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Triacylglycerol (TAG) metabolism is increasingly recognized not as a static competitive sink but as a dynamic and programmable carbon reservoir in microbiology. This forum recapitulates this paradigm shift and highlights reprogramming strategies to harness TAG cycling for replenishing precursors and/or energy required in the bioproduction of acetyl coenzyme A-derived compounds. Triacylglycerol (TAG) metabolism is increasingly recognized not as a static competitive sink but as a dynamic and programmable carbon reservoir in microbiology. This forum recapitulates this paradigm shift and highlights reprogramming strategies to harness TAG cycling for replenishing precursors and/or energy required in the bioproduction of acetyl coenzyme A-derived compounds.

The physiological functions of lysophosphatidylethanolamine (lysoPE) have not been fully elucidated. In this study, the effects of lysoPE on lipogenesis and lipolysis were investigated in a cultured human liver-derived cell line. The intracellular lipid profile was investigated in detail using liquid chromatography–tandem mass spectrometry (LC-MS/MS) to better understand the underlying mechanism. The expression of genes related to lipid metabolism and catabolism was analyzed using real-time PCR. LysoPE supplementation induced cellular lipid droplet formation and altered triacylglycerol (TAG) profiles. Furthermore, lysoPE downregulated expression of the TAG hydrolyzation regulation factor ATGL, and reduced the expression of fatty acid biosynthesis-related genes SREBP1 and SCD1. LC-MS/MS-based lipidomic profiling revealed that the addition of lysoPE 18:2 increased the PE species containing linoleic acyl, as well as the CE 18:2 species, likely due to the incorporation of linoleic acyl from lysoPE 18:2. Collectively, these findings suggest that lysoPE 18:2 is involved in lipid droplet formation by suppressing lipolysis and fatty acid biosynthesis. Thus, lysoPE might play a pathological role in the induction of fatty liver disease.

Purpose: The effects of intake of STGs containing esterified eicosapentaenoic acid (EPA) and medium-chain triglycerides (MCTs) on cardiorespiratory endurance have not yet been reported. This study aimed to examine the efficacy of interesterified structured lipids EPA and MCTs on cardiorespiratory endurance. Methods: This 8-week randomized double-blind placebo-controlled parallel-group study involved 19 healthy men. The participants were randomly assigned to a group that received interesterified structured lipids EPA and MCTs (STG group, 9 participants) or a group receiving a PM of EPA and MCTs (PM group, 10 participants). The outcome measures were time to exhaustion (TTE) and time to reach the anaerobic threshold in the peak oxygen uptake (VO2peak) test, VO2peak, and anaerobic threshold. Results: The increase in TTE in the VO2peak test after the intervention period compared with before the intervention period was significantly greater in the STG group (53 ± 53 s) than in the PM group (−10 ± 63 s; p < 0.05). Similarly, the increase in time to reach the anaerobic threshold was significantly greater in the STG group (82 ± 55 s) than in the PM group (−26 ± 52 s; p < 0.001). Conclusion: This study demonstrated that the consumption of interesterified structured lipids EPA and MCTs improved endurance in humans.

Similar to seeds, pollen tubes contain lipid droplets that store triacylglycerol (TAG), but the fate of this TAG as well as the enzymes involved in its breakdown are unknown. Therefore, two potential TAG lipases from tobacco and Arabidopsis, NtOBL1 (Oil body lipase 1) and AtOBL1, were investigated, especially with respect to their importance for pollen tube growth. We expressed NtOBL1 and AtOBL1 as fluorescent fusion proteins to study their localization by confocal microscopy. Furthermore, we overexpressed AtOBL1 in Nicotiana benthamiana leaves to characterize it enzymatically. The obl1 mutant was studied in respect to its pollen tube growth in vivo and its seed germination. Both NtOBL1 and AtOBL1 localized to lipid droplets. AtOBL1 was abundant in pollen tubes and seedlings, and acted as a lipase on TAG, diacylglycerol and 1-monoacylglycerol at a pH optimum of 5.5. The obl1 mutant was hampered in pollen tube growth, whereas seedling establishment was not affected under optimal conditions, even though AtOBL1 accounted for a major lipase activity in seeds. TAG could be a direct precursor for the synthesis of membrane lipids in pollen tubes and proteins of the OBL family involved in the flux of acyl groups.

