Production of 10-methyl branched fatty acids in yeast
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 t...
| Main Authors: | , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2021-01-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13068-020-01863-0 |
| _version_ | 1817977181162176512 |
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| author | Hannah G. Blitzblau Andrew L. Consiglio Paulo Teixeira Donald V. Crabtree Shuyan Chen Oliver Konzock Gamuchirai Chifamba Austin Su Annapurna Kamineni Kyle MacEwen Maureen Hamilton Vasiliki Tsakraklides Jens Nielsen Verena Siewers A. Joe Shaw |
| author_facet | Hannah G. Blitzblau Andrew L. Consiglio Paulo Teixeira Donald V. Crabtree Shuyan Chen Oliver Konzock Gamuchirai Chifamba Austin Su Annapurna Kamineni Kyle MacEwen Maureen Hamilton Vasiliki Tsakraklides Jens Nielsen Verena Siewers A. Joe Shaw |
| author_sort | Hannah G. Blitzblau |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-13T22:12:46Z |
| format | Article |
| id | doaj.art-594a94eec1eb421c801b204dc815a133 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-04-13T22:12:46Z |
| publishDate | 2021-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-594a94eec1eb421c801b204dc815a1332022-12-22T02:27:40ZengBMCBiotechnology for Biofuels1754-68342021-01-0114111710.1186/s13068-020-01863-0Production of 10-methyl branched fatty acids in yeastHannah G. Blitzblau0Andrew L. Consiglio1Paulo Teixeira2Donald V. Crabtree3Shuyan Chen4Oliver Konzock5Gamuchirai Chifamba6Austin Su7Annapurna Kamineni8Kyle MacEwen9Maureen Hamilton10Vasiliki Tsakraklides11Jens Nielsen12Verena Siewers13A. Joe Shaw14Novogy, Inc.Novogy, Inc.Department of Biology and Biological Engineering, Chalmers University of TechnologyNovogy, Inc.Novogy, Inc.Department of Biology and Biological Engineering, Chalmers University of TechnologyNovogy, Inc.Novogy, Inc.Novogy, Inc.Novogy, Inc.Novogy, Inc.Novogy, Inc.Department of Biology and Biological Engineering, Chalmers University of TechnologyDepartment of Biology and Biological Engineering, Chalmers University of TechnologyNovogy, Inc.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.https://doi.org/10.1186/s13068-020-01863-0Yarrowia lipolytica10-Methylstearic acidTuberculostearic acidBiobased lubricant |
| spellingShingle | Hannah G. Blitzblau Andrew L. Consiglio Paulo Teixeira Donald V. Crabtree Shuyan Chen Oliver Konzock Gamuchirai Chifamba Austin Su Annapurna Kamineni Kyle MacEwen Maureen Hamilton Vasiliki Tsakraklides Jens Nielsen Verena Siewers A. Joe Shaw Production of 10-methyl branched fatty acids in yeast Biotechnology for Biofuels Yarrowia lipolytica 10-Methylstearic acid Tuberculostearic acid Biobased lubricant |
| title | Production of 10-methyl branched fatty acids in yeast |
| title_full | Production of 10-methyl branched fatty acids in yeast |
| title_fullStr | Production of 10-methyl branched fatty acids in yeast |
| title_full_unstemmed | Production of 10-methyl branched fatty acids in yeast |
| title_short | Production of 10-methyl branched fatty acids in yeast |
| title_sort | production of 10 methyl branched fatty acids in yeast |
| topic | Yarrowia lipolytica 10-Methylstearic acid Tuberculostearic acid Biobased lubricant |
| url | https://doi.org/10.1186/s13068-020-01863-0 |
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