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Title: Engineering Corynebacterium glutamicum to produce the biogasoline isopentenol from plant biomass hydrolysates

Abstract

Background:Many microbes used for the rapid discovery and development of metabolic pathways have sensitivities to final products and process reagents. Isopentenol (3-methyl-3-buten-1-ol), a biogasoline candidate, has an established heterologous gene pathway but is toxic to several microbial hosts. Reagents used in the pretreatment of plant biomass, such as ionic liquids, also inhibit growth of many host strains. We explored the use of Corynebacterium glutamicum as an alternative host to address these constraints. Results:We found C. glutamicum ATCC 13032 to be tolerant to both the final product, isopentenol, as well to three classes of ionic liquids. A heterologous mevalonate-based isopentenol pathway was engineered in C. glutamicum. Targeted proteomics for the heterologous pathway proteins indicated that the 3-hydroxy-3-methylglutaryl-coenzyme A reductase protein, HmgR, is a potential rate-limiting enzyme in this synthetic pathway. Isopentenol titers were improved from undetectable to 1.25 g/L by combining three approaches: media optimization; substitution of an NADH-dependent HmgR homolog from Silicibacter pomeroyi; and development of a C. glutamicum ΔpoxB ΔldhA host chassis. Conclusions:We describe the successful expression of a heterologous mevalonate-based pathway in the Gram-positive industrial microorganism, C. glutamicum, for the production of the biogasoline candidate, isopentenol. We identified critical genetic factors to harness the isopentenol pathway in C. glutamicum.more » Further media and cultivation optimization enabled isopentenol production from sorghum biomass hydrolysates.« less

Authors:
; ; ; ; ; ; ; ORCiD logo; ; ORCiD logo
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1618759
Alternate Identifier(s):
OSTI ID: 1545316
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Published Article
Journal Name:
Biotechnology for Biofuels
Additional Journal Information:
Journal Name: Biotechnology for Biofuels Journal Volume: 12 Journal Issue: 1; Journal ID: ISSN 1754-6834
Publisher:
Springer Science + Business Media
Country of Publication:
Netherlands
Language:
English
Subject:
09 BIOMASS FUELS

Citation Formats

Sasaki, Yusuke, Eng, Thomas, Herbert, Robin A., Trinh, Jessica, Chen, Yan, Rodriguez, Alberto, Gladden, John, Simmons, Blake A., Petzold, Christopher J., and Mukhopadhyay, Aindrila. Engineering Corynebacterium glutamicum to produce the biogasoline isopentenol from plant biomass hydrolysates. Netherlands: N. p., 2019. Web. doi:10.1186/s13068-019-1381-3.
Sasaki, Yusuke, Eng, Thomas, Herbert, Robin A., Trinh, Jessica, Chen, Yan, Rodriguez, Alberto, Gladden, John, Simmons, Blake A., Petzold, Christopher J., & Mukhopadhyay, Aindrila. Engineering Corynebacterium glutamicum to produce the biogasoline isopentenol from plant biomass hydrolysates. Netherlands. doi:10.1186/s13068-019-1381-3.
Sasaki, Yusuke, Eng, Thomas, Herbert, Robin A., Trinh, Jessica, Chen, Yan, Rodriguez, Alberto, Gladden, John, Simmons, Blake A., Petzold, Christopher J., and Mukhopadhyay, Aindrila. Wed . "Engineering Corynebacterium glutamicum to produce the biogasoline isopentenol from plant biomass hydrolysates". Netherlands. doi:10.1186/s13068-019-1381-3.
@article{osti_1618759,
title = {Engineering Corynebacterium glutamicum to produce the biogasoline isopentenol from plant biomass hydrolysates},
author = {Sasaki, Yusuke and Eng, Thomas and Herbert, Robin A. and Trinh, Jessica and Chen, Yan and Rodriguez, Alberto and Gladden, John and Simmons, Blake A. and Petzold, Christopher J. and Mukhopadhyay, Aindrila},
abstractNote = {Background:Many microbes used for the rapid discovery and development of metabolic pathways have sensitivities to final products and process reagents. Isopentenol (3-methyl-3-buten-1-ol), a biogasoline candidate, has an established heterologous gene pathway but is toxic to several microbial hosts. Reagents used in the pretreatment of plant biomass, such as ionic liquids, also inhibit growth of many host strains. We explored the use of Corynebacterium glutamicum as an alternative host to address these constraints. Results:We found C. glutamicum ATCC 13032 to be tolerant to both the final product, isopentenol, as well to three classes of ionic liquids. A heterologous mevalonate-based isopentenol pathway was engineered in C. glutamicum. Targeted proteomics for the heterologous pathway proteins indicated that the 3-hydroxy-3-methylglutaryl-coenzyme A reductase protein, HmgR, is a potential rate-limiting enzyme in this synthetic pathway. Isopentenol titers were improved from undetectable to 1.25 g/L by combining three approaches: media optimization; substitution of an NADH-dependent HmgR homolog from Silicibacter pomeroyi; and development of a C. glutamicum ΔpoxB ΔldhA host chassis. Conclusions:We describe the successful expression of a heterologous mevalonate-based pathway in the Gram-positive industrial microorganism, C. glutamicum, for the production of the biogasoline candidate, isopentenol. We identified critical genetic factors to harness the isopentenol pathway in C. glutamicum. Further media and cultivation optimization enabled isopentenol production from sorghum biomass hydrolysates.},
doi = {10.1186/s13068-019-1381-3},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 12,
place = {Netherlands},
year = {2019},
month = {2}
}

Journal Article:
Free Publicly Available Full Text
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DOI: 10.1186/s13068-019-1381-3

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Cited by: 6 works
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    Works referencing / citing this record:

    Growth-uncoupled isoprenoid synthesis in Rhodobacter sphaeroides
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