Plasmid‐encoded protein attenuates Escherichiacoli swimming velocity and cell growth, not reprogrammed regulatory functions

Abstract

In addition to engineering new pathways for synthesis, synthetic biologists rewire cells to carry out “programmable” functions, an example being the creation of wound‐healing probiotics. Engineering regulatory circuits and synthetic machinery, however, can be deleterious to cell function, particularly if the “metabolic burden” is significant. Here, a synthetic regulatory circuit previously constructed to direct Escherichia coli to swim toward hydrogen peroxide, a signal of wound generation, was shown to work even with coexpression of antibiotic resistance genes and genes associated with lactose utilization. We found, however, that cotransformation with a second vector constitutively expressing GFP (as a marker) and additionally conferring resistance to kanamycin and tetracycline resulted in slower velocity (Δ~6 μm/s) and dramatically reduced growth rate (Δ > 50%). The additional vector did not, however, alter the run‐and‐tumble ratio or directional characteristics of H2O2–dependent motility. The main impact of this additional burden was limited to slowing cell velocity and growth, suggesting that reprogrammed cell motility by minimally altering native regulatory circuits can be maintained even when extraneous burden is placed on the host cell. © 2019 American Institute of Chemical Engineers Biotechnol. Prog., 35: e2778, 2019.

Document Details

Document Type
Pub Defense Publication
Publication Date
Feb 15, 2019
Source ID
10.1002/btpr.2778

Entities

People

  • Chelsea Virgile
  • Hsuan‐chen Wu
  • Pricila Hauk
  • William E. Bentley

Organizations

  • Defense Threat Reduction Agency
  • Division of Chemical, Bioengineering, Environmental, and Transport Systems
  • Division of Electrical, Communications & Cyber Systems
  • National Institute of Biomedical Imaging and Bioengineering
  • National Institutes of Health
  • University of Maryland

Tags

Fields of Study

  • Biology

Readers

  • Microbial Pathology
  • Molecular Genetics
  • Oncology (Cancer Research).