Adaptive laboratory evolution and transcriptomics-guided engineering of Escherichia coli for increased isobutanol tolerance

Young Seo Jang, Jungwoo Yang, Jae Kyun Kim, Tae In Kim, Yong Cheol Park, In Jung Kim, Kyoung Heon Kim

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

As a renewable energy from biomass, isobutanol is considered as a promising alternative to fossil fuels. To biotechnologically produce isobutanol, strain development using industrial microbial hosts, such as Escherichia coli, has been conducted by introducing a heterologous isobutanol synthetic pathway. However, the toxicity of produced isobutanol inhibits cell growth, thereby restricting improvements in isobutanol titer, yield, and productivity. Therefore, the development of robust microbial strains tolerant to isobutanol is required. In this study, isobutanol-tolerant mutants were isolated from two E. coli parental strains, E. coli BL21(DE3) and MG1655(DE3), through adaptive laboratory evolution (ALE) under high isobutanol concentrations. Subsequently, 16 putative genes responsible for isobutanol tolerance were identified by transcriptomic analysis. When overexpressed in E. coli, four genes (fadB, dppC, acs, and csiD) conferred isobutanol tolerance. A fermentation study with a reverse engineered isobutanol-producing E. coli JK209 strain showed that fadB or dppC overexpression improved isobutanol titers by 1.5 times, compared to the control strain. Through coupling adaptive evolution with transcriptomic analysis, new genetic targets utilizable were identified as the basis for the development of an isobutanol-tolerant strain. Thus, these new findings will be helpful not only for a fundamental understanding of microbial isobutanol tolerance but also for facilitating industrially feasible isobutanol production.

Original languageEnglish
Article number2300270
JournalBiotechnology Journal
Volume19
Issue number1
DOIs
Publication statusPublished - 2024 Jan

Bibliographical note

Publisher Copyright:
© 2023 Wiley-VCH GmbH.

Keywords

  • adaptive laboratory evolution
  • evolutionary engineering
  • isobutanol
  • isobutanol tolerance
  • reverse engineering
  • transcriptomics

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Molecular Medicine

Fingerprint

Dive into the research topics of 'Adaptive laboratory evolution and transcriptomics-guided engineering of Escherichia coli for increased isobutanol tolerance'. Together they form a unique fingerprint.

Cite this