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酵母乙醇提取物对重组大肠杆菌外源蛋白表达的影响
朱岩岩,熊爱英,桑秀梅,王晶,王期,殷志敏
0
(南京师范大学生命科学学院,生物化学与生物制品研究所,江苏省分子医学重点实验室,南京 210046)
摘要:
酵母乙醇提取物是利用啤酒干酵母经培养发酵后提取的富含多种活性物质的混合物。初步实验研究发现,该物质对大肠杆菌蛋白表达有明显促进作用。为了进一步研究其对蛋白表达的作用,以E.coliBL21(DE3)/pGEX-4t-1-ggt为主要研究菌,以2 g·L-1乳糖作为诱导剂,研究酵母乙醇提取物对γ-GGT蛋白表达的作用。研究发现,分批添加终浓度10 g·L-1的酵母提纯提取物到2 g·L-1的乳糖为诱导剂的培养基中可以显著提高γ-GGT蛋白的表达及宿主菌的生长,蛋白表达量比1 mmol·L-1 IPTG诱导效果高10%~50%,菌体密度提高1.75~3倍左右。对于其他的重组蛋白及表达宿主有相似的作用。酵母乙醇提取物与乳糖组成的复合制剂可以有效替代有毒性的IPTG作为诱导剂的使用,为大规模诱导蛋白表达奠定了基础。
关键词:  酵母乙醇提取物  乳糖  诱导  蛋白合成
DOI:10.13610/j.cnki.1672-352x.20160512.029
投稿时间:2016-01-19
基金项目:江苏省科技厅产学研联合创新资金-前瞻性研究项目(BY2013001-03)资助。
Effect of Yeast extract on heterologous protein production in recombinant Escherichia coli
ZHU Yanyan,XIONG Aiying,SANG Xiumei,WANG Jing,WANG Qi,YIN Zhimin
(School of life Sciences ,Nanjing Normal University, Institute of Biochemistry and Biological Products, Jiangsu Province Key Laboratory for Molecular and Medical Biotechnology, Nanjing 210046)
Abstract:
The ethanol extract of yeast contains a variety of active substances extracted from beer yeast. Preliminary study found that the ethanol extract can significantly promote protein expression in E. coli. Our laboratory used E.coliBL21 (DE3) / pGEX-4t-1-ggt as the main research bacteria and added 2 g·L-1 lactose to the medium as an inducer to study the role of yeast alcohol extract on protein expression of γ-GGT .We found that γ-GGT protein expression yield and the growth of the host bacteria can be significantly improved when the final concentration at 10 g·L-1 of yeast extract was batch added into medium containing 2 g·L-1 of lactose at different fermentation periods. The results showed that the amount of expressed protein increased by 10%~50% comparing with that induced by 1 mmol·L-1 IPTG and cell density increased by 1.75-3 times. Similar effect has been shown for different proteins expressed in recombinant Escherichia coli. The mixture of ethanol yeast extract and lactose may be the effective alternative inducer to replace toxic IPTG. This result would lay a foundation for large-scale protein expression in the future.
Key words:  yeast extract  lactose  induce  protein synthesis

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