牛肉风味强化肽表达条件的初步研究  

Primary Study on Expression of Beefy Meaty Peptide in Recombinant Pichia Pastoris

作　　者：王艳萍[1] 张建[1] 杨永清[1] 

机构地区：[1]天津市食品营养与安全重点实验室天津科技大学食品工程与生物技术学院,天津300222

出　　处：《中国食品学报》2006年第1期230-234,共5页Journal of Chinese Institute Of Food Science and Technology

基　　金：天津市重大科技攻关项目(No.05ZHGCNC00200)

摘　　要：牛肉风味强化肽(BeefyMeatyPeptide,BMP)最初是从牛肉的消化液中分离的八肽,能增强鲜味,可作为一种类似于谷氨酸钠的天然风味强化剂。文章介绍了利用基因工程技术在酵母菌中表达BMP的方法,对构建的Pichiapastoris酵母工程菌株X-33/pPICZαA-BMP进行了发酵表达,确定了菌株的表型、高密度生长及表达条件。试验结果显示:构建的工程菌株为甲醇利用慢表型Muts,优化后的种子培养基摇瓶条件下菌体密度OD600达到28。在pH3.0、诱导72h、甲醇添加量为0.5%的条件下BMP有最大表达,选用BSM有利于诱导表达BMP;建立了SephadexG-25分离纯化、RP-HPLC检测BMP的有效方法。Beefy meaty peptide （BMP） is an octapeptide with delicious taste, first isolated from beef digested with papain. There were synergistic effects of salt and MSG on BMP and BMP can be used as a natural flavor enhancer like MSG. This paper introduced the expression of BMP in Pichia Pastoris using recombinant DNA technology, determined the phenotype of X-33/ pPICZαA-BMP, optimized the growth conditions and expression conditions. The results showed that the recombinant strain was Mut^s, OD600 reached 28 at optimized conditions, and BMP has the largest expression at pH 3.0, inducing 72 h. We selected BSM as expression medium, and BMP was purified by Sephadex G-25 and then detected by RP-HPLC.

关 键 词：牛肉风味肽 毕赤酵母 表达 

分 类 号：TS251.52[轻工技术与工程—农产品加工及贮藏工程]