甜味蛋白Brazzein在乳酸乳球菌的表面表达及鉴定  

Surface Expression and Identification of Brazzein Exposed on the Outer Wall of Lactococcus lactis

作　　者：王长远[1] 刘祎婷 张丽萍[1] 姚笛[1] 

机构地区：[1]黑龙江八一农垦大学食品学院,黑龙江大庆163319 [2]齐齐哈尔疾病预防防控中心,黑龙江齐齐哈尔161005

出　　处：《中国食品学报》2015年第5期189-193,共5页Journal of Chinese Institute Of Food Science and Technology

基　　金：黑龙江省自然科学基金项目(QC2011C028)

摘　　要：为了提高甜味蛋白Brazzein的产量及甜度,将第29位的天冬氨酸、第31位的组氨酸和第41位的谷氨酸突变为赖氨酸、丙氨酸、赖氨酸,然后结合乳酸乳球菌密码子偏嗜性,对甜味蛋白编码序列进行优化。将优化的Brazzein基因克隆入表面表达载体p NZ8112,构建甜味蛋白Brazzein的乳酸乳球菌表面表达系统。通过SDSPAGE、免疫印迹及免疫荧光等方法验证甜味蛋白Brazzein成功表达于乳酸乳球菌表面,这为甜味蛋白Brazzein的商品化开发奠定基础。In an attempt to commercial utilization of Brazzein, the three sense mutations in Brazzein, Asp29Lys, His31Ala, Glu41Lys was generated and coding sequence of Brazzein gene was optimized according to the codon bios in Lactococcus lactis to enhance the sweetness and improve the yield. The optimized gene was cloned into pNZS112 sur- face-expression vector and recombinant plasmid was transformed into L. lactis to construct the surface-expression system for expression of the Brazzein protein on the surface of L. lactis. The results demonstrated the surface display system was successfully constructed upon the verification of SDS-PAGE, western blot and immunostaining analysis. The completion of surface- expression of Brazzein present on L. lactis indicates the great potential of commercial utilization.

关 键 词：甜味蛋白 BRAZZEIN 乳酸菌 表面表达 

分 类 号：TS201.3[轻工技术与工程—食品科学]