Engineering Saccharomyces cerevisiae for isoprenol production

Jinho Kim; Edward E K Baidoo; Bashar Amer; Aindrila Mukhopadhyay; Paul D Adams; Blake A Simmons; Taek Soon Lee

doi:10.1016/j.ymben.2021.02.002

Engineering Saccharomyces cerevisiae for isoprenol production

Metab Eng. 2021 Mar:64:154-166. doi: 10.1016/j.ymben.2021.02.002. Epub 2021 Feb 10.

Authors

Jinho Kim¹, Edward E K Baidoo¹, Bashar Amer¹, Aindrila Mukhopadhyay², Paul D Adams³, Blake A Simmons¹, Taek Soon Lee⁴

Affiliations

¹ Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
² Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA; Environmental Genomics and Systems Biology, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA.
³ Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Molecular Biophysics and Integrated Bioimaging, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA; Department of Bioengineering, University of California, Berkeley, CA 94720, USA.
⁴ Joint BioEnergy Institute, 5885 Hollis Street, Emeryville, CA 94608, USA; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA. Electronic address: tslee@lbl.gov.

PMID: 33581331
DOI: 10.1016/j.ymben.2021.02.002

Abstract

Isoprenol (3-methyl-3-butene-1-ol) is a valuable drop-in biofuel and an important precursor of several commodity chemicals. Synthetic microbial systems using the heterologous mevalonate pathway have recently been developed for the production of isoprenol in Escherichia coli, and a significant yield and titer improvement has been achieved through a decade of research. Saccharomyces cerevisiae has been widely used in the biotechnology industry for isoprenoid production, but there has been no good example of isoprenol production reported in this host. In this study, we engineered the budding yeast S. cerevisiae for improved biosynthesis of isoprenol. The strain engineered with the mevalonate pathway achieved isoprenol production at the titer of 36.02 ± 0.92 mg/L in the flask. The IPP (isopentenyl diphosphate)-bypass pathway, which has shown more efficient isoprenol production by avoiding the accumulation of the toxic intermediate in E. coli, was also constructed in S. cerevisiae and improved the isoprenol titer by 2-fold. We further engineered the strains by deleting a promiscuous endogenous kinase that could divert the pathway flux away from the isoprenol production and improved the titer to 130.52 ± 8.01 mg/L. Finally, we identified a pathway bottleneck using metabolomics analysis and overexpressed a promiscuous alkaline phosphatase to relieve this bottleneck. The combined efforts resulted in the titer improvement to 383.1 ± 31.62 mg/L in the flask. This is the highest isoprenol titer up to date in S. cerevisiae and this work provides the key strategies to engineer yeast as an industrial platform for isoprenol production.

Keywords: Biofuel; IPP-Bypass pathway; Isoprenol; Metabolic engineering; Mevalonate pathway; Saccharomyces cerevisiae.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Escherichia coli / genetics
Escherichia coli Proteins* / genetics
Metabolic Engineering
Mevalonic Acid
Saccharomyces cerevisiae* / genetics

Substances

Escherichia coli Proteins
Mevalonic Acid