工业微生物2017，Vol.47Issue(4)：30-36,7.DOI:10.3969/j.issn.1001-6678.2017.04.005
Burkholderia cepacia胆固醇酯酶及其分子伴侣基因的串联表达及酶学性质的研究
Tandem expression of Burkholderia cepacia cholesterol esterase gene and its molecular chaperone
摘要
Abstract
In order to achieve the soluble expression of cholesterol esterase,Burkholderia cepacia ST-200 cholesterol esterase was tandem expressed in E.coli by adding SD sequence between the enzyme and its molecular chaperone.Analysis of SDS polyacrylamide gel electrophoresis showed that the soluble expression of cholesterol esterase realized successfully and the total amount of enzyme was 1 077 U/L in shake-flask fermentation.When purified to homogeneity using Ni-NTA,the resulting enzyme had a relative molecular mass of 37 kDa.Secondary structure of cholesterol esterase was analyzed using circular dichroism and the Tm value was measured.The optimum temperature,the optimum pH,thermal stability,pH stability and organic solvent tolerance of cholesterol esterase were studied and it indicated that cholesterol esterase had maximum activity at pH 7.0 and 45 ℃.关键词
胆固醇酯酶/Burkholderiacepacia/分子伴侣/串联表达Key words
cholesterol esterase/Burkholderia cepacia/molecular chaperone/tandem expression引用本文复制引用
张亚慧,张玲,沈微,辛瑜,杨海麟..Burkholderia cepacia胆固醇酯酶及其分子伴侣基因的串联表达及酶学性质的研究[J].工业微生物,2017,47(4):30-36,7.基金项目
江苏省优青(BK20160053) （BK20160053）
江苏省产学研(BY2016022-40). （BY2016022-40）
