不同生长期威氏海链藻主要脂类成分分析
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摘要
采用超高效液相色谱-四极杆-飞行时间质谱联用分析系统(UPLC-Q-TOF MS),对威氏海链藻主要脂质的种类和含量进行研究,结果表明:该藻中主要脂类包括非极性脂三酰甘油(TAG),一类甜菜碱脂DGCC,一类磷脂磷脂酰胆碱(PC)以及4种光合膜脂,即二酰甘油单半乳糖脂(MGDG)、二酰甘油双半乳糖脂(DGDG)、二酰甘油硫代糖脂(SQDG)和磷脂酰甘油(PG)。所有TAG分子含量对数期最少,平台末期大量积累。从对数期到平台期,所有的PC含量减少,而DGCC恰好相反,只是到了平台末期有部分DGCC含量开始出现下降趋势。4种光合膜脂中,大多数SQDG、MGDG和PG在对数期到平台初期时含量不断增加,SQDG和MGDG从平台初期到平台末期含量出现降低的趋势,而PG含量继续增加;整个生长过程中,一种DGDG的含量不断降低。除了PG和SQDG外,其它脂类中都富含DHA和EPA多不饱和脂肪酸,具有较高的营养价值,可作为水产生物培养的重要生物饵料。
A marine microalgae, Thalassiosira weissflogii, which was respectively harvested at three selected time points(exponential phase, onset of stationary phase and end of stationary phase) by centrifugation at 5 000 r/min for 15 min and freeze-dried. Lipids of marine microalgae were extracted by the method of Bligh and Dyer. The qualitative and quantitative research on total lipids was analyzed by means of the flight time ultra-high performance liquid chromatogra phy tandem mass spectrometry(UPLC-Q-TOF-MS) using electron spraying ionization. Seven kinds of lipids were identified in T. weissflogii, which included neutral lipid TAG, betaine lipid DGCC, phospholipid PC and four classes of photosynthetic glycerolipid(MGDG, SQDG, DGDG, PG). All of the TAGs were low during the logarithmic growth phase but became higher in the stationary phase. The variation tendency of PC and DGCC were entirely different: a rapid increased of all DGCC levels was happened from the exponential phase to early stationary phase, but appears to decline in part from the period of early stationary phase to late stationary phase, meanwhile PC were all reduced to traces. As the culture proceeded from exponential phase to stationary phase, most of the MGDG, SQDG and PG increased, while the content of MGDG and SQDG reduced when T. weissflogii enters late stationary phase. On the other hand, an all decreasing trend was observed during the whole period of growth for DGDG(C20:5-16:2). Except for PG and SQDG, lipids were enriched with polyunsaturated fatty acids of EPA and DHA in Thalassiosira weissflogii which had high nutrient value. Therefore, T. weissflogii can use as an important food organisms in aquaculture.
A marine microalgae, Thalassiosira weissflogii, which was respectively harvested at three selected time points(exponential phase, onset of stationary phase and end of stationary phase) by centrifugation at 5 000 r/min for 15 min and freeze-dried. Lipids of marine microalgae were extracted by the method of Bligh and Dyer. The qualitative and quantitative research on total lipids was analyzed by means of the flight time ultra-high performance liquid chromatogra phy tandem mass spectrometry(UPLC-Q-TOF-MS) using electron spraying ionization. Seven kinds of lipids were identified in T. weissflogii, which included neutral lipid TAG, betaine lipid DGCC, phospholipid PC and four classes of photosynthetic glycerolipid(MGDG, SQDG, DGDG, PG). All of the TAGs were low during the logarithmic growth phase but became higher in the stationary phase. The variation tendency of PC and DGCC were entirely different: a rapid increased of all DGCC levels was happened from the exponential phase to early stationary phase, but appears to decline in part from the period of early stationary phase to late stationary phase, meanwhile PC were all reduced to traces. As the culture proceeded from exponential phase to stationary phase, most of the MGDG, SQDG and PG increased, while the content of MGDG and SQDG reduced when T. weissflogii enters late stationary phase. On the other hand, an all decreasing trend was observed during the whole period of growth for DGDG(C20:5-16:2). Except for PG and SQDG, lipids were enriched with polyunsaturated fatty acids of EPA and DHA in Thalassiosira weissflogii which had high nutrient value. Therefore, T. weissflogii can use as an important food organisms in aquaculture.
引文
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[12]González-Araya RL,Lebrun C,Quéré,et al.The selection of an ideal diet for Ostrea edulis(L.)broodstock conditioning(part B)[J].Aquaculture,2012,362(28):55-66.
[13]Rousch JM,Bingham SE,Sommerfeld MR.Changes in fatty acid profiles of thermo-intolerant and thermo-tolerant marine diatoms during temperature stress[J].Journal of Experiment Marine Biology and Ecology,2003,295,145-156.
[14]Bligh EG,Dyer WJ.A rapid method of total lipid extraction and purification[J].Canadian Journal of Biochemistry and Physiology,1959,37(8):911-917.
[15]Li S,Xu JL,Chen JJ,et al.Structural elucidation of co-eluted triglycerides in the marine diatom model organism Thalassiosira pseudonana by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry[J].Rapid Communications in Mass Spectrometry,2014,28(3):245-255.
