Production of free monounsaturated fatty acids by metabolically engineered Escherichia coli
Open accessAbstract
BACKGROUND: Monounsaturated fatty acids (MUFAs) are the best components for biodiesel when considering the low temperature fluidity and oxidative stability. However, biodiesel derived from vegetable oils or microbial lipids always consists of significant amounts of polyunsaturated and saturated fatty acids (SFAs) alkyl esters, which hampers its practical applications. Therefore, the fatty acid composition should be modified to increase MUFA contents as well as enhancing oil and lipid production. RESULTS: The model microorganism Escherichia coli was engineered to produce free MUFAs. The fatty acyl-ACP thioesterase (AtFatA) and fatty acid desaturase (SSI2) from Arabidopsis thaliana were heterologously expressed in E. coli BL21 star(DE3) to specifically release free unsaturated fatty acids (UFAs) and convert SFAs to UFAs. In addition, the endogenous fadD gene (encoding acyl-CoA synthetase) was disrupted to block fatty acid catabolism while the native acetyl-CoA carboxylase (ACCase) was overexpressed to increase the malonyl coenzyme A (malonyl-CoA) pool and boost fatty acid biosynthesis. The finally engineered strain BL21ΔfadD/pE-AtFatAssi2&pA-acc produced 82.6 mg/L free fatty acids (FFAs) under shake-flask conditions and FFAs yield on glucose reached about 3.3% of the theoretical yield. Two types of MUFAs, palmitoleate (16:1Δ9) and cis-vaccenate (18:1Δ11) made up more than 75% of the FFA profiles. Fed-batch fermentation of this strain further enhanced FFAs production to a titer of 1.27 g/L without affecting fatty acid compositions. CONCLUSIONS: This study demonstrated the possibility to regulate fatty acid composition by using metabolic engineering approaches. FFAs produced by the recombinant E. coli strain consisted of high-level MUFAs and biodiesel manufactured from these fatty acids would be more suitable for current diesel engines.
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SFA and MUFA in microalgae can be excellent biodiesel source due to high oxidative stability and lower NOx emissions than PUFA, though high SFA melting point can cause cold-weather gelling.
“SFA and MUFA in microalgae can be an excellent source of biodiesel due to their high oxidative stability and lesser NO X emissions in comparison to polyunsaturated fatty acids (PUFA), except that the high melting point of SFA can cause gelling of fuel in cold weather.[12,13].”
From Biomolecular transitions and lipid accumulation in green microalgae monitored by FTIR and Raman analysis (2020) · cites this paper as [13]
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- Authors
- Cao, Y., Liu, W., Xu, X., Zhang, H., Wang, J., & Xian, M.
- Affiliations
- Chinese Academy of Sciences (CN)
- Corresponding author
- Xian, Mo
- Venue
- Biotechnology for Biofuels
- Published
- 2014-04-10
- Volume
- 7
- Issue
- 1
- Pages
- 59-59
- ISSN
- 1754-6834 (ISSN-L)
- PMC
- PMC4021618
- OpenAlex
- W2129969521
- Cited by
- 107
- References
- 62
- Citations by year
- 2026: 4 · 2025: 7 · 2024: 9 · 2023: 16 · 2022: 12 · 2021: 10 · 2020: 5 · 2019: 7 · 2018: 8 · 2017: 9 · 2016: 12 · 2015: 4 · 2014: 4
- FWCI
- 3.97
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- Biotechnology for Biofuels (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Institutional knowledge base (Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences) (submittedVersion)
- Funders
- National Natural Science Foundation of China (2012BAD32B06, 31200030, 21202179, 12-1-4-9-(3)-jch, 21376255)
- Key Technologies Research and Development Program
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Keywords
- monounsaturated fatty acids, free fatty acids, biodiesel, Escherichia coli, fatty acid desaturase, malonyl-CoA, fatty acid composition, palmitoleic acid, fed-batch fermentation, fatty acid synthesis
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