产气荚膜梭菌磷脂酶C的重组表达及其脱胶应用.pdfVIP

66 CHINA OILS AND FATS摇 摇 摇 摇 摇 摇 摇 摇 摇 摇 摇 摇 摇 摇 2014 Vol郾39 No郾5 综合利用 产气荚膜梭菌磷脂酶C的重组表达及其脱胶应用 芮丽莲,余榛榛,梁摇 丽,常摇 明,金青哲,王兴国 (江南大学食品学院,食品科学与技术国家重点实验室,食品安全与营养协同创新中心,江苏无锡214122) 摘要:将产气荚膜梭菌(Clostridiumperfringens)磷脂酶C(Cp-PLC)基因在大肠杆菌BL21(DE3)中 进行了重组表达,并对重组Cp-PLC表达条件进行了优化。 结果表明:当菌体生长至OD 为1郾0 600 时,添加8 g/ L乳糖在30益下诱导32 h,得到的重组Cp-PLC 的酶活最大,为398.35 U/ mL;重组 Cp -PLC 用于菜籽毛油脱胶的最佳条件为温度57益、pH5.0、加酶量600 U/ kg、反应时间1.5 h;在 最佳条件下,菜籽毛油的磷含量可降至5.66 mg/ kg,能够满足物理精炼的要求。 关键词:磷脂酶C;产气荚膜梭菌;重组表达;脱胶;菜籽毛油 中图分类号:TS224.6;Q936摇 摇 文献标志码:A摇 摇 文章编号:1003-7969(2014)05-0066-05 Recombinant expression of Clostridiumperfringens phospholipase C and its application in degumming RUI Lilian,YU Zhenzhen,LIANG Li,CHANG Ming, JIN Qingzhe,WANG Xingguo (Synergetic Innovation Center of Food Safety and Nutrition,State Key Laboratory of Food Science and Technology,School of Food Science and Technology,Jiangnan University, Wuxi214122,Jiangsu, China) Abstract:The phospholipase C gene from Clostridium perfringens was recombinationally expressed in E.coli BL21(DE3),andtheexpressionconditionsofrecombinant Clostridiumperfringens phospholipaseC (Cp-PLC)were optimized.Theresultsshowedthatundertheconditionsof absorbanceof cellsat600 nm 1.0,lactose dosage8g/ L,inductiontemperature30益 andinductiontime32 h,theactivity ofrecombi鄄 nant Cp-PLCobtainedreached398.35 U/ mL. Theoptimal degumming conditionsof cruderapeseedoil with the recombinant Cp -PLC were obtained as follows:temperature 57益,pH 5.0,enzyme dosage 600 U/ kg,reaction time 1.5 h. Under these conditions,the phosphorus content of degummed rapeseed oil decreased to5.66 mg/ kg,which could fulfill the requirem