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OALib Journal期刊
ISSN: 2333-9721
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痤疮丙酸杆菌亚油酸异构酶的克隆表达及酶学性质研究
DOI: 10.3969/j.issn.1000-2006.2014.06.020, PP. 105-109
Keywords: 痤疮丙酸杆菌,亚油酸异构酶,共轭亚油酸,酶学性质
Abstract:
为了研究使用生物酶法制备共轭亚油酸的可行性,笔者对痤疮丙酸杆菌亚油酸异构酶进行了异源表达和理化性质研究。根据大肠杆菌偏好密码子,优化了来源于痤疮丙酸杆菌亚油酸异构酶(pai)的基因(pai),将克隆的基因在bl21(de3)中进行表达,纯化目的蛋白,获得重组pai蛋白。sds-page结果显示,重组pai分子质量为48ku,重组蛋白主要以包涵体沉淀的形式存在。经测定重组pai在35℃、ph7.0时酶活最高,比酶活为752.3μmol/(min？mg)。经35℃保温2h,酶活保持90%以上,ph为6.5～7.0时,pai具有较好的稳定性,mn2+和zn2+对酶活力分别有22.4%和15.8%的抑制作用,以亚油酸为底物时该酶的km值为1.13mmol/l,kcat为4.67s-1。相关分析表明,痤疮丙酸杆菌亚油酸异构酶性质优良,适合以亚油酸为底物生物催化制备共轭亚油酸。
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