scholarly journals Drop-In Biofuel production by using fatty acid photodecarboxylase from Chlorella variabilis in the oleaginous yeast Yarrowia lipolytica

2018 ◽  
Author(s):  
Stefan Bruder ◽  
Eva Johanna Moldenhauer ◽  
Robert Denis Lemke ◽  
Rodrigo Ledesma-Amaro ◽  
Johannes Kabisch
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Yarrowia Lipolytica ◽  
Light Exposure ◽  
Synthetic Medium ◽  
Biofuel Production ◽  
Fatty Aldehyde ◽  
Oleaginous Yeasts ◽  
Chlorella Variabilis ◽  
Microwell Plate

AbstractBackgroundOleaginous yeasts are potent hosts for the renewable production of lipids and harbor great potential for derived products, such as biofuels. Several promising processes have been described that produce hydrocarbon drop-in biofuels based on fatty acid decarboxylation and fatty aldehyde decarbonylation. Unfortunately, besides fatty aldehyde toxicity and high reactivity, the most investigated enzyme, aldehyde-deformylating oxygenase, shows unfavorable catalytic properties which hindered high yields in previous metabolic engineering approaches.ResultsTo demonstrate an alternative alkane production pathway for oleaginous yeasts, we describe the production of diesel-like, odd-chain alkanes and alkenes, by heterologously expressing a recently discovered light-driven oxidase from Chlorella variabilis (CvFAP) in Yarrowia lipolytica. Initial experiments showed that only strains engineered to have an increased pool of free fatty acids showed to be susceptible to sufficient decarboxylation. Providing these strains with glucose and light in a synthetic medium resulted in titers of 10.9 mg/L of hydrocarbons. Using custom 3D printed labware for lighting bioreactors, and an automated pulsed glycerol fed-batch strategy, intracellular titers of 58.7 mg/L were achieved.ConclusionsOleaginous yeasts such as Yarrowia lipolytica can transform renewable resources such as glycerol into fatty acids and lipids. By heterologously expressing a fatty acid photodecarboxylase from the algae Chlorella variabilis hydrocarbons were produced in several scales from microwell plate to 400 ml bioreactors. The developed bioprocess shows a route to the renewable production of hydrocarbons for a variety of applications ranging from representing a substrate for further enzymatic or chemical modification or as a drop-in biofuel blend.Short abstractOleaginous yeasts are potent hosts for the renewable production of lipids, fatty acids and derived products such as biofuels. Here, we describe, the production of odd-numbered alkanes and alkenes with a length of 17 and 15 carbons by expression of a fatty acid photodecarboxylase (CvFAP) from Chlorella variabilis in different Yarrowia lipolytica strains under different regimes of blue light exposure in several scales from microwell plate to 400 ml bioreactors.

scholarly journals Application of Random Mutagenesis and Synthetic FadR Promoter for de novo Production of ω-Hydroxy Fatty Acid in Yarrowia lipolytica

2021 ◽  
Vol 9 ◽  
Author(s):  
Beom Gi Park ◽  
Junyeob Kim ◽  
Eun-Jung Kim ◽  
Yechan Kim ◽  
Joonwon Kim ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Metabolic Engineering ◽  
Palmitic Acid ◽  
Yarrowia Lipolytica ◽  
Cell Sorting ◽  
De Novo ◽  
Random Mutagenesis ◽  
Oleaginous Yeasts ◽  
Synthetic Promoters ◽  
Base Strain

As a means to develop oleaginous biorefinery, Yarrowia lipolytica was utilized to produce ω-hydroxy palmitic acid from glucose using evolutionary metabolic engineering and synthetic FadR promoters for cytochrome P450 (CYP) expression. First, a base strain was constructed to produce free fatty acids (FFAs) from glucose using metabolic engineering strategies. Subsequently, through ethyl methanesulfonate (EMS)-induced random mutagenesis and fluorescence-activated cell sorting (FACS) screening, improved FFA overproducers were screened. Additionally, synthetic promoters containing bacterial FadR binding sequences for CYP expression were designed to respond to the surge of the concentration of FFAs to activate the ω-hydroxylating pathway, resulting in increased transcriptional activity by 14 times from the third day of culture compared to the first day. Then, endogenous alk5 was screened and expressed using the synthetic FadR promoter in the developed strain for the production of ω-hydroxy palmitic acid. By implementing the synthetic FadR promoter, cell growth and production phases could be efficiently decoupled. Finally, in batch fermentation, we demonstrated de novo production of 160 mg/L of ω-hydroxy palmitic acid using FmeN3-TR1-alk5 in nitrogen-limited media. This study presents an excellent example of the production of ω-hydroxy fatty acids using synthetic promoters with bacterial transcriptional regulator (i.e., FadR) binding sequences in oleaginous yeasts.

