scholarly journals De novo Biosynthesis of Odd-Chain Fatty Acids in Yarrowia lipolytica Enabled by Modular Pathway Engineering

2020 ◽  
Vol 7 ◽  
Author(s):  
Young-kyoung Park ◽  
Rodrigo Ledesma-Amaro ◽  
Jean-Marc Nicaud
Keyword(s):  
Fatty Acids ◽  
Yarrowia Lipolytica ◽  
De Novo ◽  
Pathway Engineering ◽  
De Novo Biosynthesis ◽  
Odd Chain Fatty Acids ◽  
Chain Fatty Acids

scholarly journals Engineering precursor pools for increasing production of odd-chain fatty acids in Yarrowia lipolytica

2020 ◽  
Author(s):  
Young-Kyoung Park ◽  
Florence Bordes ◽  
Fabien Letisse ◽  
Jean-Marc Nicaud
Keyword(s):  
Fatty Acids ◽  
Yarrowia Lipolytica ◽  
Carbon Chain ◽  
Strain Engineering ◽  
Microbial Production ◽  
Total Lipids ◽  
Coa Transferase ◽  
Wide Range ◽  
Odd Chain Fatty Acids ◽  
Chain Fatty Acids

AbstractMicrobial production of lipids is one of the promising alternatives to fossil fuels with increasing environmental and energy concern. Odd-chain fatty acids (OCFA), a type of unusual lipids, are recently gaining a lot of interest as target compounds in microbial production due to their diverse applications in the medical, pharmaceutical, and chemical industries. In this study, we aimed to enhance the pool of precursors with three-carbon chain (propionyl-CoA) and five-carbon chain (β-ketovaleryl-CoA) for the production of OCFAs in Yarrowia lipolytica. We evaluated different propionate-activating enzymes and the overexpression of propionyl-CoA transferase gene from Ralstonia eutropha increased the accumulation of OCFAs by 3.8 times over control strain, indicating propionate activation is the limiting step of OCFAs synthesis. It was shown that acetate supplement was necessary to restore growth and to produce a higher OCFA contents in total lipids, suggesting the balance of the precursors between acetyl-CoA and propionyl-CoA is crucial for OCFA accumulation. To improve β-ketovaleryl-CoA pools for further increase of OCFA production, we co-expressed the bktB encoding β-ketothiolase in the producing strain, and the OCFA production was increased by 33 % compared to control. Combining strain engineering and the optimization of the C/N ratio promoted the OCFA production up to 1.87 g/L representing 62% of total lipids, the highest recombinant OCFAs titer reported in yeast, up to date. This study provides a strong basis for the microbial production of OCFAs and its derivatives having high potentials in a wide range of applications.

scholarly journals Engineering precursor pools for increasing production of odd-chain fatty acids in Yarrowia lipolytica

2021 ◽  
Vol 12 ◽  
pp. e00158
Author(s):  
Young-Kyoung Park ◽  
Florence Bordes ◽  
Fabien Letisse ◽  
Jean-Marc Nicaud
Keyword(s):  
Fatty Acids ◽  
Yarrowia Lipolytica ◽  
Odd Chain Fatty Acids ◽  
Chain Fatty Acids

Cloning of the Nocardia corallina polyhydroxyalkanoate synthase gene and production of poly-(3-hydroxybutyrate-co-3-hydroxyhexanoate) and poly-(3-hydroxyvalerate-co-3-hydroxyheptanoate)

1998 ◽  
Vol 44 (7) ◽  
pp. 687-691 ◽  
Author(s):  
Brian Hall ◽  
Jennifer Baldwin ◽  
Ho Gun Rhie ◽  
Douglas Dennis
Keyword(s):  
Fatty Acids ◽  
Ralstonia Eutropha ◽  
Carbon Sources ◽  
Pha Synthase ◽  
Broad Host Range ◽  
Coding Region ◽  
Reading Frame ◽  
Nocardia Corallina ◽  
Odd Chain Fatty Acids ◽  
Chain Fatty Acids

The polyhydroxyalkanoate (PHA) synthase gene (phaCNc) from Nocardia corallina was identified in a lambda library on a 6-kb BamHI fragment. A 2.8-kb XhoII subfragment was found to contain the ntact PHA synthase. This 2.8-kb fragment was subjected to DNA sequencing and was found to contain the coding region for the PHA synthase and a small downstream open reading frame of unknown function. On the basis of DNA sequence, phaCNc is closest in homology to the PHA synthases (phaCPaI and phaCPaII) of Pseudomonas aeruginosa (approximately 41% identity and 55% similarity). The 2.8-kb XhoII fragment containing phaCNc was subcloned into broad host range mobilizable plasmids and transferred into Escherichia coli, Klebsiella aerogenes (both containing a plasmid bearing phaA and phaB from Ralstonia eutropha), and PHA-negative strains of R. eutropha and Pseudomonas putida. The recombinant strains were grown on various carbon sources and the resulting polymers were analyzed. In these strains, the PHA synthase from N. corallina was able to mediate the production of poly(3-hydroxybutyrate-co-3-hydroxy-hexanoate) containing high levels of 3-hydroxyhexanoate when grown on hexanoate and larger even-chain fatty acids and poly(3-hydroxyvalerate-co-3-hydroxyheptanoate) containing high levels of 3-hydroxyheptanoate when grown on heptanoate or larger odd-chain fatty acids. Key words: polyhydroxyalkanoates (PHAs), Nocardia corallina, biodegradable, polyester.

