一种用于在里氏木霉工业菌株RUT-C30中进行异源蛋白共表达和纯化的新型宿主-载体系统。
A novel host-vector system for heterologous protein co-expression and purification in the Trichoderma reesei industrial strain RUT-C30.
作者信息
Zhang Lei, Zhang Suping, Jiang Xin, Wei Wei, Wang Wei, Wei Dongzhi
机构信息
State Key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Meilong Road, P.O.B. 311, 130, Shanghai, 200237, China.
Research Center for Biomass Energy, East China University of Science and Technology, Shanghai, 200237, China.
出版信息
Biotechnol Lett. 2016 Jan;38(1):89-96. doi: 10.1007/s10529-015-1948-4. Epub 2015 Sep 5.
OBJECTIVES
To develop a host-vector system for heterologous protein co-expression and purification in the Trichoderma reesei (teleomorph Hypocrea jecorina) industrial strain RUT-C30.
RESULTS
The co-expression and purification system is based on: (i) an efficient and reliable selectable marker using bar (phosphinothricin acetyltransferase gene); (ii) a compact hygromycin B resistance marker; and (iii) a versatile integration plasmid for gene expression and knockout. Fluorescent protein genes were introduced into T. reesei and the corresponding proteins were purified from fermentation broth. The host-vector system was used in a proof-of-principle approach to achieve the co-expression of an alkaline endoglucanase and an alkaline cellobiohydrolase.
CONCLUSIONS
This protocol can be used at an industrial scale to produce large amounts of proteins in T. reesei.
目的
开发一种宿主-载体系统，用于在里氏木霉（有性型为杰氏肉座菌）工业菌株RUT-C30中进行异源蛋白的共表达和纯化。
结果
该共表达和纯化系统基于：（i）使用bar（膦丝菌素乙酰转移酶基因）的高效可靠的选择标记；（ii）紧凑的潮霉素B抗性标记；以及（iii）用于基因表达和敲除的通用整合质粒。将荧光蛋白基因导入里氏木霉，并从发酵液中纯化相应的蛋白质。该宿主-载体系统用于原理验证方法，以实现碱性内切葡聚糖酶和碱性纤维二糖水解酶的共表达。
结论
该方案可用于工业规模，在里氏木霉中大量生产蛋白质。
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