摘要
谷氨酰胺转氨酶(TGase或TG)是一种可催化蛋白质间形成异肽键,使蛋白质改性的天然酶制剂。本文以蛹虫草基因组为模板,通过PCR扩增得到TGase相似蛋白的基因片段tgM,将其与大肠杆菌-毕赤酵母穿梭表达载体pPIC9K连接,构建重组表达载体pPIC9K-TG,在毕赤酵母中进行表达,实现目的蛋白的胞外分泌表达,结果发酵液中重组TGase的酶活力为100 U/L。本研究为TGase的异源表达及潜在的工业应用提供参考。
Transglutaminase(TGase or TG),one of the natural food cross-linking agents with strong cross-linking function,form network structures among proteins viaε-(γ-glutamyl)-lysine(G-L)bonds.The gene tgM of TGase was obtained from the genomic DNA of Cordyceps militaris through PCR amplification.It was inserted into an E.coli-P.pastoris shuttle vector pPIC9K to construct recombinant plasmid pPIC9K-TG.The recombinant TGase was successfully expressed in P.pastoris and secreted into the culture medium.In optimized fermentation conditions,the activity of TGase in fermentation broth was up to 100 U/L.The results could provide guidance for the heterologous expression and potential industrial application of TGase.
作者
杨聪
郭丽琼
叶志伟
邹苑
余颖豪
林俊芳
Yang Cong;Guo Liqiong;Ye Zhiwei;Zou Yuan;Yu Yinghao;Lin Junfang(College of Food Science,South China Agricultural University,Guangzhou 510640;Research Center for Micro-Ecological Agent Engineering and Technology of Guangdong Province,Guangzhou 510640)
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2022年第2期107-113,共7页
Journal of Chinese Institute Of Food Science and Technology
基金
国家自然科学基金项目(31772373,32072646,31901693)。
关键词
谷氨酰胺转氨酶
毕赤酵母
异源表达
蛹虫草
transglutaminase
Pichia pastoris
heterologous expression
Cordyceps militaris
