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https://www.readbyqxmd.com/read/28448543/heterologous-expression-of-a-thermophilic-diacylglycerol-acyltransferase-triggers-triglyceride-accumulation-in-escherichia-coli
#1
Beatriz Lázaro, Juan A Villa, Omar Santín, Matilde Cabezas, Cintia D F Milagre, Fernando de la Cruz, Gabriel Moncalián
Triglycerides (TAGs), the major storage molecules of metabolic energy and source of fatty acids, are produced as single cell oil by some oleogenic microorganisms. However, these microorganisms require strict culture conditions, show low carbon source flexibilities, lack efficient genetic modification tools and in some cases pose safety concerns. TAGs have essential applications such as behaving as a source for added-value fatty acids or giving rise to the production of biodiesel. Hence, new alternative methods are urgently required for obtaining these oils...
2017: PloS One
https://www.readbyqxmd.com/read/28328322/novel-lc-ms-ms-and-high-throughput-mass-spectrometric-assays-for-monoacylglycerol-acyltransferase-inhibitors
#2
Jenson Qi, John A Masucci, Wensheng Lang, Margery A Connelly, Gary W Caldwell, Ioanna Petrounia, Jennifer Kirkpatrick, Alexander N Barnakov, Geoffrey Struble, Robyn Miller, Keli Dzordzorine, Gee-Hong Kuo, Michael Gaul, Alessandro Pocai, Seunghun Lee
Monoacylglycerol acyltransferase enzymes (MGAT1, MGAT2, and MGAT3) convert monoacylglycerol to diacylglycerol (DAG). MGAT1 and MGAT2 are both implicated in obesity-related metabolic diseases. Conventional MGAT enzyme assays use radioactive substrates, wherein the product of the MGAT-catalyzed reaction is usually resolved by time-consuming thin layer chromatography (TLC) analysis. Furthermore, microsomal membrane preparations typically contain endogenous diacylglycerol acyltransferase (DGAT) from the host cells, and these DGAT activities can further acylate DAG to form triglyceride (TG)...
April 2017: SLAS Discovery
https://www.readbyqxmd.com/read/28325847/specialized-diacylglycerol-acyltransferase-contributes-to-extreme-fatty-acid-content-of-cuphea-seed-oil
#3
Umidjon Iskandarov, Jillian E Silva, Hae Jin Kim, Mariette Andersson, Rebecca E Cahoon, Keithanne Mockaitis, Edgar B Cahoon
Seed oils of many Cuphea sp. contain >90% of medium-chain fatty acids, such as decanoic acid (10:0). These seed oils, which are among the most compositionally variant in the plant kingdom, arise from specialized fatty acid biosynthetic enzymes and specialized acyltransferases. These include lysophosphatidic acid acyltransferases (LPAT) and diacylglycerol acyltransferases (DGAT) that are required for successive acylation of medium-chain fatty acids in the sn-2 and sn-3 positions of seed triacylglycerols (TAGs)...
March 21, 2017: Plant Physiology
https://www.readbyqxmd.com/read/28274201/analysis-of-triglyceride-synthesis-unveils-a-green-algal-soluble-diacylglycerol-acyltransferase-and-provides-clues-to-potential-enzymatic-components-of-the-chloroplast-pathway
#4
Carolina Bagnato, María B Prados, Gisela R Franchini, Natalia Scaglia, Silvia E Miranda, María V Beligni
BACKGROUND: Microalgal triglyceride (TAG) synthesis has attracted considerable attention. Particular emphasis has been put towards characterizing the algal homologs of the canonical rate-limiting enzymes, diacylglycerol acyltransferase (DGAT) and phospholipid:diacylglycerol acyltransferase (PDAT). Less work has been done to analyze homologs from a phylogenetic perspective. In this work, we used HMMER iterative profiling and phylogenetic and functional analyses to determine the number and sequence characteristics of algal DGAT and PDAT, as well as related sequences that constitute their corresponding superfamilies...
March 9, 2017: BMC Genomics
https://www.readbyqxmd.com/read/28247196/identification-and-characterization-of-a-novel-diacylglycerol-acyltransferase-gene-from-mortierella-alpina
#5
Xin-An Luo, Yuan-Min Zhu, Ting-Ting Liu, Xiao-Peng Wang, Peng-Peng Zhou, Zhen-Dong Bao, Long-Jiang Yu
OBJECTIVES: To clone and express a diacylglycerol acyltransferase (DGAT) gene from Mortierella alpina in Saccharomyces cerevisiae and characterize oil production and fatty acid composition of the resulting recombinant RESULTS: A new, full-length cDNA, putatively encoding a DGAT, was cloned from M. alpina. We subsequently cloned the gene, except the transmembrane-encoding region, termed MaDGAT, its molecular mass was 31.3 kDa. MaDGAT shares 75% identity with a DGAT from Mortierella verticillata NRRL 6337...