scholarly journals Preferential synthesis of very long chain polyunsaturated fatty acids in eutreptiella sp. (eugelnozoa) revealed by chromatographic and transcriptomic analyses

2020 ◽  
Author(s):  
Rita C. Kuo ◽  
Huan Zhang ◽  
James D. Stuart ◽  
Anthony A. Provatas ◽  
Linda Hannick ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Polyunsaturated Fatty Acids ◽  
Carbon Fixation ◽  
Acyl Carrier Protein ◽  
Lipid Production ◽  
Fatty Acid Desaturase ◽  
Biofuel Production ◽  
Type I ◽  
Long Chain

AbstractAlgal lipids are important fuel storage molecules in algae and a currency for energy transfer in the marine food chain as well as materials for biofuel production, but their production and regulation are not well understood in many species including the common coastal phytoplankton Eutreptiella spp. Here, using gas chromatography-tandem mass spectrometry (GC/MS/MS), we discovered 24 types of fatty acids (FAs) in Eutreptiella sp. with a relatively high proportion of long chain unsaturated FAs. The abundances of C16, C18 and saturated FAs decreased when phosphate in the culture medium was depleted. Among the 24 FAs, docosahexaenoic acid (22:6) and eicosapentaenoic acid (20:5) were the most abundant, suggesting that Eutreptiella sp. preferentially invests in the synthesis of very long chain polyunsaturated fatty acids (VLCPFA). Further transcriptomic analysis revealed that Eutreptiella sp. likely synthesizes VLCPFA via Δ8 pathway and uses type I and II fatty acid synthases. Using RT-qPCR, we found that some of the lipid production genes, such as β-ketoacyl-ACP reductase, fatty acid desaturase, acetyl-CoA carboxylase, acyl carrier protein, Δ8 desaturase, and Acyl-ACP thioesterase, were more actively expressed during light period. Besides, two carbon-fixation genes were more highly expressed in the high lipid illuminated cultures, suggesting a linkage between photosynthesis and lipid production.

scholarly journals A Brief Journey into the History of and Future Sources and Uses of Fatty Acids

2021 ◽  
Vol 8 ◽  
Author(s):  
Michela Cerone ◽  
Terry K. Smith
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Large Scale ◽  
Primary Source ◽  
Biofuel Production ◽  
Cell Factory ◽  
Primary Role ◽  
Animal Fats ◽  
Technological Advances ◽  
History Of

Fats and lipids have always had a primary role in the history of humankind, from ancient civilisations to the modern and contemporary time, going from domestic and cosmetic uses, to the first medical applications and later to the large-scale industrial uses for food, pharmaceutical, cosmetics, and biofuel production. Sources and uses of those have changed during time following the development of chemical sciences and industrial technological advances. Plants, fish, and animal fats have represented the primary source of lipids and fats for century. Nowadays, the use of fatty acid sources has taken a turn: industries are mainly interested in polyunsaturated fatty acids (PUFAs), which have beneficial properties in human health; and also, for high-value fatty acids product for innovative and green production of biofuel and feedstocks. Thus, the constant increase in demand of fatty acids, the fact that marine and vegetable sources are not adequate to meet the high level of fatty acids required worldwide and climate change, have determined the necessity of the search for renewable and sustainable sources for fatty acids. Biotechnological advances and bioengineering have started looking at the genetic modification of algae, bacteria, yeasts, seeds, and plants to develop cell factory able to produce high value fatty acid products in a renewable and sustainable manner. This innovative approach applied to FA industry is a peculiar example of how biotechnology can serve as a powerful mean to drive the production of high value fatty acid derivatives on the concept of circular bioeconomy, based on the reutilisation of organic resources for alternative and sustainable productive patterns that are environmentally friendly.

Production of Medium Chain Fatty Acids by Yarrowia lipolytica: Combining Molecular Design and TALEN to Engineer the Fatty Acid Synthase

2017 ◽  
Vol 6 (10) ◽  
pp. 1870-1879 ◽  
Author(s):  
Coraline Rigouin ◽  
Marc Gueroult ◽  
Christian Croux ◽  
Gwendoline Dubois ◽  
Vinciane Borsenberger ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Yarrowia Lipolytica ◽  
Fatty Acid Synthase ◽  
Molecular Design ◽  
Medium Chain ◽  
Medium Chain Fatty Acids ◽  
Chain Fatty Acids

scholarly journals Fatty acid composition as a parameter of using vegetable oils for biofuel production

