Media Robustness and Scalability of Phosphate Regulated Promoters Useful for Two-Stage Autoinduction in E. coli.

Published

Journal Article

A key challenge in synthetic biology is the successful utilization of characterized parts, such as promoters, in different biological contexts. We report the evaluation of the media robustness of a small library of E. coli PhoB regulated promoters that enable heterologous protein production in two-stage cultures. Expression levels were measured both in a rich Autoinduction Broth as well as a minimal mineral salts media. Expression was both media and promoter dependent. Of the 16 promoters tested, 4 were identified to have tightly controlled expression, which was also robust to media formulation. Improved promoter robustness led to more predictable scale up and consistent expression in instrumented bioreactors. This subset of PhoB activated promoters, useful for two-stage autoinduction, highlights the impact of the environment on the performance of biological parts and the importance of robustness testing in synthetic biology.

Full Text

Duke Authors

Cited Authors

  • Moreb, EA; Ye, Z; Efromson, JP; Hennigan, JN; Menacho-Melgar, R; Lynch, MD

Published Date

  • June 2020

Published In

Volume / Issue

  • 9 / 6

Start / End Page

  • 1483 - 1486

PubMed ID

  • 32353228

Pubmed Central ID

  • 32353228

Electronic International Standard Serial Number (EISSN)

  • 2161-5063

International Standard Serial Number (ISSN)

  • 2161-5063

Digital Object Identifier (DOI)

  • 10.1021/acssynbio.0c00182

Language

  • eng