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Title: Short-chain ketone production by engineered polyketide synthases in Streptomyces albus

Abstract

Microbial production of fuels and commodity chemicals has been performed primarily using natural or slightly modified enzymes, which inherently limits the types of molecules that can be produced. Type I modular polyketide synthases (PKSs) are multi-domain enzymes that can produce unique and diverse molecular structures by combining particular types of catalytic domains in a specific order. This catalytic mechanism offers a wealth of engineering opportunities. Here we report engineered microbes that produce various short-chain (C5-C7) ketones using hybrid PKSs. Introduction of the genes into the chromosome of Streptomyces albus enables it to produce >1 g · l-1 of C6 and C7 ethyl ketones and several hundred mg · l-1 of C5 and C6 methyl ketones from plant biomass hydrolysates. Engine tests indicate these short-chain ketones can be added to gasoline as oxygenates to increase the octane of gasoline. Together, it demonstrates the efficient and renewable microbial production of biogasolines by hybrid enzymes.

Authors:
ORCiD logo [1];  [2];  [3];  [4];  [2];  [5];  [5];  [2];  [2];  [2];  [6];  [7];  [6]; ORCiD logo [5];  [8]; ORCiD logo [9]
  1. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biogical Systems and Engineering Division; Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of Tokyo (Japan). Biotechnology Research Center
  2. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biogical Systems and Engineering Division, and Advanced Biofuels & Bioproducts Process Development Unit
  3. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biogical Systems and Engineering Division
  4. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); National Inst. of Advanced Industrial Science and Technology, Higashi-hiroshima, Hiroshima (Japan). Research Inst. for Sustainable Chemistry, Inst. for Synthetic Biology
  5. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biogical Systems and Engineering Division; Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States)
  6. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)
  7. Sandia National Lab. (SNL-CA), Livermore, CA (United States)
  8. Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States). QB3 Inst.
  9. Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biogical Systems and Engineering Division; Joint BioEnergy Inst. (JBEI), Emeryville, CA (United States); Univ. of California, Berkeley, CA (United States). QB3 Inst., Dept. of Bioengineering, Dept. of Chemical and Biomolecular Engineering; Technical Univ. of Denmark, Lyngby (Denmark). Novo Nordisk Foundation Center for Biosustainability; Shenzhen Inst. for Advanced Technologies, Shenzhen, Guangdong (China). Center for Synthetic Biochemistry
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER); National Science Foundation (NSF)
OSTI Identifier:
1493265
Grant/Contract Number:  
AC02-05CH11231
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY

Citation Formats

Yuzawa, Satoshi, Mirsiaghi, Mona, Jocic, Renee, Fujii, Tatsuya, Masson, Fabrice, Benites, Veronica T., Baidoo, Edward E. K., Sundstrom, Eric, Tanjore, Deepti, Pray, Todd R., George, Anthe, Davis, Ryan W., Gladden, John M., Simmons, Blake A., Katz, Leonard, and Keasling, Jay D. Short-chain ketone production by engineered polyketide synthases in Streptomyces albus. United States: N. p., 2018. Web. doi:10.1038/s41467-018-07040-0.
Yuzawa, Satoshi, Mirsiaghi, Mona, Jocic, Renee, Fujii, Tatsuya, Masson, Fabrice, Benites, Veronica T., Baidoo, Edward E. K., Sundstrom, Eric, Tanjore, Deepti, Pray, Todd R., George, Anthe, Davis, Ryan W., Gladden, John M., Simmons, Blake A., Katz, Leonard, & Keasling, Jay D. Short-chain ketone production by engineered polyketide synthases in Streptomyces albus. United States. doi:10.1038/s41467-018-07040-0.
Yuzawa, Satoshi, Mirsiaghi, Mona, Jocic, Renee, Fujii, Tatsuya, Masson, Fabrice, Benites, Veronica T., Baidoo, Edward E. K., Sundstrom, Eric, Tanjore, Deepti, Pray, Todd R., George, Anthe, Davis, Ryan W., Gladden, John M., Simmons, Blake A., Katz, Leonard, and Keasling, Jay D. Thu . "Short-chain ketone production by engineered polyketide synthases in Streptomyces albus". United States. doi:10.1038/s41467-018-07040-0. https://www.osti.gov/servlets/purl/1493265.
@article{osti_1493265,
title = {Short-chain ketone production by engineered polyketide synthases in Streptomyces albus},
author = {Yuzawa, Satoshi and Mirsiaghi, Mona and Jocic, Renee and Fujii, Tatsuya and Masson, Fabrice and Benites, Veronica T. and Baidoo, Edward E. K. and Sundstrom, Eric and Tanjore, Deepti and Pray, Todd R. and George, Anthe and Davis, Ryan W. and Gladden, John M. and Simmons, Blake A. and Katz, Leonard and Keasling, Jay D.},
abstractNote = {Microbial production of fuels and commodity chemicals has been performed primarily using natural or slightly modified enzymes, which inherently limits the types of molecules that can be produced. Type I modular polyketide synthases (PKSs) are multi-domain enzymes that can produce unique and diverse molecular structures by combining particular types of catalytic domains in a specific order. This catalytic mechanism offers a wealth of engineering opportunities. Here we report engineered microbes that produce various short-chain (C5-C7) ketones using hybrid PKSs. Introduction of the genes into the chromosome of Streptomyces albus enables it to produce >1 g · l-1 of C6 and C7 ethyl ketones and several hundred mg · l-1 of C5 and C6 methyl ketones from plant biomass hydrolysates. Engine tests indicate these short-chain ketones can be added to gasoline as oxygenates to increase the octane of gasoline. Together, it demonstrates the efficient and renewable microbial production of biogasolines by hybrid enzymes.},
doi = {10.1038/s41467-018-07040-0},
journal = {Nature Communications},
number = 1,
volume = 9,
place = {United States},
year = {2018},
month = {11}
}

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Figures / Tables:

Figure 1 Figure 1: Hybrid polyketide synthases engineered to produce short-chain ketones. Short-chain ketone-producing polyketide synthases (PKSs) were created by engineering of LipPks1, a type I modular PKS. Catalytic domains of LipPks1 are shown in green. The non-catalytic domains are shown in gray. Linkers are shown as black line. The TE domainmore » (blue) is derived from the erythromycin PKS. The AT domain shown in magenta is derived from the borrelidin PKS module 1, which is specific for malonyl-CoA. Enzymatic products that can be produced by each PKS were shown in the same box. AT acyltransferase, ACP acyl carrier protein, CoA coenzyme A, KAL KS to AT linker, KR ketoreductase, KS ketosynthetas, PAL Post AT linker, TE thioesterase« less

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