2017 ◽  
Vol 169 (2) ◽  
pp. 38-42
Author(s):  
Marta AMBROSEWICZ-WALACIK ◽  
Małgorzata TAŃSKA ◽  
Marek WALACIK
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Acid Composition ◽  
Vegetable Oils ◽  
Raw Materials ◽  
Biofuel Production ◽  
Evening Primrose ◽  
Continuous Quality ◽  
Cold Pressed

The aim of the study was to show the validity of determining the quality of oils intendent for the biofuels production. One of the main discriminant that should be used to assess the suitability of oil raw materials for the fuel purposes is the composition of fatty acids. The research was conducted on 13 different samples of vegetable oils, that were cold-pressed from the seeds of various species, including rape, camelina, flax (light and dark), mustard, milk thistle, hemp, evening primrose oil, amaranth, sunflower, soybean, and cumin. The analysis of the fatty acid composition showed significant differences between the analysed oil samples, both within the species and varieties of oilseeds. The results indicate the need for continuous quality control of oils and fats led to the production of biodiesel in the small households and large commercial enterprises, because as it was indicated by the analysis of fatty acids composition rape seeds of different varieties are characterized by the diverse quality.

scholarly journals Herbal Industry Wastes as Potential Materials for Biofuel Production

Proceedings ◽  
2020 ◽  
Vol 51 (1) ◽  
pp. 6
Author(s):  
Aneta Sienkiewicz ◽  
Alicja Piotrowska-Niczyporuk ◽  
Andrzej Bajguz
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Unsaturated Fatty Acids ◽  
Methyl Esters ◽  
Saturated Fatty Acids ◽  
Research Work ◽  
Biodiesel Production ◽  
Biofuel Production ◽  
Gas Chromatography Mass Spectrometry ◽  
Ultrasound Assisted Extraction

Due to the increasing awareness of the depletion of fossil fuel resources and environmental issues, biodiesel as alternative fuel has become more and more attractive in the recent years. In this research, the characterization of herbal industry wastes as a potential feedstock for biodiesel production was carried out. There results of analytical identification of the fatty acid methyl esters (FAME) obtained in the transesterification reaction are presented. The reaction conditions were optimized, considering hexane ratio and catalyst concentration (methanol and KOH) for both steps. The FAME were extracted from the herbal samples by ultrasound-assisted extraction and subsequently were identified by gas chromatography-mass spectrometry (GC-MS) using single ion monitoring (SIM) method. Additionally, the selected properties of some bioesters were analyzed. This study determined the compounds which are ideal for fuel production. The unsaturated fatty acids were found in higher amounts than saturated fatty acids. Linoleic acid (C18:2n6c) was the major unsaturated fatty acid in herbal wastes, while palmitic acid (16:0) was the major saturated fatty acid. The application of the optimized method also revealed differences in the physical and chemical properties of isolated FAME mixtures compared to conventional diesel fuel. In this research work, for the first time, the possibilities of using the herbal industry wastes as a potential feedstock for biodiesel production are assessed.

scholarly journals Engineering Yarrowia lipolytica as a platform for synthesis of drop-in transportation fuels and oleochemicals

2016 ◽  
Vol 113 (39) ◽  
pp. 10848-10853 ◽  
Author(s):  
Peng Xu ◽  
Kangjian Qiao ◽  
Woo Suk Ahn ◽  
Gregory Stephanopoulos
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Free Fatty Acids ◽  
Chain Length ◽  
Yarrowia Lipolytica ◽  
Fatty Acid Synthase ◽  
Acyl Carrier Protein ◽  
Ethyl Esters ◽  
Pathway Engineering ◽  
Efficient Production

Harnessing lipogenic pathways and rewiring acyl-CoA and acyl-ACP (acyl carrier protein) metabolism in Yarrowia lipolytica hold great potential for cost-efficient production of diesel, gasoline-like fuels, and oleochemicals. Here we assessed various pathway engineering strategies in Y. lipolytica toward developing a yeast biorefinery platform for sustainable production of fuel-like molecules and oleochemicals. Specifically, acyl-CoA/acyl-ACP processing enzymes were targeted to the cytoplasm, peroxisome, or endoplasmic reticulum to generate fatty acid ethyl esters and fatty alkanes with tailored chain length. Activation of endogenous free fatty acids and the subsequent reduction of fatty acyl-CoAs enabled the efficient synthesis of fatty alcohols. Engineering a hybrid fatty acid synthase shifted the free fatty acids to a medium chain-length scale. Manipulation of alternative cytosolic acetyl-CoA pathways partially decoupled lipogenesis from nitrogen starvation and unleashed the lipogenic potential of Y. lipolytica. Taken together, the strategies reported here represent promising steps to develop a yeast biorefinery platform that potentially upgrades low-value carbons to high-value fuels and oleochemicals in a sustainable and environmentally friendly manner.

