A bimodular PKS platform that expands the biological design space

Zargar, Amin; Valencia, Luis; Wang, Jessica; Lal, Ravi; Chang, Samantha; Werts, Miranda; Wong, Andrew R.; Hernández, Amanda C.; Benites, Veronica; Baidoo, Edward E.K.; Katz, Leonard; Keasling, Jay D.

doi:10.1016/j.ymben.2020.07.001

Title: A bimodular PKS platform that expands the biological design space

Journal Article · Thu Aug 06 00:00:00 EDT 2020 · Metabolic Engineering

DOI:https://doi.org/10.1016/j.ymben.2020.07.001· OSTI ID:1765567

Zargar, Amin ^[1]; Valencia, Luis ^[2];

^[3];

^[3]; Chang, Samantha ^[3]; Werts, Miranda ^[3]; Wong, Andrew R. ^[3];

^[3]; Benites, Veronica ^[4]; Baidoo, Edward E.K. ^[4]; Katz, Leonard ^[5]; Keasling, Jay D. ^[6]

Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California-Berkeley, Emeryville, CA (United States). QB3 Inst.
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States)
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Dept. of Energy Agile BioFoundry, Emeryville, CA (United States)
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Univ. of California-Berkeley, Emeryville, CA (United States). QB3 Inst.
Joint BioEnergy Institute (JBEI), Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California-Berkeley, Emeryville, CA (United States). QB3 Inst.; Univ. of California, Berkeley, CA (United States); Technical Univ., Horsholm (Denmark). Novo Nordisk Foundation Center; Shenzhen Inst. for Advanced Technologies (China). Synthetic Biochemistry Center

Traditionally engineered to produce novel bioactive molecules, Type I modular polyketide synthases (PKSs) could be engineered as a new biosynthetic platform for the production of de novo fuels, commodity chemicals, and specialty chemicals. Previously, our investigations manipulated the first module of the lipomycin PKS to produce short chain ketones, 3-hydroxy acids, and saturated, branched carboxylic acids. Building upon this work, we have expanded to multi-modular systems by engineering the first two modules of lipomycin to generate unnatural polyketides as potential biofuels and specialty chemicals in Streptomyces albus. First, we produce 20.6 mg/L of the ethyl ketone, 4,6 dimethylheptanone through a reductive loop exchange in LipPKS1 and a ketoreductase knockouts in LipPKS2. Furthermore, we then show that an AT swap in LipPKS1 and a reductive loop exchange in LipPKS2 can produce the potential fragrance 3-isopropyl-6-methyltetrahydropyranone. Highlighting the challenge of maintaining product fidelity, in both bimodular systems we observed side products from premature hydrolysis in the engineered first module and stalled dehydration in reductive loop exchanges. Collectively, our work expands the biological design space and moves the field closer to the production of "designer" biomolecules.

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

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH)

Grant/Contract Number:: AC02-05CH11231; F32GM125179

OSTI ID:: 1765567

Alternate ID(s):: OSTI ID: 1650187

Journal Information:: Metabolic Engineering, Vol. 61; ISSN 1096-7176

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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