Overproduction and secretion of free fatty acids through disrupted neutral lipid recycle in Saccharomyces cerevisiae

Leber, Christopher; Polson, Brian; Fernandez-Moya, Ruben; Silva, Nancy A. Da

doi:10.1016/j.ymben.2014.11.006

Cited by 79 publications

(67 citation statements)

References 39 publications

(66 reference statements)

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“…A similar phenomenon was also observed in Saccharomyces cerevisiae when native LACSs genes, faa1 and faa4 which account for the most of LACSs activity, were both removed [12]. Furthermore, FAs secretion can be greatly enhanced by additional expression of FAs synthesis genes in LACSs gene knockout strains [13, 14]. …”

Section: Introductionmentioning

confidence: 54%

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Characterization of long-chain acyl-CoA synthetases which stimulate secretion of fatty acids in green algae Chlamydomonas reinhardtii

Jia

Song

et al. 2016

Biotechnol Biofuels

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BackgroundMicroalgae biofuel has become the most promising renewable energy over the past few years. But limitations still exist because of its high cost. Although, efforts have been made in enhancement of lipid productivity, the major cost problem in harvesting and oil extraction is still intractable. Thus, the idea of fatty acids (FAs) secretion which can massively facilitate algae harvesting and oil extraction was investigated here.ResultsThe cDNAs of two long-chain acyl-CoA synthetases (LACSs) genes were cloned from Chlamydomonas reinhardtii and named as cracs1 and cracs2. They showed different substrate adaptation in the yeast complementation experiments. Cracs2 could utilize FAs C12:0, C14:0, C16:0, C18:0, C16:1 and C18:1, while crac1 could only utilize substrate C14:0, C16:1 and C18:1. Knockdown of cracs1 and cracs2 in C. reinhardtii resulted in accumulation of intracellular lipids. The total intracellular lipids contents of transgenic algae q-15 (knockdown of cracs1) and p-13 (knockdown of cracs2) were 45 and 55 %, respectively higher than that of cc849. Furthermore, FAs secretion was discovered in both transgenic algae. Secreted FAs can reach 8.19 and 9.66 mg/109 cells in q-15 and p-13, respectively.ConclusionThese results demonstrated the possibility of FAs secretion by microalgae and may give a new strategy of low-cost oil extraction. According to our findings, we proposed that FAs secretion may also be achieved in other species besides Chlamydomonas reinhardtii by knocking-down cracs genes, which may promote the future industrial application of microalgae biofuels.Electronic supplementary materialThe online version of this article (doi:10.1186/s13068-016-0598-7) contains supplementary material, which is available to authorized users.

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Section: Introductionmentioning

confidence: 54%

“…However, independent knockdown of LACS gene ( faa1 , faa2 , faa3 , faa4 , or fat1 ) showed little FAs secretion [7, 14]. We found single cracs knockdown was enough to induce FAs secretion in C. reinhardtii .…”

Section: Resultsmentioning

confidence: 89%

Characterization of long-chain acyl-CoA synthetases which stimulate secretion of fatty acids in green algae Chlamydomonas reinhardtii

Jia

Song

et al. 2016

Biotechnol Biofuels

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“…The overexpression of TesA thioesterase from E. coli or a truncated version of the Mus musculus ACOT5 thioesterase further increased FFA production up to 207 mg/L (Chen et al, 2014) and 520 mg/L (Runguphan and Keasling, 2014) respectively. A more recent study enhanced FFA production up to 2.2 g/L after multiple genetic engineering by disrupting acyl-CoA synthesis (Δfaa1 Δfaa4 Δfat1) and betaoxidation (Δfaa2, Δpxa1 Δpox1) together with enhancing neutral lipid formation and releasing FFA by an intracellular lipase (Leber et al, 2015). Nonetheless, no single study has been described so far in this regard in oleaginous organisms.…”

Section: Introductionmentioning

confidence: 99%

Combining metabolic engineering and process optimization to improve production and secretion of fatty acids

Ledesma-Amaro

Dulermo

Niehus

2016

Metabolic Engineering

150

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Microbial oils are sustainable alternatives to petroleum for the production of chemicals and fuels. Oleaginous yeasts are promising source of oils and Yarrowia lipolytica is the most studied and engineered one. Nonetheless the commercial production of biolipids is so far limited to high value products due to the elevated production and extraction costs. In order to contribute to overcoming these limitations we exploited the possibility of secreting lipids to the culture broth, uncoupling production and biomass formation and facilitating the extraction. We therefore considered two synthetic approaches, Strategy I where fatty acids are produced by enhancing the flux through neutral lipid formation, as typically occurs in eukaryotic systems and Strategy II where the bacterial system to produce free fatty acids is mimicked. The engineered strains, in a coupled fermentation and extraction process using alkanes, secreted the highest titer of lipids described so far, with a content of 120% of DCW.

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“…Study on the fatty acids secretion will provide rational strategies for the construction of robust strains with higher portion of extracellular fatty acids. Pioneering studies demonstrated that the acyl-CoA synthetases were involved in regulation of the production of extracellular fatty acids in S. cerevisiae, while many specific and unclear phenotypes were also observed [4][5][6][7][8]. (1) Fatty acids secretion was found in growth-phase dependent manner with the deletion of acyl-CoA synthetase genes [5].…”

Section: Introductionmentioning

confidence: 99%

“…Based on the deficiency of FAA1 and FAA4, higher titers of free fatty acids (207 and 520 mg/L) were further achieved by over-expressing the acyl-CoA thioesterase TesA and ACOTS, respectively [6]. Moreover, Leber et al reported that the deletions of acyl-CoA synthetases (FAA1, FAA4 and FAT1) genes and the ␤-oxidation pathway genes (FAA2, PXA1 and POX1) could lead to 1.3 g/L extracellular free fatty acids [4]. It was worth noting that the deletion of acyl-CoA synthetase genes could also lead to fatty acids secretion in many model microorganisms such as E. coli and cyanobacterium Synechocystis sp.…”

Section: Introductionmentioning

confidence: 99%

In depth understanding the molecular response to the enhanced secretion of fatty acids in S accharomyces cerevisiae due to one-step gene deletion of acyl-CoA synthetases

Fang

Dai

Zhao

et al. 2016

Process Biochemistry

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Overproduction and secretion of free fatty acids through disrupted neutral lipid recycle in Saccharomyces cerevisiae

Cited by 79 publications

References 39 publications

Characterization of long-chain acyl-CoA synthetases which stimulate secretion of fatty acids in green algae Chlamydomonas reinhardtii

Characterization of long-chain acyl-CoA synthetases which stimulate secretion of fatty acids in green algae Chlamydomonas reinhardtii

Combining metabolic engineering and process optimization to improve production and secretion of fatty acids

In depth understanding the molecular response to the enhanced secretion of fatty acids in S accharomyces cerevisiae due to one-step gene deletion of acyl-CoA synthetases

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