Inhibition of autophagic flux prevented the synthesis of triacylglycerols, formation of lipid bodies, and degradation of ribosomal proteins RPS6 and RPL37 in nitrogen- or phosphate-starved Chlamydomonas cells. Abstract Autophagy is an intracellular catabolic process that allows cells to recycle unneeded or damaged material to maintain cellular homeostasis. This highly dynamic process is characterized by the formation of double-membrane vesicles called autophagosomes, which engulf and deliver the cargo to the vacuole. Flow of material through the autophagy pathway and its degradation in the vacuole is known as autophagic flux, and reflects the autophagic degradation activity. A number of assays have been developed to determine autophagic flux in yeasts, mammals, and plants, but it has not been examined yet in algae. Here we analyzed autophagic flux in the model green alga Chlamydomonas reinhardtii. By monitoring specific autophagy markers such as ATG8 lipidation and using immunofluorescence and electron microscopy techniques, we show that concanamycin A, a vacuolar ATPase inhibitor, blocks autophagic flux in Chlamydomonas. Our results revealed that vacuolar lytic function is needed for the synthesis of triacylglycerols and the formation of lipid bodies in nitrogen- or phosphate-starved cells. Moreover, we found that concanamycin A treatment prevented the degradation of ribosomal proteins RPS6 and RPL37 under nitrogen or phosphate deprivation. These results indicate that autophagy might play an important role in the regulation of lipid metabolism and the recycling of ribosomal proteins under nutrient limitation in Chlamydomonas.

The oleaginous yeast Lipomyces starkeyi is an excellent sustainable lipid producer, which can convert industrial wastes into lipids and accumulate triacylglycerols (TAG) by > 70% of its dry cell weight. Recent studies using omics technologies applied in L. starkeyi have aided in obtaining greater understanding of the important mechanisms of lipid metabolism in L. starkeyi. Therefore, the development of genetic engineering tools for L. starkeyi has led to accelerated efforts for a highly efficient production of lipids.This review focuses on the aspects of TAG and fatty acid synthesis pathways in L. starkeyi. We also present a quite effective strategy to obtain L. starkeyi mutants accumulating a larger amount of lipids and having a higher lipid production rate than the wild-type strain. The analysis of these mutants exhibiting high lipid production has led to the identification of important genes for achieving highly effective lipid production and thus advanced improvement in lipid production. Herein, our aim was to provide useful information to advance the development of L. starkeyi as a cost-effective TAG feedstock.Key Points•Oleaginous yeast Lipomyces starkeyi is an excellent sustainable lipid producer.•Efficient isolation of lipid-enriched L. starkeyi mutants depends on the low density of lipids.•Increased acyl-CoA synthesis pathway is important for improving lipid productivity.

In contrast to the consumption of long-chain triacylglycerols (LCT), consumption of medium- and long-chain triacylglycerols (MLCT) reduces the body fat and blood triacylglycerols (TAG) level in hypertriacylglycerolemic Chinese individuals. These responses may be affected by BMI because of obesity-induced insulin resistance. We aimed to compare the effects of consuming MLCT or LCT on reducing body fat and blood TAG level in hypertriacylglycerolemic Chinese subjects with different ranges of BMI. Employing a double-blind, randomized and controlled protocol, 101 hypertriacylglycerolemic subjects (including 67 men and 34 women) were randomly allocated to ingest 25-30 g/day MLCT or LCT oil as the only cooking oil for 8 consecutive weeks. Anthropometric measurements of body weight, BMI, body fat, WC, HC, blood biochemical variables, and subcutaneous fat area and visceral fat area in the abdomen were measured at week 0 and 8. As compared to subjects with BMI 24-28 kg/m² in the LCT group, corresponding subjects in the MLCT group showed significantly greater decrease in body weight, BMI, body fat, WC, ratio of WC to HC, total fat area and subcutaneous fat area in the abdomen, as well as blood TAG and LDL-C levels at week 8. Based upon our results, consumption of MLCT oil may reduce body weight, body fat, and blood TAG and LDL-C levels in overweight hypertriacylglycerolemic Chinese subjects but may not induce these changes in normal or obese hypertriacylglycerolemic subjects.

