Expression of Cholesterol Oxidase with Temperature Induced Vector pHsh

doi:10.3969/j.issn.1673-1689.2016.03.008

Home > Archive>Volume 35, Issue 3, 2016 >272-277. DOI:10.3969/j.issn.1673-1689.2016.03.008

Expression of Cholesterol Oxidase with Temperature Induced Vector pHsh
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                        10.3969/j.issn.1673-1689.2016.03.008
                    
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Cholesterol oxidase is of great value to be applied in medicine，food and agriculture. In this study，cholesterol oxidase from Rhodococcus sp. was constructed into the temperature-induced vector pHsh and then by controlling the culture temperature，the gene was successfully expressed in E.coli without adding inducer. The fermentation optimization of cholesterol oxidase was carried out in the shake flask level. With the optimum condition，the enzyme activity reached 855 U/L. When purified to homogeneity using Ni-NTA，the resulting enzyme had the molecular mass of 55 kDa，the optimal pH 7.0 and temperature 40℃. The enzyme was also stable over a pH range of 5.0~7.0，with t1/2 of 6.9 min at 50 ℃. With cholesterol as substrate at pH 7.5 and 37 ℃，the kinetic parameters Km and kcat/Km were 3.38 mM and 6.09 s-1·mM-1，respectively.

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YUAN Ye, ZHANG Ling, SHAO Weilan, WANG Wu, YANG Hailin. Expression of Cholesterol Oxidase with Temperature Induced Vector pHsh[J]. Journal of Food Science and Biotechnology,2016,35(3):272-277.

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Online: November 01,2016
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