Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei

Tomko, Timothy A.; Dunlop, Mary J.

doi:10.1186/s13068-015-0347-3

Title: Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei

Journal Article · 07 October 2015 · Biotechnology for Biofuels

DOI: https://doi.org/10.1186/s13068-015-0347-3 · OSTI ID:1618620

Tomko, Timothy A. ^[1]; Dunlop, Mary J. ^[1]

Univ. of Vermont, Burlington, VT (United States). School of Engineering

Background: Recent metabolic engineering efforts have generated microorganisms that can produce biofuels, including bio-jet fuels, however these fuels are often toxic to cells, limiting production yields. There are natural examples of microorganisms that have evolved mechanisms for tolerating hydrocarbon-rich environments, such as those that thrive near natural oil seeps and in oil-polluted waters. Results: Using genomic DNA from the hydrocarbon-degrading microbe Marinobacter aquaeolei, we constructed a transgenic library that we expressed in Escherichia coli. We exposed cells to inhibitory levels of pinene, a monoterpene that can serve as a jet fuel precursor with chemical properties similar to existing tactical fuels. Using a sequential strategy with a fosmid library followed by a plasmid library, we were able to isolate a region of DNA from the M. aquaeolei genome that conferred pinene tolerance when expressed in E. coli. We determined that a single gene, yceI, was responsible for the tolerance improvements. Overexpression of this gene placed no additional burden on the host. We also tested tolerance to other monoterpenes and showed that yceI selectively improves tolerance. Conclusions: The genomes of hydrocarbon-tolerant microbes represent a rich resource for tolerance engineering. Using a transgenic library, we were able to identify a single gene that improves E. coli’s tolerance to the bio-jet fuel precursor pinene.

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Research Organization:: Univ. of Vermont, Burlington, VT (United States)

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

Grant/Contract Number:: FG02-00ER41132

OSTI ID:: 1618620

Alternate ID(s):: OSTI ID: 1626967

Journal Information:: Biotechnology for Biofuels, Vol. 8, Issue 1; ISSN 1754-6834

Publisher:: BioMed CentralCopyright Statement

Country of Publication:: United States

Language:: English

Citation Metrics:

Cited by: 19 works

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Related Subjects

09 BIOMASS FUELS
Biotechnology & Applied Microbiology
Energy & Fuels
Biofuel tolerance
Marinobacter aquaeolei
YceI
Pinene
Monoterpene
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Transgenic

MOESM2 of Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791_d3 The current record is supplemented by this text	text	January 2015
MOESM1 of Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791_d2 The current record is supplemented by this text	text	January 2015
MOESM3 of Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791_d1 The current record is supplemented by this text	text	January 2015
Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791 The current record is supplemented by this collection	collection	January 2015
MOESM3 of Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791_d1.v1 The current record is supplemented by this text	text	January 2015
Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791.v1 The current record is supplemented by this collection	collection	January 2015
MOESM1 of Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791_d2.v1 The current record is supplemented by this text	text	January 2015
MOESM2 of Engineering improved bio-jet fuel tolerance in Escherichia coli using a transgenic library from the hydrocarbon-degrader Marinobacter aquaeolei Tomko, Timothy; Dunlop, Mary https://doi.org/10.6084/m9.figshare.c.3627791_d3.v1 The current record is supplemented by this text	text	January 2015