Triacylglycerol (TAG) and glycogen are the two major metabolites for carbon storage in most eukaryotic organisms. We investigated the glycogen metabolism of the oleaginous Y. lipolytica and found that this yeast accumulates up to 16% glycogen in its biomass. Assuming that elimination of glycogen synthesis would result in an improvement of lipid accumulation, we characterized and deleted the single gene coding for glycogen synthase, YlGSY1. The mutant was grown under lipogenic conditions with glucose and glycerol as substrates and we obtained up to 60% improvement in TAG accumulation compared to the wild type strain. Additionally, YlGSY1 was deleted in a background that was already engineered for high lipid accumulation. In this obese background, TAG accumulation was also further increased. The highest lipid content of 52% was found after 3 days of cultivation in nitrogen-limited glycerol medium. Furthermore, we constructed mutants of Y. lipolytica and S. cerevisiae that are deleted for both glycogen and TAG synthesis, demonstrating that the ability to store carbon is not essential. Overall, this work showed that glycogen synthesis is a competing pathway for TAG accumulation in oleaginous yeasts and that deletion of the glycogen synthase has beneficial effects on neutral lipid storage.

Microalgae are a valuable natural resource for a variety of value-added products. The growth of microalgae is determined by the impact of many factors, but, from the point of view of the implementation of autotrophic growth, light is of primary importance. This work presents an overview of the influence of light conditions on the growth of microalgae, the content of lipids, carotenoids, and the composition of fatty acids in their biomass, taking into account parameters such as the intensity, duration of lighting, and use of rays of different spectral composition. The optimal light intensity for the growth of microalgae lies in the following range: 26−400 µmol photons m−2 s−1. An increase in light intensity leads to an activation of lipid synthesis. For maximum lipid productivity, various microalgae species and strains need lighting of different intensities: from 60 to 700 µmol photons m−2 s−1. Strong light preferentially increases the triacylglyceride content. The intensity of lighting has a regulating effect on the synthesis of fatty acids, carotenoids, including β-carotene, lutein and astaxanthin. In intense lighting conditions, saturated fatty acids usually accumulate, as well as monounsaturated ones, and the number of polyunsaturated fatty acids decreases. Red as well as blue LED lighting improves the biomass productivity of microalgae of various taxonomic groups. Changing the duration of the photoperiod, the use of pulsed light can stimulate microalgae growth, the production of lipids, and carotenoids. The simultaneous use of light and other stresses contributes to a stronger effect on the productivity of algae.

• Microalgae accumulate high levels of oil under stress, but the underlying biosynthetic pathways are not fully understood. We sought to identify key regulators of lipid metabolism under stress conditions. • We found that the Chlamydomonas reinhardtii gene encoding the MYB-type transcription factor MYB1 is highly induced under stress conditions. Two myb1 mutants accumulated less total fatty acids and storage lipids than their parental strain upon nitrogen (N) depletion. • Transcriptome analysis revealed that genes involved in lipid metabolism are highly enriched in the wild-type but not in the myb1-1 mutant after 4 h of N depletion. Among these genes were several involved in the transport of fatty acids from the chloroplast to the endoplasmic reticulum (ER): acyl-ACP thioesterase (FAT1), Fatty Acid EXporters (FAX1, FAX2), and long-chain acyl-CoA synthetase1 (LACS1). Furthermore, overexpression of FAT1 in the chloroplast increased lipid production. These results suggest that, upon N depletion, MYB1 promotes lipid accumulation by facilitating fatty acid transport from the chloroplast to the ER. • This study identifies MYB1 as an important positive regulator of lipid accumulation in C. reinhardtii upon N depletion, adding another player to the established regulators of this process, including NITROGEN RESPONSE REGULATOR 1 (NRR1) and TRIACYLGLYCEROL ACCUMULATION REGULATOR 1 (TAR1).