February 28, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28222675/identification-and-characterization-of-an-efficient-acyl-coa-diacylglycerol-acyltransferase-1-dgat1-gene-from-the-microalga-chlorella-ellipsoidea
#6
Xuejie Guo, Chengming Fan, Yuhong Chen, Jingqiao Wang, Weibo Yin, Richard R C Wang, Zanmin Hu
BACKGROUND: Oil in the form of triacylglycerols (TAGs) is quantitatively the most important storage form of energy for eukaryotic cells. Diacylglycerol acyltransferase (DGAT) is considered the rate-limiting enzyme for TAG accumulation. Chlorella, a unicellular eukaryotic green alga, has attracted much attention as a potential feedstock for renewable energy production. However, the function of DGAT1 in Chlorella has not been reported. RESULTS: A full-length cDNA encoding a putative diacylglycerol acyltransferase 1 (DGAT1, EC 2...
February 21, 2017: BMC Plant Biology
https://www.readbyqxmd.com/read/28168573/increasing-cocoa-butter-like-lipid-production-of-saccharomyces-cerevisiae-by-expression-of-selected-cocoa-genes
#7
Yongjun Wei, Michael Gossing, David Bergenholm, Verena Siewers, Jens Nielsen
Cocoa butter (CB) extracted from cocoa beans mainly consists of three different kinds of triacylglycerols (TAGs), 1,3-dipalmitoyl-2-oleoyl-glycerol (POP, C16:0-C18:1-C16:0), 1-palmitoyl-3-stearoyl-2-oleoyl-glycerol (POS, C16:0-C18:1-C18:0) and 1,3-distearoyl-2-oleoyl-glycerol (SOS, C18:0-C18:1-C18:0), but CB supply is limited. Therefore, CB-like lipids (CBL, which are composed of POP, POS and SOS) are in great demand. Saccharomyces cerevisiae produces TAGs as storage lipids, which are also mainly composed of C16 and C18 fatty acids...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28131831/response-of-genes-involved-in-lipid-metabolism-in-rat-epididymal-white-adipose-tissue-to-different-fasting-conditions-after-long-term-fructose-consumption
#8
Jin-Xiu Li, Da-Zhi Ke, Ling Yao, Shang Wang, Peng Ma, Li Liu, Guo-Wei Zuo, Li-Rong Jiang, Jian-Wei Wang
There has been much concern regarding the dietary fructose contributes to the development of metabolic syndrome. High-fructose diet changes the expression of genes involved in lipid metabolism. Levels of a number of hepatic lipogenic enzymes are increased by a high-carbohydrate diet in fasted-refed model rats/mice. Both the white adipose tissue (WAT) and the liver play a key role in the maintenance of nutrient homeostasis. Here, the aim of this study was to analyze the expression of key genes related to lipid metabolism in epididymal WAT (eWAT) in response to different fasting condition after long-term chronic fructose consumption...
January 25, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28103313/mycobacterium-marinum-degrades-both-triacylglycerols-and-phospholipids-from-its-dictyostelium-host-to-synthesise-its-own-triacylglycerols-and-generate-lipid-inclusions
#9
Caroline Barisch, Thierry Soldati
During a tuberculosis infection and inside lipid-laden foamy macrophages, fatty acids (FAs) and sterols are the major energy and carbon source for Mycobacterium tuberculosis. Mycobacteria can be found both inside a vacuole and the cytosol, but how this impacts their access to lipids is not well appreciated. Lipid droplets (LDs) store FAs in form of triacylglycerols (TAGs) and are energy reservoirs of prokaryotes and eukaryotes. Using the Dictyostelium discoideum/Mycobacterium marinum infection model we showed that M...
January 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28078675/overexpression-of-a-diacylglycerol-acyltransferase-gene-in-phaeodactylum-tricornutum-directs-carbon-towards-lipid-biosynthesis
#10
Jorge Dinamarca, Orly Levitan, G Kenchappa Kumaraswamy, Desmond S Lun, Paul G Falkowski
Under nutrient deplete conditions, diatoms accumulate between 15% to 25% of their dry weight as lipids, primarily as triacylglycerols (TAGs). As in most eukaryotes, these organisms produce TAGs via the acyl-CoA dependent Kennedy pathway. The last step in this pathway is catalyzed by diacylglycerol acyltransferase (DGAT) that acylates diacylglycerol (DAG) to produce TAG. To test our hypothesis that DGAT plays a major role in controlling the flux of carbon towards lipids, we overexpressed a specific type II DGAT gene, DGAT2D, in the model diatom Phaeodactylum tricornutum...
