Production of 10-methyl branched fatty acids in yeast
Journal article, 2021

Background: Despite the environmental value of biobased lubricants, they account for less than 2% of global lubricant use due to poor thermo-oxidative stability arising from the presence of unsaturated double bonds. Methyl branched fatty acids (BFAs), particularly those with branching near the acyl-chain mid-point, are a high-performance alternative to existing vegetable oils because of their low melting temperature and full saturation.
Results: We cloned and characterized two pathways to produce 10-methyl BFAs isolated from actinomycetes and γ-proteobacteria. In the two-step bfa pathway of actinomycetes, BfaB methylates Δ9 unsaturated fatty acids to form 10-methylene BFAs, and subsequently, BfaA reduces the double bond to produce a fully saturated 10-methyl branched fatty acid. A BfaA-B fusion enzyme increased the conversion efficiency of 10-methyl BFAs. The ten-methyl palmitate production (tmp) pathway of γ-proteobacteria produces a 10-methylene intermediate, but the TmpA putative reductase was not active in E. coli or yeast. Comparison of BfaB and TmpB activities revealed a range of substrate specificities from C14-C20 fatty acids unsaturated at the Δ9, Δ10 or Δ11 position. We demonstrated efficient production of 10-methylene and 10-methyl BFAs in S. cerevisiae by secretion of free fatty acids and in Y. lipolytica as triacylglycerides, which accumulated to levels more than 35% of total cellular fatty acids.
Conclusions: We report here the characterization of a set of enzymes that can produce position-specific methylene and methyl branched fatty acids. Yeast expression of bfa enzymes can provide a platform for the large-scale production of branched fatty acids suitable for industrial and consumer applications.

Biobased lubricant

10-Methylstearic acid

Yarrowia lipolytica

Tuberculostearic acid

Author

Hannah G. Blitzblau

Ginkgo Bioworks

Novogy Inc.

Andrew L. Consiglio

Novogy Inc.

Ginkgo Bioworks

Paulo Teixeira

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Donald V. Crabtree

Novogy Inc.

Shuyan Chen

Ginkgo Bioworks

Novogy Inc.

Oliver Konzock

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Gamuchirai Chifamba

Novogy Inc.

Ginkgo Bioworks

Austin Su

Novogy Inc.

Annapurna Kamineni

Ginkgo Bioworks

Novogy Inc.

Kyle MacEwen

Novogy Inc.

Ginkgo Bioworks

Maureen Hamilton

Ginkgo Bioworks

Novogy Inc.

Vasiliki Tsakraklides

Novogy Inc.

Ginkgo Bioworks

Jens B Nielsen

Novo Nordisk Foundation Center for Biosustainability

BioInnovation Institute

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

Verena Siewers

Novo Nordisk Foundation Center for Biosustainability

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

A. Joe Shaw

Manus Biosynthesis

Novogy Inc.

Biotechnology for Biofuels

1754-6834 (eISSN)

Vol. 14 1 12

Subject Categories

Biochemistry and Molecular Biology

Biocatalysis and Enzyme Technology

Organic Chemistry

DOI

10.1186/s13068-020-01863-0

PubMed

33413611

More information

Latest update

1/19/2021