Identification of efficient prokaryotic cell-penetrating peptides with applications in bacterial biotechnology

Commun Biol. 2021 Feb 15;4(1):205. doi: 10.1038/s42003-021-01726-w.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotechnology*
  • CHO Cells
  • Cell-Penetrating Peptides / genetics
  • Cell-Penetrating Peptides / metabolism*
  • Cricetulus
  • Electroporation
  • Escherichia coli / genetics
  • Escherichia coli / metabolism*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Gene Expression Regulation, Bacterial
  • Genetic Engineering
  • HEK293 Cells
  • Humans
  • Industrial Microbiology*
  • Methylomonas / genetics
  • Methylomonas / metabolism*
  • Peptide Library
  • Plasmids / genetics
  • Plasmids / metabolism
  • Proof of Concept Study
  • Protein Transport

Substances

  • Cell-Penetrating Peptides
  • Escherichia coli Proteins
  • Peptide Library