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Title: Genome engineering of E. coli for improved styrene production and polymerization

Abstract

Microbial production of exogenous organic compounds is challenging as biosynthetic pathways are often complex and produce metabolites that are toxic to the hosts. Biogenic styrene is an example of this problem, which if addressed could result in a more sustainable supply of this important component of the plastics industry. In this study, we engineered Escherichia coli for the production of styrene. We systematically optimized the production capability by first screening different pathway expression levels in E. coli strains. We then further designed and constructed a transcription regulator library targeting 54 genes with 85,420 mutations, and tested this library for increased styrene resistance and production. A series of tolerant mutants not only exhibited improved styrene tolerance but also produced higher styrene concentrations compared to the parent strain. The best producing mutant, ST05 LexA_E45I, produced a 3.45-fold increase in styrene compared to the parent strain. The produced styrene was extracted via gas stripping into dodecane and used in a direct free radical synthesis of polystyrene.

Authors:
 [1];  [1];  [2];  [3];  [2];  [4];  [2];  [3]
  1. Renewable and Sustainable Energy Inst. (RASEI), Boulder, CO (United States)
  2. Univ. of Colorado, Boulder, CO (United States)
  3. Renewable and Sustainable Energy Inst. (RASEI), Boulder, CO (United States); Univ. of Colorado, Boulder, CO (United States)
  4. National Renewable Energy Laboratory (NREL), Golden, CO (United States); Univ. of Colorado, Boulder, CO (United States); Renewable and Sustainable Energy Inst. (RASEI), Boulder, CO (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
U.S. Department of Defense (DOD), Defense Advanced Research Projects Agency (DARPA); USDOE
OSTI Identifier:
1567025
Report Number(s):
NREL/JA-5400-74993
Journal ID: ISSN 1096-7176
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Accepted Manuscript
Journal Name:
Metabolic Engineering
Additional Journal Information:
Journal Name: Metabolic Engineering; Journal ID: ISSN 1096-7176
Publisher:
Elsevier
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; styrene; transcriptional regulators; genome engineering; polymerization

Citation Formats

Liang, Liya, Liu, Rongming, Foster, Kyle E. O., Choudhury, Alaksh, Cook, Sherri, Cameron, Jeffrey, Srubar III, Wil V., and Gill, Ryan T. Genome engineering of E. coli for improved styrene production and polymerization. United States: N. p., 2019. Web. doi:10.1016/j.ymben.2019.09.007.
Liang, Liya, Liu, Rongming, Foster, Kyle E. O., Choudhury, Alaksh, Cook, Sherri, Cameron, Jeffrey, Srubar III, Wil V., & Gill, Ryan T. Genome engineering of E. coli for improved styrene production and polymerization. United States. doi:10.1016/j.ymben.2019.09.007.
Liang, Liya, Liu, Rongming, Foster, Kyle E. O., Choudhury, Alaksh, Cook, Sherri, Cameron, Jeffrey, Srubar III, Wil V., and Gill, Ryan T. Sun . "Genome engineering of E. coli for improved styrene production and polymerization". United States. doi:10.1016/j.ymben.2019.09.007.
@article{osti_1567025,
title = {Genome engineering of E. coli for improved styrene production and polymerization},
author = {Liang, Liya and Liu, Rongming and Foster, Kyle E. O. and Choudhury, Alaksh and Cook, Sherri and Cameron, Jeffrey and Srubar III, Wil V. and Gill, Ryan T.},
abstractNote = {Microbial production of exogenous organic compounds is challenging as biosynthetic pathways are often complex and produce metabolites that are toxic to the hosts. Biogenic styrene is an example of this problem, which if addressed could result in a more sustainable supply of this important component of the plastics industry. In this study, we engineered Escherichia coli for the production of styrene. We systematically optimized the production capability by first screening different pathway expression levels in E. coli strains. We then further designed and constructed a transcription regulator library targeting 54 genes with 85,420 mutations, and tested this library for increased styrene resistance and production. A series of tolerant mutants not only exhibited improved styrene tolerance but also produced higher styrene concentrations compared to the parent strain. The best producing mutant, ST05 LexA_E45I, produced a 3.45-fold increase in styrene compared to the parent strain. The produced styrene was extracted via gas stripping into dodecane and used in a direct free radical synthesis of polystyrene.},
doi = {10.1016/j.ymben.2019.09.007},
journal = {Metabolic Engineering},
number = ,
volume = ,
place = {United States},
year = {2019},
month = {9}
}

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This content will become publicly available on September 1, 2020
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