[16]Hsu FF,Turk J.Structural characterization of triacylglycerols as lithiated adducts by electrospray ionization mass spectrometry using low-energy collisionally activated dissociation on a triple stage quadrupole instrument[J].Journal of the American Society for Mass Spectrometry,1999,10(7):587-599.
[17]Yan XJ,Li HY,Xu JL,et al.Analysis of phospholipids in microalga Nitzschia closterium by UPLC-Q-TOF-MS[J].Chinese Journal of Oceanology and Limnology,2010,28(1):106-112.
[18]Kato M,Sakai M,Adachi K,et al.Distribution of betaine lipids in marine algae[J].Phytochemistry,1996,42(5):1341-1345.
[19]Guella G,Frassanito R,Mancini I.A new solution for an old problem:the regiochemical distribution of the acyl chains in galactolipids can be established by electrospray ionization tandem mass spectrometry[J].Rapid Communications in Mass Spectrometry,2003,17(17):1982-1994.
[20]Xu JL,Chen DY,Yan XJ,et al.Global characterization of the photosynthetic glycerolipids from a marine diatom Stephanodiscus sp.by ultra performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight mass spectrometry[J].Analytica Chimica Acta,2010,663(1):60-68.
[21]Yan XJ,Chen DY,Xu JL,et al.Profiles of photosynthetic glycerolipids in three strains of Skeletonema determined by UPLC-Q-TOF-MS[J].Journal of Applied Phycology,2011,23(2):271-282.
[22]Frentzen M.Phosphatidylglycerol and sulfoquinovosyldiacylglycerol:Anionic membrane lipids and phosphate regulation[J].Current Opinion in Plant Biology,2004,7(3):270-276.
[23]Volkman JK,Jeffrey SW,Nichols PD,et al.Fatty acid and lipid composition of 10 species of microalgae used in mariculture[J].Journal of Experimental Marine Biology and Ecology,1989,128(3):219-240.
[24]Zhu CJ,Lee YK,Chao TM.Effect of temperature and growth phase on lipid and biochemical composition of Isochysis galbana TK1[J].Journal of Applied Phycology,1997,9(5):451-457.
[25]Li-Beisson Y,Shorrosh B,Beisson F,et al.Acyl-lipid metabolism[M]//The Arabidopsis Book.US:The American Society of Plant Biologist,2013:e0161.
[26]Gounaris K,Barber J.Monogalactosyldiacylglycerol-the most abundant polar lipid in nature[J].Trends in Biochemi cal Sciences,1983,8(10):378-381.
[27]Künzler K,Eichenberger W.Betaine lipids and zwitterionic phospholipids in plants and fungi[J].Phytochemistry,1997,46(5):883-892.
[28]Roberts LD,Mc Combie G,Titman CM,et al.A matter of fat:An introduction to lipidomic profiling methods[J].Journal of Chromatography B,2008,871(2):174-181.
[29]Vogel G,Eichenberger W.Betaine lipids in lower plants.Biosynthesis of DGTS and DGTA in Ochromonas danica(Chrysophyceae)and the possible role of DGTS in lipid metabolism[J].Plant and Cell Physiology,1992,33(4):427-436.
[30]Eichenberger W.Betaine lipids in lower plant.Distribution of DGTS,DGTA and phospholipids,and the intracellular localization and site of biosynthesis of DGTS[J].Plant physiology and Biochemistry,1993,31(2):213-221.
[31]吴克刚,柴向华.124n-3系多不饱和脂肪酸与健康[J].国外医学:卫生分册,2000,27(5):300-303.
[32]陈立侨,江洪波,周忠良,等.ω-3 HUFAs对中华绒鳌蟹幼体存活率及体脂肪酸组成的影响[J].水产学报,2000,24(5):448-452.
[33]林武杰.海洋微藻多不饱和脂肪酸研究进展[J].福建水产,2013,35(1):78-82.
[34]Kiatmetha P,Siangdang W,Bunnag B,et al.Enhancement of survival and metamorphosis rates of Penaeus monodon larvae by feeding with the diatom Thalassiosira weissflogii[J].Aquaculture International,2011,19(4):599-609.
[35]Glencross BD,Smith DM,Thomas MR,et al.The effects of dietary lipid amount and fatty-acid composition on the digestibility of lipids by the prawn Penaeus monodon[J].Aquaculture,2002,205(1):157-169.
[36]Chen M,Liu H,Chen B.Effects of dietary essential fatty acids on reproduction rates of a subtropical calanoid copepod,Acartia erythraea[J].Marine Ecology Progress Series,2012,455:95-110.
[37]Thompson P,Guo M,Harrison P.The influence of irradiance on the biochemical composition of three phytoplankton species and their nutritional value for larvae of the Pacific oyster(Crassostrea gigas)[J].Marine Biology,1993,117(2):259-268.
[2]Kumari P,Kumar M,Reddy CRK,et al.Algal lipids,fatty acids and sterols[M].Woodhead Publishing Limited:Cambridge,UK,2013:87-134.
