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Title: Substantial improvements in methyl ketone production in E. coli and insights on the pathway from in vitro studies

Authors:
; ; ; ; ;
Publication Date:
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1222408
Grant/Contract Number:
AC02-05CH11231
Resource Type:
Journal Article: Publisher's Accepted Manuscript
Journal Name:
Metabolic Engineering
Additional Journal Information:
Journal Volume: 26; Journal Issue: C; Related Information: CHORUS Timestamp: 2016-03-26 19:50:23; Journal ID: ISSN 1096-7176
Publisher:
Elsevier
Country of Publication:
Belgium
Language:
English

Citation Formats

Goh, Ee-Been, Baidoo, Edward E. K., Burd, Helcio, Lee, Taek Soon, Keasling, Jay D., and Beller, Harry R.. Substantial improvements in methyl ketone production in E. coli and insights on the pathway from in vitro studies. Belgium: N. p., 2014. Web. doi:10.1016/j.ymben.2014.09.003.
Goh, Ee-Been, Baidoo, Edward E. K., Burd, Helcio, Lee, Taek Soon, Keasling, Jay D., & Beller, Harry R.. Substantial improvements in methyl ketone production in E. coli and insights on the pathway from in vitro studies. Belgium. doi:10.1016/j.ymben.2014.09.003.
Goh, Ee-Been, Baidoo, Edward E. K., Burd, Helcio, Lee, Taek Soon, Keasling, Jay D., and Beller, Harry R.. Sat . "Substantial improvements in methyl ketone production in E. coli and insights on the pathway from in vitro studies". Belgium. doi:10.1016/j.ymben.2014.09.003.
@article{osti_1222408,
title = {Substantial improvements in methyl ketone production in E. coli and insights on the pathway from in vitro studies},
author = {Goh, Ee-Been and Baidoo, Edward E. K. and Burd, Helcio and Lee, Taek Soon and Keasling, Jay D. and Beller, Harry R.},
abstractNote = {},
doi = {10.1016/j.ymben.2014.09.003},
journal = {Metabolic Engineering},
number = C,
volume = 26,
place = {Belgium},
year = {Sat Nov 01 00:00:00 EDT 2014},
month = {Sat Nov 01 00:00:00 EDT 2014}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record at 10.1016/j.ymben.2014.09.003

Citation Metrics:
Cited by: 16works
Citation information provided by
Web of Science

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  • Previously, we developed an E. coli strain that overproduces medium-chain methyl ketones for potential use as diesel fuel blending agents or as flavors and fragrances. To date, the strain's performance has been optimized during growth with glucose. However, lignocellulosic biomass hydrolysates also contain a substantial portion of hemicellulose-derived xylose, which is typically the second most abundant sugar after glucose. Commercialization of the methyl ketone-producing technology would benefit from the increased efficiency resulting from simultaneous, rather than the native sequential (diauxic), utilization of glucose and xylose. In this study, genetic manipulations were performed to alleviate carbon catabolite repression in our mostmore » efficient methyl ket one-producing strain. A strain engineered for constitutive expression of xylF and xylA (involved in xylose transport and metabolism) showed synchronized glucose and xylose consumption rates. However, this newly acquired capability came at the expense of methyl ketone titer, which decreased fivefold. Further efforts were made to improve methyl ketone production in this strain, and we found that two strategies were effective at enhancing methyl ketone titer: (1) chromosomal deletion of pgi (glucose-6-phosphate isomerase) to increase intracellular NADPH supply and (2) downregulation of CRP (cAMP receptor protein) expression by replacement of the native RBS with an RBS chosen based upon mutant library screening results. Combining these strategies resulted in the most favorable overall phenotypes for simultaneous glucose-xylose consumption without compromising methyl ketone titer at both 1 and 2% total sugar concentrations in shake flasks. This work demonstrated a strategy for engineering simultaneous utilization of C 6 and C 5 sugars in E. coli without sacrificing production of fatty acid-derived compounds.« less
  • A method is proposed for the synthesis of methyl vinyl ketone by hydration trimerization of acetylene followed by hydrolysis of the 2-vinloxy-1,3-butadiene formed under the action of a potassium hydroxide-potassium phosphate-DMSO catalyst system. The authors contend that this system eliminates the normal drawbacks associated with synthesis of the ketone, including low product yield, multistage operations, the presence of impurities that are difficult to remove, and high cost. The proposed method results in a highly pure product obtained in high yield.
  • The conditions of stabilization of the perruthenate ion in solutions of sodium hydroxide in the presence of sodium hypochlorite were found. It was shown that at various alkali concentrations there are the corresponding concentrations of hypochlorite, below which only the perruthenate ion exists, and ruthenium tetraoxide is not formed. With the aid of ruthenium-106 the extraction of ruthenium (VII) with TBP and methyl ethyl ketone (MEK) was studied. In extraction with MEK a comparatively rapid reduction of the perruthenate ion and the formation of a precipitate RuO/sub 2/ . nH/sub 2/O are observed. In the case of extraction with TBP,more » even for 24 h, there is no reduction of ruthenium (VII), either in the aqueous or in the organic phase. By determining the distribution coefficients of sodium (/sup 22/Na was used) in the absence and in the presence of various amounts of ruthenium (VII) and by measuring the absorption spectra of the initial solutions and extracts, it was shown that ruthenium is extracted in the range of its concentrations 10/sup -9/-5 . 10/sup -4/ M TBP in the form of hydrate-solvates of the acid HRuO/sub 4/ and the salt NaRuO/sub 4/.« less