乳酸乳球菌表面表达Brazzein甜味蛋白系统的建立  

Surface Display of Brazzein in Lactococcus lactis

作　　者：王长远[1] 许凤[1] 沈冰蕾[1] 于长青[1] 姚笛[1] 

机构地区：[1]黑龙江八一农垦大学食品学院,大庆163319

出　　处：《天然产物研究与开发》2014年第8期1170-1173,共4页Natural Product Research and Development

基　　金：黑龙江省自然基金项目(QC2011C028)

摘　　要：参照甜味蛋白Brazzein基因序列,结合乳酸乳球菌的密码子偏嗜性的相关分析,对甜味蛋白Brazzein基因进行改造,将第29位天冬氨酸、31位的组氨酸和第41位的谷氨酸分别突变为赖氨酸、丙氨酸和赖氨酸,以期提高目的蛋白的甜度。采用重叠PCR的方法合成目的基因,目的片段亚克隆到pMD18-T载体上。经序列测定分析后,目的片段克隆入分泌表达载体pNZ8112,电转化乳酸乳球菌中,构建成表面展示表达甜味蛋白Brazzein的乳酸乳球菌表达系统。The gene encoding of Brazzein was modified according to the codon bios in Lactococcus lactls and three sense mutations in Brazzein, Asp29Lys, His31Ala, Glu41Lys were generated in an attempt to enhance the sweetness. The optimized gene was amplified by overlap PCR and the fragment was subcloned into pMD18-T vector,followed by being sequenced. The gene of interest was cloned into pNZ8112 expression vector and recombinant plasmid was transformed into Llactls by electroporation to construct the surface display system for expression of the Brazzein protein on the surface of L. hwtls.

关 键 词：甜味蛋白 BRAZZEIN 乳酸乳球菌 表面展示 

分 类 号：TS201.2[轻工技术与工程—食品科学]