期刊
ACS SYNTHETIC BIOLOGY
卷 11, 期 1, 页码 486-496出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.1c00570
关键词
Trichoderma reesei; heterologous protein expression; chassis; iterative gene deletion
资金
- National Science and Technology Major Project of China [2018YFA0900500]
- National Natural Science Foundation of China [31921006, 31741003]
- Strategic Biological Resources Service Network Plan of the Chinese Academy of Sciences [KFJ-BRP-009]
- International Great Science Program of the Chinese Academy of Sciences [153D31KYSB20170121]
By employing a genome editing system and iterative gene deletion, marker-free T. reesei chassis were constructed, resulting in reduced secretion of native proteins and extracellular protease activity, leading to increased production levels of heterologous proteins.
Trichoderma reesei has an extremely high capacity for synthesizing and secreting proteins, thus exhibiting promise as an expression platform for heterologous proteins. However, T. reesei secretes large amounts of native proteins, which hinders its widespread application for heterologous protein production. Here, we designed and built a series of T. reesei chassis using an iterative gene deletion approach based on an efficient genome editing system. Donor DNAs with specially designed construct facilitated screening of positive deletion strains without ectopic insertion. Finally, marker-free T. reesei chassis with lower rates of native protein secretion and low levels of extracellular protease activity were constructed after 11 consecutive rounds of gene deletion. Higher production levels of three heterologous proteins-a bacterial xylanase XYL7, a fungal immunomodulatory protein LZ8, and the human serum albumin HSA-were achieved with these chassis using the cbh1 promoter. It is possible that diverse high-value proteins might be produced at a high yield using this engineered platform.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据