January 12, 2017: Journal of Phycology
https://www.readbyqxmd.com/read/28070221/nannochloropsis-a-rich-source-of-diacylglycerol-acyltransferases-for-engineering-of-triacylglycerol-content-in-different-hosts
#11
Krzysztof Zienkiewicz, Agnieszka Zienkiewicz, Eric Poliner, Zhi-Yan Du, Katharina Vollheyde, Cornelia Herrfurth, Sofia Marmon, Eva M Farré, Ivo Feussner, Christoph Benning
BACKGROUND: Photosynthetic microalgae are considered a viable and sustainable resource for biofuel feedstocks, because they can produce higher biomass per land area than plants and can be grown on non-arable land. Among many microalgae considered for biofuel production, Nannochloropsis oceanica (CCMP1779) is particularly promising, because following nutrient deprivation it produces very high amounts of triacylglycerols (TAG). The committed step in TAG synthesis is catalyzed by acyl-CoA:diacylglycerol acyltransferase (DGAT)...
2017: Biotechnology for Biofuels
https://www.readbyqxmd.com/read/27881664/lipid-disequilibrium-disrupts-er-proteostasis-by-impairing-erad-substrate-glycan-trimming-and-dislocation
#12
Milton To, Clark W H Peterson, Melissa A Roberts, Jessica L Counihan, Tiffany T Wu, Mercedes S Forster, Daniel K Nomura, James A Olzmann
The endoplasmic reticulum (ER) mediates the folding, maturation, and deployment of the secretory proteome. Proteins that fail to achieve their native conformation are retained in the ER and targeted for clearance by ER-associated degradation (ERAD), a sophisticated process that mediates the ubiquitin-dependent delivery of substrates to the 26S proteasome for proteolysis. Recent findings indicate that inhibition of long-chain acyl-CoA synthetases with triacsin C, a fatty acid analogue, impairs lipid droplet (LD) biogenesis and ERAD, suggesting a role for LDs in ERAD...
January 15, 2017: Molecular Biology of the Cell
https://www.readbyqxmd.com/read/27788579/a-diacylglycerol-transferase-1-inhibitor-is-a-potent-hepatitis-c-antiviral-in-vitro-but-not-in-patients-in-a-randomized-clinical-trial
#13
Edward Gane, Catherine Stedman, Kiran Dole, Jin Chen, Charles Daniel Meyers, Brigitte Wiedmann, Jin Zhang, Prakash Raman, Richard A Colvin
Hepatitis C virus (HCV) infection is a significant cause of liver disease affecting 80-150 million people globally. Diacylglycerol transferase 1 (DGAT-1), a triglyceride synthesis enzyme, is important for the HCV life cycle in vitro. Pradigastat, a potent DGAT-1 inhibitor found to lower triglycerides and HgbA1c in patients, was investigated for safety and efficacy in patients with HCV. This was a two-part study. In the in vitro study, the effect of pradigastat on virus production was evaluated in infected cells in culture...
February 10, 2017: ACS Infectious Diseases
https://www.readbyqxmd.com/read/27780240/expression-of-soluble-forms-of-yeast-diacylglycerol-acyltransferase-2-that-integrate-a-broad-range-of-saturated-fatty-acids-in-triacylglycerols
#14
Nawel Haïli, Julien Louap, Michel Canonge, Franjo Jagic, Christelle Louis-Mondésir, Thierry Chardot, Pierre Briozzo
The membrane proteins acyl-CoA:diacylglycerol acyltransferases (DGAT) are essential actors for triglycerides (TG) biosynthesis in eukaryotic organisms. Microbial production of TG is of interest for producing biofuel and value-added novel oils. In the oleaginous yeast Yarrowia lipolytica, Dga1p enzyme from the DGAT2 family plays a major role in TG biosynthesis. Producing recombinant DGAT enzymes pure and catalytically active is difficult, hampering their detailed functional characterization. In this report, we expressed in Escherichia coli and purified two soluble and active forms of Y...
2016: PloS One
https://www.readbyqxmd.com/read/27706370/diversity-and-evolution-of-plant-diacylglycerol-acyltransferase-dgats-unveiled-by-phylogenetic-gene-structure-and-expression-analyses
#15
Andreia Carina Turchetto-Zolet, Ana Paula Christoff, Franceli Rodrigues Kulcheski, Guilherme Loss-Morais, Rogerio Margis, Marcia Margis-Pinheiro
Since the first diacylglycerol acyltransferase (DGAT) gene was characterized in plants, a number of studies have focused on understanding the role of DGAT activity in plant triacylglycerol (TAG) biosynthesis. DGAT enzyme is essential in controlling TAGs synthesis and is encoded by different genes. DGAT1 and DGAT2 are the two major types of DGATs and have been well characterized in many plants. On the other hand, the DGAT3 and WS/DGAT have received less attention. In this study, we present the first general view of the presence of putative DGAT3 and WS/DGAT in several plant species and report on the diversity and evolution of these genes and its relationships with the two main DGAT genes (DGAT1 and DGAT2)...
