Triacylglycerols(triglycerides,TAGs)are the major carbon and energy storage forms in various organisms,and important components of cellular membranes and signaling molecules;they have essential functions in multiple p...Triacylglycerols(triglycerides,TAGs)are the major carbon and energy storage forms in various organisms,and important components of cellular membranes and signaling molecules;they have essential functions in multiple physiological processes and stress regulation.Acyl-CoA:diacylglycerol acyltransferase(DGAT)catalyzes the final and only committed acylation step in the synthesis of TAGs in eukaryotes.The present work identified and isolated a novel gene,UpDGAT1,from the green tide alga Ulva prolifera.The activity of UpDGAT1 was confirmed by heterologous expression in a Saccharomyces cerevisiae TAG-deficient quadruple mutant.Results of thin-layer chromatography and BODIPY staining indicated that UpDGAT1 was able to restore TAG synthesis and lipid body formation in the yeast.Lipid analysis of yeast cells revealed that UpDGAT1 showed broad substrate specificity,accepting saturated as well as mono-and polyunsaturated acyl-CoAs as substrates.High salinity and high temperature stresses increased UpDGAT1 expression and TAG accumulation in U.prolifera.The present study provides clues to the functions of UpDGAT1 in TAG accumulation in,and stress adaptation of,U.prolifera.展开更多
Microalgal oils, depending on their degree of unsaturation, can be utilized as either nutritional supplements or fuels; thus, a feedstock with genetically designed and tunable degree of unsaturation is desirable to ma...Microalgal oils, depending on their degree of unsaturation, can be utilized as either nutritional supplements or fuels; thus, a feedstock with genetically designed and tunable degree of unsaturation is desirable to maximize process efficiency and product versatility. Systematic profiling of ex vivo (in yeast), in vitro, and in vivo activities of type-2 diacylglycerol acyltransferases in Nannochloropsis oceanica (NoDGAT2s or NoDGTTs), via reverse genetics, revealed that NoDGAT2A prefers saturated fatty acids (SFAs), NoDGAT2D prefers monounsaturated fatty acids (MUFAs), and NoDGAT2C exhibits the strongest activity toward polyunsaturated fatty acids (PUFAs). As NoDGAT2A, 2C, and 2D originated from the green alga, red alga, and eukaryotic host ancestral participants of secondary endosymbiosis, respectively, a mecha- nistic model of oleaginousness was unveiled, in which the indigenous and adopted NoDGAT2s formulated functional complementarity and specific transcript abundance ratio that underlie a rigid SFA:MUFA:PUFA hierarchy in triacylglycerol (TAG). By rationally modulating the ratio of NoDGAT2A':2C^D transcripts, a bank of N. oceanica strains optimized for nutritional supplement or fuel production with a wide range of degree of unsaturation were created, in which proportion of SFAs, MUFAs, and PUFAs in TAG varied by 1.3-, 3.7-, and 11.2-fold, respectively. This established a novel strategy to simultaneously improve productivity and quality of oils from industrial microalgae.展开更多
Two cDNAs encoding putative type 1 acyl-CoA: diacylglycerol acyltransferases (DGAT1, EC 2.3.1.20), were cloned from Tetraena mongolica Maxim., an extreme xerophyte with high oil content in the stems. The 1,488-bp a...Two cDNAs encoding putative type 1 acyl-CoA: diacylglycerol acyltransferases (DGAT1, EC 2.3.1.20), were cloned from Tetraena mongolica Maxim., an extreme xerophyte with high oil content in the stems. The 1,488-bp and 1,485-bp of the open reading frame (ORF) of the two cDNAs, designated as TmDGAT1a and TmDGAT1b, were both predicted to encode proteins of 495 and 494 amino acids, respectively. Southern blot analysis revealed that TmDGAT1a and TmDGAT1b both had low copy numbers in the T. mongolica genome. In addition to ubiquitous expression with different intensity in different tissues, including stems, leaves and roots, TmDGAT1a and TmDGAT1b, were found to be strongly induced by high salinity, drought and osmotic stress, resulting in a remarkable increase of triacylglycerol (TAG) accumulation in T. mongolica plantlets. TmDGAT1a and TmDGAT1b activities were confirmed in the yeast H1246 quadruple mutant (DGA1, LRO1, ARE1, ARE2) by restoring DGAT activity of the mutant host to produce TAG. Overexpression of TmDGAT1a and TmDGAT1b in soybean hairy roots as well as in T. mongolica calli both resulted in an increase in oil content (ranging from 37% to 108%), accompanied by altered fatty acid profiles.展开更多
