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- Title
Production of 10-methyl branched fatty acids in yeast.
- Authors
Blitzblau, Hannah G.; Consiglio, Andrew L.; Teixeira, Paulo; Crabtree, Donald V.; Chen, Shuyan; Konzock, Oliver; Chifamba, Gamuchirai; Su, Austin; Kamineni, Annapurna; MacEwen, Kyle; Hamilton, Maureen; Tsakraklides, Vasiliki; Nielsen, Jens; Siewers, Verena; Shaw, A. Joe
- Abstract
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.
- Subjects
FATTY acids; UNSATURATED fatty acids; FREE fatty acids; DOUBLE bonds; YEAST; VEGETABLE oils
- Publication
Biotechnology for Biofuels, 2021, Vol 14, Issue 1, p1
- ISSN
1754-6834
- Publication type
Article
- DOI
10.1186/s13068-020-01863-0