Lee Hyang-Mi, Ren Jun, Tran Kha Mong, Jeon Byeong-Min, Park Won-Ung, Kim Hyunjoo, Lee Kyung Eun, Oh Yuna, Choi Myungback, Kim Dae-Sung, Na Dokyun
Department of Biomedical Engineering, Chung-Ang University, Seoul, Republic of Korea.
Department of Biotechnology, Korea University, Seoul, Republic of Korea.
Commun Biol. 2021 Feb 15;4(1):205. doi: 10.1038/s42003-021-01726-w.
In bacterial biotechnology, instead of producing functional proteins from plasmids, it is often necessary to deliver functional proteins directly into live cells for genetic manipulation or physiological modification. We constructed a library of cell-penetrating peptides (CPPs) capable of delivering protein cargo into bacteria and developed an efficient delivery method for CPP-conjugated proteins. We screened the library for highly efficient CPPs with no significant cytotoxicity in Escherichia coli and developed a model for predicting the penetration efficiency of a query peptide, enabling the design of new and efficient CPPs. As a proof-of-concept, we used the CPPs for plasmid curing in E. coli and marker gene excision in Methylomonas sp. DH-1. In summary, we demonstrated the utility of CPPs in bacterial engineering. The use of CPPs would facilitate bacterial biotechnology such as genetic engineering, synthetic biology, metabolic engineering, and physiology studies.
在细菌生物技术中，通常不是从质粒生产功能性蛋白质，而是常常需要将功能性蛋白质直接递送到活细胞中进行基因操作或生理修饰。我们构建了一个能够将蛋白质货物递送到细菌中的细胞穿透肽（CPP）文库，并开发了一种用于CPP偶联蛋白的高效递送方法。我们在文库中筛选了对大肠杆菌无明显细胞毒性的高效CPP，并开发了一个预测查询肽穿透效率的模型，从而能够设计新的高效CPP。作为概念验证，我们将CPP用于大肠杆菌中的质粒消除以及甲基单胞菌属DH-1中的标记基因切除。总之，我们证明了CPP在细菌工程中的实用性。CPP的使用将促进细菌生物技术，如基因工程、合成生物学、代谢工程和生理学研究。