Objective To identify the active compounds from the barks of Betula platyphylla for inhibitory on diacylglycerol acyltransferase(DGAT1).Methods Bioassay-guided fractionation resulted in the isolation of DGAT1 inhibi...Objective To identify the active compounds from the barks of Betula platyphylla for inhibitory on diacylglycerol acyltransferase(DGAT1).Methods Bioassay-guided fractionation resulted in the isolation of DGAT1 inhibitory activity of lupane triterpenes.Results Ten compounds were identified as lupenone(1),lupeol(2),betulinic acid(3),betulinaldehyde(4),betulin(5),3-deoxybetulonic acid(6),glochidonol(7),lup-20/29-ene-1β/3β-diol(8),3α-hydroxy-lup-20(29)-en-23,28-dioic acid(9),and 3α,11α-dihydroxy-23-oxo-lup-20(29)-en-28-oic acid(10).Compounds 3-6,9,and 10 inhibited DGAT1 with IC50 values ranging from(11.2±0.3)to(38.6±1.2)μmol/L.Conclusion Compounds 6,9,and 10 are first isolated from the barks of B.platyphylla.,and compounds 3-6,9,and 10 from the barks of B.platyphylla are responsible for the inhibition on DGAT1.展开更多
The enzymes of the acyl-coenzyme A:cholesterol acyltransferase(ACAT)family are responsible for the in vivo synthesis of neutral lipids.They are potential drug targets for the intervention of atherosclerosis,hyperlipid...The enzymes of the acyl-coenzyme A:cholesterol acyltransferase(ACAT)family are responsible for the in vivo synthesis of neutral lipids.They are potential drug targets for the intervention of atherosclerosis,hyperlipidemia,obesity,type II diabetes and even Alzheimer’s disease.ACAT family enzymes are integral endoplasmic reticulum(ER)membrane proteins and can be divided into ACAT branch and acyl-coenzyme A:diacylglycerol acyltransferase 1(DGAT1)branch according to their substrate specificity.The ACAT branch catalyzes synthesis of cholesteryl esters using long-chain fatty acyl-coenzyme A and cholesterol as substrates,while the DGAT1 branch catalyzes synthesis of triacylglycerols using fatty acylcoenzyme A and diacylglycerol as substrates.In this review,we mainly focus on the recent progress in the structural research of ACAT family enzymes,including their disulfide linkage,membrane topology,subunit interaction and catalysis mechanism.展开更多
基金Foundation item:The National Key Research and Development Program of China under contract No.2016YFC1402102the Central Public-interest Scientific Institution Basal Research Fund,CAFS under contract Nos 2020TD19 and 2020TD27+4 种基金the Major Scientific and Technological Innovation Project of Shandong Provincial Key Research and Development Program under contract 2019JZZY020706the International Exchange and Cooperation in Agriculture,Ministry of Agriculture and Rural Affairs of China-Science,Technology and Innovation Cooperation in Aquaculture with Tropical Countries along the Belt and RoadChina Agriculture Research System under contract No.CARS-50the Taishan Scholars Funding and Talent Projects of Distinguished Scientific Scholars in Agriculturethe National Ten Thousand Youth Talents Plan of 2014 under contract No.W02070268.
文摘Triacylglycerols(triglycerides,TAGs)are the major carbon and energy storage forms in various organisms,and important components of cellular membranes and signaling molecules;they have essential functions in multiple physiological processes and stress regulation.Acyl-CoA:diacylglycerol acyltransferase(DGAT)catalyzes the final and only committed acylation step in the synthesis of TAGs in eukaryotes.The present work identified and isolated a novel gene,UpDGAT1,from the green tide alga Ulva prolifera.The activity of UpDGAT1 was confirmed by heterologous expression in a Saccharomyces cerevisiae TAG-deficient quadruple mutant.Results of thin-layer chromatography and BODIPY staining indicated that UpDGAT1 was able to restore TAG synthesis and lipid body formation in the yeast.Lipid analysis of yeast cells revealed that UpDGAT1 showed broad substrate specificity,accepting saturated as well as mono-and polyunsaturated acyl-CoAs as substrates.High salinity and high temperature stresses increased UpDGAT1 expression and TAG accumulation in U.prolifera.The present study provides clues to the functions of UpDGAT1 in TAG accumulation in,and stress adaptation of,U.prolifera.
