Production of 10-methyl branched fatty acids in yeast
Artikel i vetenskaplig tidskrift, 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.

10-Methylstearic acid

Tuberculostearic acid

Yarrowia lipolytica

Biobased lubricant

Författare

Hannah G. Blitzblau

Novogy Inc.

Ginkgo Bioworks

Andrew L. Consiglio

Novogy Inc.

Ginkgo Bioworks

Paulo Teixeira

Chalmers, Biologi och bioteknik, Systembiologi

Donald V. Crabtree

Novogy Inc.

Shuyan Chen

Ginkgo Bioworks

Novogy Inc.

Oliver Konzock

Chalmers, Biologi och bioteknik, Systembiologi

Gamuchirai Chifamba

Ginkgo Bioworks

Novogy Inc.

Austin Su

Novogy Inc.

Annapurna Kamineni

Ginkgo Bioworks

Novogy Inc.

Kyle MacEwen

Ginkgo Bioworks

Novogy Inc.

Maureen Hamilton

Novogy Inc.

Ginkgo Bioworks

Vasiliki Tsakraklides

Ginkgo Bioworks

Novogy Inc.

Jens B Nielsen

BioInnovation Institute

Chalmers, Biologi och bioteknik, Systembiologi

Novo Nordisk Fonden

Verena Siewers

Novo Nordisk Fonden

Chalmers, Biologi och bioteknik, Systembiologi

A. Joe Shaw

Novogy Inc.

Manus Biosynthesis

Biotechnology for Biofuels

17546834 (ISSN) 1754-6834 (eISSN)

Vol. 14 1 12

Ämneskategorier

Biokemi och molekylärbiologi

Biokatalys och enzymteknik

Organisk kemi

Infrastruktur

Chalmers infrastruktur för masspektrometri

DOI

10.1186/s13068-020-01863-0

PubMed

33413611

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Senast uppdaterat

2023-05-26