scholarly journals Inclusion of Soluble Fiber in the Gestation Diet Changes the Gut Microbiota, Affects Plasma Propionate and Odd-Chain Fatty Acids Levels, and Improves Insulin Sensitivity in Sows

2020 ◽  
Vol 21 (2) ◽  
pp. 635 ◽  
Author(s):  
Chuanhui Xu ◽  
Chuanshang Cheng ◽  
Xiu Zhang ◽  
Jian Peng
Keyword(s):  
Fatty Acids ◽  
Insulin Sensitivity ◽  
Gut Microbiota ◽  
Systemic Inflammation ◽  
Guar Gum ◽  
Dietary Treatment ◽  
Perinatal Period ◽  
Soluble Fiber ◽  
Odd Chain Fatty Acids ◽  
Chain Fatty Acids

The transition from pregnancy to lactation is characterized by a progressive decrease in insulin sensitivity. Propionate increases with dietary fiber consumption and has been shown to improve insulin sensitivity. Recent studies suggest that plasma odd-chain fatty acids [OCFAs; pentadecanoic acid (C15:0) and heptadecanoic acid (C17:0)] that inversely correlated with insulin resistance are synthesized endogenously from gut-derived propionate. The present study investigated the effects of soluble fiber during gestation on gut microbiota, plasma non-esterified fatty acids and insulin sensitivity in sows. Sows were allocated to either control or 2.0% guar gum plus pregelatinized waxy maize starch (SF) dietary treatment during gestation. The SF addition changes the structure and composition of gut microbiota in sows. Genus Eubacterium increased by SF addition may promote intestinal propionate production. Moreover, the dietary SF increased circulating levels of plasma OCFAs, especially C17:0. The SF-fed sows had a higher insulin sensitivity and a lower systemic inflammation level during perinatal period. Furthermore, the plasma C15:0 and C17:0 was negatively correlated with the area under curve of plasma glucose after meal and plasma interleukin-6. In conclusion, dietary SF improves insulin sensitivity and alleviates systemic inflammation in perinatal sows, potentially related to its stimulating effect on propionate and OCFAs production.

scholarly journals Parasitic wasps do not lack lipogenesis

2021 ◽  
Author(s):  
Joachim Ruther ◽  
Lorena Prager ◽  
Tamara Pokorny
Keyword(s):  
Fatty Acids ◽  
De Novo ◽  
Model Organism ◽  
Parasitic Wasps ◽  
Primary Metabolites ◽  
De Novo Biosynthesis ◽  
Fat Reserves ◽  
Lipid Mass ◽  
Unlimited Access ◽  
Eleven Species

Fatty acids are crucial primary metabolites for virtually any creature on earth. Therefore, most organisms do not rely exclusively on nutritional supply with fatty acids but have the ability to synthesize fatty acids and triacylglycerides de novo from carbohydrates, a process called lipogenesis. The ubiquity of lipogenesis has been questioned by a series of studies reporting that many parasitic wasps (parasitoids) do not accumulate lipid mass despite having unlimited access to sugar. This has been interpreted as an evolutionary metabolic trait loss in parasitoids. Here, we demonstrate de novo biosynthesis of fatty acids from 13C-labeled α-D-glucose in eleven species of parasitoids from six families. We furthermore show with the model organism Nasonia vitripennis that lipogenesis occurs even when lipid reserves are still intact, but relative 13C-incorporation rates increase in females with widely depleted fat reserves. Therefore, we conclude that the presumed "lack of lipogenesis" in parasitoids needs to be re-evaluated.

scholarly journals Locations and contributions of the phosphotransferases EPT1 and CEPT1 to the biosynthesis of ethanolamine phospholipids

2020 ◽  
Vol 61 (8) ◽  
pp. 1221-1231 ◽  
Author(s):  
Yasuhiro Horibata ◽  
Hiromi Ando ◽  
Hiroyuki Sugimoto
Keyword(s):  
Fatty Acids ◽  
De Novo ◽  
Brefeldin A ◽  
Hek293 Cells ◽  
Intracellular Location ◽  
De Novo Biosynthesis ◽  
Substrate Preferences ◽  
Ethanolamine Phosphate ◽  
Immunohistochemical Analyses

