPES, 5mM Ca2+, pH 7.0-9.0. If it is not used immediately after dissolution, it is recommended to use 20mM NaAc-HAc, 2mM Ca2+ (pH 5.0~5.5) buffer to dissolve the lyophilized powder. During the reaction, use 5-10 mM Tris, 2mM Ca2+, pH 7.0-9.0 or HEPES, 5mM Ca2+, pH 7.0-9.0 as the reaction diluent. Recommended conditions: Protease: Fusion protein Ratio 1:20~1:1000.

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