文摘Microalgal oils, depending on their degree of unsaturation, can be utilized as either nutritional supplements or fuels; thus, a feedstock with genetically designed and tunable degree of unsaturation is desirable to maximize process efficiency and product versatility. Systematic profiling of ex vivo (in yeast), in vitro, and in vivo activities of type-2 diacylglycerol acyltransferases in Nannochloropsis oceanica (NoDGAT2s or NoDGTTs), via reverse genetics, revealed that NoDGAT2A prefers saturated fatty acids (SFAs), NoDGAT2D prefers monounsaturated fatty acids (MUFAs), and NoDGAT2C exhibits the strongest activity toward polyunsaturated fatty acids (PUFAs). As NoDGAT2A, 2C, and 2D originated from the green alga, red alga, and eukaryotic host ancestral participants of secondary endosymbiosis, respectively, a mecha- nistic model of oleaginousness was unveiled, in which the indigenous and adopted NoDGAT2s formulated functional complementarity and specific transcript abundance ratio that underlie a rigid SFA:MUFA:PUFA hierarchy in triacylglycerol (TAG). By rationally modulating the ratio of NoDGAT2A':2C^D transcripts, a bank of N. oceanica strains optimized for nutritional supplement or fuel production with a wide range of degree of unsaturation were created, in which proportion of SFAs, MUFAs, and PUFAs in TAG varied by 1.3-, 3.7-, and 11.2-fold, respectively. This established a novel strategy to simultaneously improve productivity and quality of oils from industrial microalgae.
基金supported by the National Natural Science Foundation of China (30770224)the National Basic Research Program of China (2011CBA00901)
文摘Two cDNAs encoding putative type 1 acyl-CoA: diacylglycerol acyltransferases (DGAT1, EC 2.3.1.20), were cloned from Tetraena mongolica Maxim., an extreme xerophyte with high oil content in the stems. The 1,488-bp and 1,485-bp of the open reading frame (ORF) of the two cDNAs, designated as TmDGAT1a and TmDGAT1b, were both predicted to encode proteins of 495 and 494 amino acids, respectively. Southern blot analysis revealed that TmDGAT1a and TmDGAT1b both had low copy numbers in the T. mongolica genome. In addition to ubiquitous expression with different intensity in different tissues, including stems, leaves and roots, TmDGAT1a and TmDGAT1b, were found to be strongly induced by high salinity, drought and osmotic stress, resulting in a remarkable increase of triacylglycerol (TAG) accumulation in T. mongolica plantlets. TmDGAT1a and TmDGAT1b activities were confirmed in the yeast H1246 quadruple mutant (DGA1, LRO1, ARE1, ARE2) by restoring DGAT activity of the mutant host to produce TAG. Overexpression of TmDGAT1a and TmDGAT1b in soybean hairy roots as well as in T. mongolica calli both resulted in an increase in oil content (ranging from 37% to 108%), accompanied by altered fatty acid profiles.
基金Science and Technology Development Program of Jilin Province(201205099)
文摘Objective To identify the active compounds from the barks of Betula platyphylla for inhibitory on diacylglycerol acyltransferase(DGAT1).Methods Bioassay-guided fractionation resulted in the isolation of DGAT1 inhibitory activity of lupane triterpenes.Results Ten compounds were identified as lupenone(1),lupeol(2),betulinic acid(3),betulinaldehyde(4),betulin(5),3-deoxybetulonic acid(6),glochidonol(7),lup-20/29-ene-1β/3β-diol(8),3α-hydroxy-lup-20(29)-en-23,28-dioic acid(9),and 3α,11α-dihydroxy-23-oxo-lup-20(29)-en-28-oic acid(10).Compounds 3-6,9,and 10 inhibited DGAT1 with IC50 values ranging from(11.2±0.3)to(38.6±1.2)μmol/L.Conclusion Compounds 6,9,and 10 are first isolated from the barks of B.platyphylla.,and compounds 3-6,9,and 10 from the barks of B.platyphylla are responsible for the inhibition on DGAT1.
文摘The enzymes of the acyl-coenzyme A:cholesterol acyltransferase(ACAT)family are responsible for the in vivo synthesis of neutral lipids.They are potential drug targets for the intervention of atherosclerosis,hyperlipidemia,obesity,type II diabetes and even Alzheimer’s disease.ACAT family enzymes are integral endoplasmic reticulum(ER)membrane proteins and can be divided into ACAT branch and acyl-coenzyme A:diacylglycerol acyltransferase 1(DGAT1)branch according to their substrate specificity.The ACAT branch catalyzes synthesis of cholesteryl esters using long-chain fatty acyl-coenzyme A and cholesterol as substrates,while the DGAT1 branch catalyzes synthesis of triacylglycerols using fatty acylcoenzyme A and diacylglycerol as substrates.In this review,we mainly focus on the recent progress in the structural research of ACAT family enzymes,including their disulfide linkage,membrane topology,subunit interaction and catalysis mechanism.