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OALib Journal期刊
ISSN: 2333-9721
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定点突变对扩展青霉脂肪酶热稳定性的改善
DOI: 10.3724/SP.J.1145.2013.00043, PP. 43-47
Keywords: 扩展青霉,脂肪酶,定点突变,毕赤酵母,热稳定性
Abstract:
利用重叠延伸pcr方法对扩展青霉脂肪酶基因进行体外定点突变，构建了k115r与随机突变体ep8双突变的重组质粒pao815-ep8-k115r.将该质粒电转化到毕赤酵母gs115中进行异源表达，获得双突变体脂肪酶pel-ep8-k115r-gs.与野生型脂肪酶pel-gs和随机突变体脂肪酶pel-ep8-gs进行比较，发现双突变体脂肪酶pel-ep8-k115r-gs在40℃温浴30min后残余酶活为89%，比野生型脂肪酶pel-gs和随机突变体脂肪酶pel-ep8-gs分别提高54%和27%；双突变体脂肪酶pel-ep8-k115r-gs的tm值为42.2℃，比野生型脂肪酶pel-gs提高3.5℃，比随机突变脂肪酶pel-ep8-gs提高2.0℃.研究结果表明对扩展青霉脂肪酶的分子改造能提高其热稳定性.
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