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Title: Reassessing Escherichia coli as a cell factory for biofuel production

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Journal Article: Publisher's Accepted Manuscript
Journal Name:
Current Opinion in Biotechnology
Additional Journal Information:
Journal Volume: 45; Journal Issue: C; Related Information: CHORUS Timestamp: 2018-01-11 21:37:55; Journal ID: ISSN 0958-1669
Country of Publication:
United Kingdom

Citation Formats

Wang, Chonglong, Pfleger, Brian F., and Kim, Seon-Won. Reassessing Escherichia coli as a cell factory for biofuel production. United Kingdom: N. p., 2017. Web. doi:10.1016/j.copbio.2017.02.010.
Wang, Chonglong, Pfleger, Brian F., & Kim, Seon-Won. Reassessing Escherichia coli as a cell factory for biofuel production. United Kingdom. doi:10.1016/j.copbio.2017.02.010.
Wang, Chonglong, Pfleger, Brian F., and Kim, Seon-Won. 2017. "Reassessing Escherichia coli as a cell factory for biofuel production". United Kingdom. doi:10.1016/j.copbio.2017.02.010.
title = {Reassessing Escherichia coli as a cell factory for biofuel production},
author = {Wang, Chonglong and Pfleger, Brian F. and Kim, Seon-Won},
abstractNote = {},
doi = {10.1016/j.copbio.2017.02.010},
journal = {Current Opinion in Biotechnology},
number = C,
volume = 45,
place = {United Kingdom},
year = 2017,
month = 6

Journal Article:
Free Publicly Available Full Text
This content will become publicly available on March 11, 2018
Publisher's Accepted Manuscript

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  • Biological production of chemicals and fuels using microbial transformation of sustainable carbon sources, such as pretreated and saccharified plant biomass, is a multi-step process. Typically, each segment of the workflow is optimized separately, often generating conditions that may not be suitable for integration or consolidation with the upstream or downstream steps. While significant effort has gone into developing solutions to incompatibilities at discrete steps, very few studies report the consolidation of the multi-step workflow into a single pot reactor system. Here we demonstrate a one-pot biofuel production process that uses the ionic liquid 1-ethyl-3-methylimidazolium acetate (C 2C 1Im][OAc] ) formore » pretreatment of switchgrass biomass. [C 2C 1Im][OAc] is highly effective in deconstructing lignocellulose, but nonetheless leaves behind residual reagents that are toxic to standard saccharification enzymes and the microbial production host. We report the discovery of an [C 2C 1Im]-tolerant E. coli strain, where [C 2C 1Im] tolerance is bestowed by a P7Q mutation in the transcriptional regulator encoded by rcdA. We establish that the causal impact of this mutation is the derepression of a hitherto uncharacterized major facilitator family transporter, YbjJ. To develop the strain for a one-pot process we engineered this [C 2C 1Im]-tolerant strain to express a recently reported d-limonene production pathway. We also screened previously reported [C 2C 1Im]-tolerant cellulases to select one that would function with the range of E. coli cultivation conditions and expressed it in the [C 2C 1 Im]-tolerant E. coli strain so as to secrete this [C 2C 1Im]-tolerant cellulase. The final strain digests pretreated biomass, and uses the liberated sugars to produce the bio-jet fuel candidate precursor d-limonene in a one-pot process.« less
  • Isolated Escherichia coli K-12 cell envelopes or Bacillus subtilis 168 cell walls were reacted with smectite or kaolinite clay in distilled deionized water (pH 6.0); unbound envelopes or walls were separated by sucrose density gradient centrifugation, and the extent of adsorption was calculated. The metal-binding capacity of isolated envelopes, walls, clays, and envelope-clay or wall-clay mixtures was determined by atomic absorption spectroscopy after exposure to aqueous 5.0 mM Ag{sup +}, Cu{sup 2+}, Cd{sup 2+}, Ni{sup 2+}, Pb{sup 2+}, Zn{sup 2+}, and Cr{sup 3+} nitrate salt solutions at pHs determined by the buffering capacity of wall, envelope, clay, or composite system.more » The order of metal uptake was walls > envelopes > smectite clay > kaolinite clay for the individual components, and walls plus smectite > walls plus kaolinite > envelopes plus smectite > envelopes plus kaolinite for the mixtures. On a dry-weight basis, the envelope-clay and wall-clay mixtures bound 20 to 90% less metal than equal amounts of the individual components did. This reduction in metal-binding capacity indicates that the adsorption of the wall or envelope to clay has masked or neutralized chemically reactive adsorption sites normally available to metal ions.« less