利用Bacillus subtilis168脂肪酶LipA提高Sulfolobus shibatae B12嗜热酯酶B12est的可溶性

摘要

Esterase B12est from thermoacidophilic archaeon Sulfolobus shibatae B12 was cloned and over- expressed in E. coli as an inclusion body form, thereby,fusion the lipase LipA from Bacillus subtilis168 to the N terminal of B12est and the fused enzyme expressed as a soluble form. Results of this study showed that the fusion enzyme reserved the characterization of thennophilic esterase and lipase. The fused enzyme could catalyze p- nitrophenyl (p- NP) esters and triacylglycerides. Even in the high temperature, the fused enzyme still have the catalyze activity, it is highly thenrmostable and retain more than 60％ of its initial activity after incubation at 60℃ for 30 min.%克隆并在E.coli中表达嗜热古菌Sulfolobus shibatae B12中的B12est酯酶发现是以不可溶的形式表达,通过在其N端融合来源于Bacillus subtilis168中的脂肪酶LipA后,得到了具有一定可溶性的融合酶.本研究表明融合酶保留了嗜热酯酶和脂肪酶的酶学特性.融合酶能够催化对硝基苯酚酯和甘油三酯,在高温下仍然具有一定的催化活力,融合酶在60℃处理30 min后,还能够保持60%以上的活力.