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Original Research ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Microbiol. | doi: 10.3389/fmicb.2019.02534

Abiotic factors promote cell penetrating peptide permeability in Enterobacteriaceae models

Daichi Toyohara1, Yasuhito Yokoi1, Go Inoue2, Takahiro Muraoka2 and  Tetsushi Mori1*
  • 1Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Japan
  • 2Department of Organic and Polymer Materials Chemistry, Tokyo University of Agriculture and Technology, Japan

Conventionally, the delivery of biomolecules into bacteria for the generation of characterized or functional mutants have relied greatly on horizontal gene transfer techniques. However, the low compatibility of these techniques with novel or hard-to-transform bacteria currently serves as a challenge to the bioengineering field. Here, we explored the use of cell penetrating peptides (CPPs) as an alternative biomolecule delivery approach by investigating the effects of the abiotic factors during CPP permeation. Using the (KFF)3K-FAM conjugate and Escherichia coli as models, we evaluated four abiotic factors where two of these factors, temperature and solution tonicity promoted (KFF)3K-FAM permeation efficiency. Our data show that optimal (KFF)3K-FAM permeation efficiency was achieved for E. coli at approximately 98.1% under conditions of 37°C (growth optimal temperature) and 50% PBS concentration. Based on these conditions, we subsequently tested the applicability of CPP permeation in various bacterial strains by treating 10 bacterial strains from the Enterobacteriaceae family among which 7 strains have no CPP permeation records with (KFF)3K-FAM. Interestingly, when compared with non-optimized conditions, all 10 strains showed a marked increase in CPP permeation ranging between 20 - 90% efficiency. Although using strains within Enterobacteriaceae that are phylogenetically close, our results hinted on the possibility that with proper optimized of the abiotic factors, CPPs could be compatible with a broad range of bacterial strains. Our efforts suggest that CPP could serve as an effective alternative approach for mutant generation and for biomolecule delivery into novel or hard-to-transform bacteria.

Keywords: Cell-Penetrating Peptides, temperature, solution tonicity, CPP permeation, ABIOTIC factors

Received: 02 May 2019; Accepted: 21 Oct 2019.

Copyright: © 2019 Toyohara, Yokoi, Inoue, Muraoka and Mori. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Prof. Tetsushi Mori, Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, Koganei, Japan, moritets@go.tuat.ac.jp