Engineering Saccharomyces cerevisiae for isoprenol production

Area of Science:

• Biotechnology
• Metabolic Engineering
• Synthetic Biology

Background:

• Isoprenol is a key biofuel and chemical precursor.
• Escherichia coli has been engineered for isoprenol production.
• Saccharomyces cerevisiae has potential for isoprenoid production but lacks established isoprenol biosynthesis.

Purpose of the Study:

• To engineer Saccharomyces cerevisiae for efficient isoprenol biosynthesis.
• To improve isoprenol production titers in yeast.
• To establish yeast as an industrial platform for isoprenol.

Main Methods:

• Heterologous mevalonate pathway construction.
• Implementation of an isopentenyl diphosphate (IPP)-bypass pathway.
• Deletion of a promiscuous endogenous kinase.
• Metabolomics analysis to identify pathway bottlenecks.
• Overexpression of a promiscuous alkaline phosphatase.

Main Results:

• Initial isoprenol production of 36.02 mg/L using the mevalonate pathway.
• The IPP-bypass pathway doubled the isoprenol titer.
• Strain engineering, including kinase deletion, increased titer to 130.52 mg/L.
• Metabolomics-guided overexpression of alkaline phosphatase resulted in a final titer of 383.1 mg/L.
• This represents the highest isoprenol titer reported in S. cerevisiae to date.

Conclusions:

• Saccharomyces cerevisiae can be effectively engineered for isoprenol production.
• Pathway engineering strategies, including bypass pathways and bottleneck relief, significantly enhance titers.
• This work provides a foundation for yeast-based industrial isoprenol manufacturing.