October 2016: Genetics and Molecular Biology
https://www.readbyqxmd.com/read/27627966/tlr4-mutant-mice-are-resistant-to-acute-alcohol-induced-sterol-regulatory-element-binding-protein-activation-and-hepatic-lipid-accumulation
#16
Zhi-Hui Zhang, Xiao-Qian Liu, Cheng Zhang, Wei He, Hua Wang, Yuan-Hua Chen, Xiao-Jing Liu, Xi Chen, De-Xiang Xu
Previous studies demonstrated that acute alcohol intoxication caused hepatic lipid accumulation. The present study showed that acute alcohol intoxication caused hepatic lipid accumulation in Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic sterol-regulatory element binding protein (SREBP)-1, a transcription factor regulating fatty acid and triglyceride (TG) synthesis, was activated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice. Hepatic Fas, Acc, Scd-1 and Dgat-2, the key genes for fatty acid and TG synthesis, were up-regulated in alcohol-treated Tlr4-wild-type mice but not in Tlr4-mutant mice...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27578203/a-vernonia-diacylglycerol-acyltransferase-can-increase-renewable-oil-production
#17
Tomoko Hatanaka, William Serson, Runzhi Li, Paul Armstrong, Keshun Yu, Todd Pfeiffer, Xi-Le Li, David Hildebrand
Increasing the production of plant oils such as soybean oil as a renewable resource for food and fuel is valuable. Successful breeding for higher oil levels in soybean, however, usually results in reduced protein, a second valuable seed component. This study shows that by manipulating a highly active acyl-CoA:diacylglycerol acyltransferase (DGAT) the hydrocarbon flux to oil in oilseeds can be increased without reducing the protein component. Compared to other plant DGATs, a DGAT from Vernonia galamensis (VgDGAT1A) produces much higher oil synthesis and accumulation activity in yeast, insect cells, and soybean...
September 28, 2016: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/27546480/effect-of-quercetin-on-hepatitis-c-virus-life-cycle-from-viral-to-host-targets
#18
Ángela Rojas, Jose A Del Campo, Sophie Clement, Matthieu Lemasson, Marta García-Valdecasas, Antonio Gil-Gómez, Isidora Ranchal, Birke Bartosch, Juan D Bautista, Arielle R Rosenberg, Francesco Negro, Manuel Romero-Gómez
Quercetin is a natural flavonoid, which has been shown to have anti hepatitis C virus (HCV) properties. However, the exact mechanisms whereby quercetin impacts the HCV life cycle are not fully understood. We assessed the effect of quercetin on different steps of the HCV life cycle in Huh-7.5 cells and primary human hepatocytes (PHH) infected with HCVcc. In both cell types, quercetin significantly decreased i) the viral genome replication; ii) the production of infectious HCV particles and iii) the specific infectivity of the newly produced viral particles (by 85% and 92%, Huh7...
2016: Scientific Reports
https://www.readbyqxmd.com/read/27531967/diacylglycerol-acyltransferase-2-and-monoacylglycerol-acyltransferase-2-are-ubiquitinated-proteins-that-are-degraded-by-the-26s-proteasome
#19
Curtis Brandt, Pamela J McFie, Scot J Stone
Acyl-CoA:1,2-diacylglycerol acyltransferase (DGAT)-2 is one of the two DGAT enzymes that catalyzes the synthesis of triacylglycerol, which is an important form of stored energy for eukaryotic organisms. There is currently limited information available regarding how DGAT2 and triacylglycerol synthesis are regulated. Recent studies have indicated that DGAT2 can be regulated by changes in gene expression. How DGAT2 is regulated post-transcriptionally remains less clear. In this study, we demonstrated that DGAT2 is a very unstable protein and is rapidly degraded in an ubiquitin-dependent manner via the proteasome...
October 15, 2016: Biochemical Journal
https://www.readbyqxmd.com/read/27506614/overexpression-of-diacylglycerol-acyltransferase-in-yarrowia-lipolytica-affects-lipid-body-size-number-and-distribution
#20
Peter Gajdoš, Rodrigo Ledesma-Amaro, Jean-Marc Nicaud, Milan Čertík, Tristan Rossignol
In the oleaginous yeast Yarrowia lipolytica, the diacylglycerol acyltransferases (DGATs) are major factors for triacylglycerol (TAG) synthesis. The Q4 strain, in which the four acyltransferases have been deleted, is unable to accumulate lipids and to form lipid bodies (LBs). However, the expression of a single acyltransferase in this strain restores TAG accumulation and LB formation. Using this system, it becomes possible to characterize the activity and specificity of an individual DGAT. Here, we examined the effects of DGAT overexpression on lipid accumulation and LB formation in Y...
September 2016: FEMS Yeast Research
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