Platelet Aggregation in Finnish Men and Its Relation to Fatty Acids in Platelets, Plasma and Adipose Tissue

1985 ◽  
Vol 54 (03) ◽  
pp. 563-569 ◽  
Author(s):  
M K Salo ◽  
E Vartiainen ◽  
P Puska ◽  
T Nikkari
Keyword(s):  
Fatty Acids ◽  
Adipose Tissue ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Platelet Aggregation ◽  
Acid Composition ◽  
Saturated Fatty Acids ◽  
Free Living ◽  
Ω3 Fatty Acids ◽  
Induced Aggregation

SummaryPlatelet aggregation and its relation to fatty acid composition of platelets, plasma and adipose tissue was determined in 196 randomly selected, free-living, 40-49-year-old men in two regions of Finland (east and southwest) with a nearly twofold difference in the IHD rate.There were no significant east-southwest differences in platelet aggregation induced with ADP, thrombin or epinephrine. ADP-induced platelet secondary aggregation showed significant negative associations with all C20-C22 ω3-fatty acids in platelets (r = -0.26 - -0.40) and with the platelet 20: 5ω3/20: 4ω 6 and ω3/ ω6 ratios, but significant positive correlations with the contents of 18:2 in adipose tissue (r = 0.20) and plasma triglycerides (TG) (r = 0.29). Epinephrine-induced aggregation correlated negatively with 20: 5ω 3 in plasma cholesteryl esters (CE) (r = -0.23) and TG (r = -0.29), and positively with the total percentage of saturated fatty acids in platelets (r = 0.33), but had no significant correlations with any of the ω6-fatty acids. Thrombin-induced aggregation correlated negatively with the ω3/6ω ratio in adipose tissue (r = -0.25) and the 20: 3ω6/20: 4ω 6 ratio in plasma CE (r = -0.27) and free fatty acids (FFA) (r = -0.23), and positively with adipose tissue 18:2 (r = 0.23) and 20:4ω6 (r = 0.22) in plasma phospholipids (PL).The percentages of prostanoid precursors in platelet lipids, i. e. 20: 3ω 6, 20: 4ω 6 and 20 :5ω 3, correlated best with the same fatty acids in plasma CE (r = 0.32 - 0.77) and PL (r = 0.28 - 0.74). Platelet 20: 5ω 3 had highly significant negative correlations with the percentage of 18:2 in adipose tissue and all plasma lipid fractions (r = -0.35 - -0.44).These results suggest that, among a free-living population, relatively small changes in the fatty acid composition of plasma and platelets may be reflected in significant differences in platelet aggregation, and that an increase in linoleate-rich vegetable fat in the diet may not affect platelet function favourably unless it is accompanied by an adequate supply of ω3 fatty acids.

EFFECT OF GINGER, LEMURU FISH OIL SAPONIFICATION AND OLIVE OIL ON COMPOSITION FATTY ACID OF BEEF

2014 ◽  
Vol 4 (1) ◽  
pp. 31-39
Author(s):  
Siwitri Kadarsih
Keyword(s):  
Fatty Acids ◽  
Fatty Acid ◽  
Fish Oil ◽  
Olive Oil ◽  
Rice Bran ◽  
Dry Matter ◽  
Unsaturated Fatty Acids ◽  
Saturated Fatty Acids ◽  
Omega 3 ◽  
Omega 6

The objective was to get beef that contain unsaturated fatty acids (especially omega 3 and 6), so as to improve intelligence, physical health for those who consume. The study design using CRD with 3 treatments, each treatment used 4 Bali cattle aged approximately 1.5 years. Observations were made 8 weeks. Pasta mixed with ginger provided konsentrat. P1 (control); P2 (6% saponification lemuru fish oil, olive oil 1%; rice bran: 37.30%; corn: 62.70%; KLK: 7%, ginger paste: 100 g); P3 (lemuru fish oil saponification 8%, 2% olive oil; rice bran; 37.30; corn: 62.70%; KLK: 7%, ginger paste: 200 g). Konsentrat given in the morning as much as 1% of the weight of the cattle based on dry matter, while the grass given a minimum of 10% of the weight of livestock observation variables include: fatty acid composition of meat. Data the analyzies qualitative. The results of the study showed that the composition of saturated fatty acids in meat decreased and an increase in unsaturated fatty acids, namely linoleic acid (omega 6) and linolenic acid (omega 3), and deikosapenta deikosaheksa acid.Keywords : 

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