粘质沙雷氏菌低温脂肪酶的基因克隆与酶学性质分析

发布时间：2018-06-27 09:02

  本文选题：沙雷氏菌 + 脂肪酶　； 参考：《福州大学学报(自然科学版)》2016年05期

【摘要】：克隆一个源于北极冻土沙雷氏菌的脂肪酶基因lip18,实现其在大肠杆菌中的表达,并进行酶学性质研究.克隆脂肪酶基因lip18,并构建p ET-28a(+)-lip18重组表达载体,导入大肠杆菌BL21(DE3)中,诱导优化重组蛋白表达,并研究其酶学性质.克隆得到脂肪酶基因lip18,全长为1 842 bp,编码614个氨基酸.重组菌的诱导温度对蛋白表达影响很大,在最适诱导温度为20℃时脂肪酶大量表达,重组脂肪酶的相对分子质量约为65 ku.酶学性质研究表明:重组酶高效水解C10~C16的中、长链脂肪酸,最适作用温度30℃,最适作用pH值7.0,并且在0℃条件下有一定的催化活性,热稳定性差,是低温中性脂肪酶;同时,对有机溶剂有较好的耐受性.
[Abstract]:Lipase gene lip18 was cloned from Shareh's bacteria in the Arctic, and its expression in Escherichia coli was achieved and its enzymatic properties were studied. Lipase gene lip18 was cloned and pET-28a () -lip18 recombinant expression vector was constructed. The recombinant protein was transformed into E. coli BL21 (DE3), and its enzymatic properties were studied. Lip18 gene was cloned and its length was 1 842 BP, encoding 614 amino acids. The induction temperature of recombinant bacteria had a great influence on the protein expression, and the relative molecular weight of the recombinant lipase was about 65 ku. at the optimal induction temperature of 20 鈩，

本文编号：2073392