The final step of the CDP-ethanolamine pathway is catalyzed by ethanolamine phosphotransferase 1 (EPT1) and choline/EPT1 (CEPT1). These enzymes are likely involved in the transfer of ethanolamine phosphate from CDP-ethanolamine to lipid acceptors such as 1,2-diacylglycerol (DAG) for PE production and 1-alkyl-2-acyl-glycerol (AAG) for the generation of 1-alkyl-2-acyl-glycerophosphoethanolamine. Here, we investigated the intracellular location and contribution to ethanolamine phospholipid (EP) biosynthesis of EPT1 and CEPT1 in HEK293 cells. Immunohistochemical analyses revealed that EPT1 localizes to the Golgi apparatus and CEPT1 to the ER. We created EPT1-, CEPT1-, and EPTI-CEPT1-deficient cells, and labeling of these cells with radio- or deuterium-labeled ethanolamine disclosed that EPT1 is more important for the de novo biosynthesis of 1-alkenyl-2-acyl-glycerophosphoethanolamine than is CEPT1. EPT1 also contributed to the synthesis of PE species containing the fatty acids 36:1, 36:4, 38:5, 38:4, 38:3, 40:6, 40:5, and 40:4. In contrast, CEPT1 was important for PE formation from shorter fatty acids such as 32:2, 32:1, 34:2, and 34:1. Brefeldin A treatment did not significantly affect the levels of the different PE species, indicating that the subcellular localization of the two enzymes is not responsible for their substrate preferences. In vitro enzymatic analysis revealed that EPT1 prefers AAG 16–20:4 > DAG 18:0–20:4 > DAG 16:0–18:1 = AAG 16–18:1 as lipid acceptors and that CEPT1 greatly prefers DAG 16:0–18:1 to other acceptors. These results suggest that EPT1 and CEPT1 differ in organelle location and are responsible for the biosynthesis of distinct EP species.

Regulation Of The Fatty Acid Composition Of Platelet Phospholipids

1981 ◽  
Author(s):  
M L McKean ◽  
J B Smith ◽  
M J Silver
Keyword(s):  
Fatty Acids ◽  
Fatty Acid Composition ◽  
Fatty Acid ◽  
Acid Composition ◽  
Unsaturated Fatty Acids ◽  
De Novo ◽  
Membrane Phospholipids ◽  
De Novo Biosynthesis ◽  
Coa Transferase

The fatty acid composition of cell membrane phospholipids does not remain constant after de novo biosynthesis, but undergoes continual remodelling. One of the major routes for remodelling probably includes the deacylation-reacylation steps of the Lands Pathway. This has been shown to be important for the incorporation of long chain, polyunsaturated fatty acids into phospholipids by liver and brain. An understanding of the mechanisms involved in these processes in platelets is especially important in light of the large stores of arachidonic acid (AA) in platelet phospholipids and the role of AA in hemostasis and thrombosis. Previous results from this laboratory have shown that the turnover of radioactive AA, 8,11,14-eicosatrienoic and 5,8,11,14,17-eicosapentaenoic acids in the phospholipids of resting platelets is more rapid than the turnover of radioactive C16 and C18 saturated and unsaturated fatty acids. However, little is known about how fatty acids, especially AA and its homologues, are incorporated into platelet phospholipids during de novo biosynthesis or how they are exchanged during remodelling.At least three enzymes are involved in the deacylation- reacylation of phospholipids: phospholipase A2; acyl CoA synthetase; and acyl CoA transferase. We have studied acyl CoA transferase and have found considerable activity in human platelet membranes. Experiments are in progress to determine the substrate specificity and other properties of this enzyme.

Odd-chain fatty acids as markers of the microbial colonisation of freshly-ingested grass and microbial contamination of dacron bag residues

2002 ◽  
Vol 2002 ◽  
pp. 50-50 ◽  
Author(s):  
E. J. Kim ◽  
R. T. Evans ◽  
J. K. S. Tweed ◽  
D. R. Davies ◽  
R. J. Merry ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Microbial Contamination ◽  
Technical Problems ◽  
Rumen Microbes ◽  
Rumen Degradation ◽  
Plant Enzymes ◽  
Odd Chain Fatty Acids ◽  
Chain Fatty Acids

The overall objective of our work is to assess the relative contributions of plant enzymes and rumen microbes to rumen degradation of freshly-ingested herbage. In situ techniques have been used extensively to compare rumen degradation characteristics of feeds, though there remain technical problems associated with microbial contamination of residues after incubation. We hypothesised that techniques to study microbial contamination might also provide insights into microbial colonisation. Our earlier studies (Lee et al., 1999) identified distinctive odd-chain fatty acids that could be used as microbial markers. A dacron bag study was conducted to examine the influence of dacron bag rinsing techniques on DM disappearance and microbial contamination in residues from fresh grass, assessed using odd-chain fatty acids as markers.

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