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

Sasaki, Yusuke; Eng, Thomas; Herbert, Robin A.; Trinh, Jessica; Chen, Yan; Rodriguez, Alberto; Gladden, John; Simmons, Blake A.; Petzold, Christopher J.; Mukhopadhyay, Aindrila

doi:10.1186/s13068-019-1381-3

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

Journal Article · Wed Feb 27 00:00:00 EST 2019 · Biotechnology for Biofuels

DOI:https://doi.org/10.1186/s13068-019-1381-3· OSTI ID:1618759

Sasaki, Yusuke; Eng, Thomas; Herbert, Robin A.; Trinh, Jessica; Chen, Yan; Rodriguez, Alberto; Gladden, John; ; Petzold, Christopher J.;

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 ∆poxB ∆ldhA 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.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE; USDOE Office of Science (SC)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1618759

Alternate ID(s):: OSTI ID: 1545316

Journal Information:: Biotechnology for Biofuels, Journal Name: Biotechnology for Biofuels Journal Issue: 1 Vol. 12; ISSN 1754-6834

Publisher:: Springer Science + Business MediaCopyright Statement

Country of Publication:: Netherlands

Language:: English

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