[3]Zhao FJ,Xu JL,Chen JJ,et al.Structural elucidation of two types of novel glycosphingolipids in three strains of Skeletonema by liquid chromatography coupled with mass spectrometry[J].Rapid Communications in Mass Spectrometry,2013,27(3):1535-1547.
[4]Grima EM,Perez JA,Sanchez JL,et al.EPA from Isochrysis galbana.Growth conditions and productivity[J].Pro cess Biochemistry,1992,27(5):299-305.
[5]Yongmanitchai W,Ward OP.Growth of and omega-3 fatty acid production by Phaeodactylum tricornutumunder different culture conditions[J].Applied and Environmental Microbiology,1991,57(2):419-425.
[6]魏东,张学成.细胞生长时期对两种海洋微藻总脂含量和脂肪酸组成的影响[J].青岛海洋大学学报:自然科学版,2000,30(3):503-503.
[7]陈炜,梁明明,白永安,等.小球藻不同生长时期总脂含量和脂肪酸组成的变化[J].水产科学,2013,32(9):545-548.
[8]刘皓,高永利,殷克东,等.不同氮磷比对中肋骨条藻和威氏海链藻生长特性的影响[J].热带海洋学报,2010,29(6):92-97.
[9]Frost BW.Effects of size and concentration of food particles on the feeding behavior of the marine planktonic copepod Calanus pacificus[J].Limnol Oceanogr,1972,17(6):805-815.
[10]Downs JN,Lorenzen CJ.Carbon:Pheopigment ratios of zooplankton fecal pellets as an index of herbivorous feeding[J].Limnology and Oceanography,1985,30(5):1024-1036.
[11]Emmerson W.Ingestion,growth and development of Penaeus indicus larvae as a function of Thalassiosira weissflogii cell concentration[J].Marine Biology,1980,58(1):65-73.
[12]González-Araya RL,Lebrun C,Quéré,et al.The selection of an ideal diet for Ostrea edulis(L.)broodstock conditioning(part B)[J].Aquaculture,2012,362(28):55-66.
[13]Rousch JM,Bingham SE,Sommerfeld MR.Changes in fatty acid profiles of thermo-intolerant and thermo-tolerant marine diatoms during temperature stress[J].Journal of Experiment Marine Biology and Ecology,2003,295,145-156.
[14]Bligh EG,Dyer WJ.A rapid method of total lipid extraction and purification[J].Canadian Journal of Biochemistry and Physiology,1959,37(8):911-917.
[15]Li S,Xu JL,Chen JJ,et al.Structural elucidation of co-eluted triglycerides in the marine diatom model organism Thalassiosira pseudonana by ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry[J].Rapid Communications in Mass Spectrometry,2014,28(3):245-255.
[16]Hsu FF,Turk J.Structural characterization of triacylglycerols as lithiated adducts by electrospray ionization mass spectrometry using low-energy collisionally activated dissociation on a triple stage quadrupole instrument[J].Journal of the American Society for Mass Spectrometry,1999,10(7):587-599.
[17]Yan XJ,Li HY,Xu JL,et al.Analysis of phospholipids in microalga Nitzschia closterium by UPLC-Q-TOF-MS[J].Chinese Journal of Oceanology and Limnology,2010,28(1):106-112.
[18]Kato M,Sakai M,Adachi K,et al.Distribution of betaine lipids in marine algae[J].Phytochemistry,1996,42(5):1341-1345.
[19]Guella G,Frassanito R,Mancini I.A new solution for an old problem:the regiochemical distribution of the acyl chains in galactolipids can be established by electrospray ionization tandem mass spectrometry[J].Rapid Communications in Mass Spectrometry,2003,17(17):1982-1994.
[20]Xu JL,Chen DY,Yan XJ,et al.Global characterization of the photosynthetic glycerolipids from a marine diatom Stephanodiscus sp.by ultra performance liquid chromatography coupled with electrospray ionization-quadrupole-time of flight mass spectrometry[J].Analytica Chimica Acta,2010,663(1):60-68.
[21]Yan XJ,Chen DY,Xu JL,et al.Profiles of photosynthetic glycerolipids in three strains of Skeletonema determined by UPLC-Q-TOF-MS[J].Journal of Applied Phycology,2011,23(2):271-282.
[22]Frentzen M.Phosphatidylglycerol and sulfoquinovosyldiacylglycerol:Anionic membrane lipids and phosphate regulation[J].Current Opinion in Plant Biology,2004,7(3):270-276.
[23]Volkman JK,Jeffrey SW,Nichols PD,et al.Fatty acid and lipid composition of 10 species of microalgae used in mariculture[J].Journal of Experimental Marine Biology and Ecology,1989,128(3):219-240.
[24]Zhu CJ,Lee YK,Chao TM.Effect of temperature and growth phase on lipid and biochemical composition of Isochysis galbana TK1[J].Journal of Applied Phycology,1997,9(5):451-457.
[25]Li-Beisson Y,Shorrosh B,Beisson F,et al.Acyl-lipid metabolism[M]//The Arabidopsis Book.US:The American Society of Plant Biologist,2013:e0161.
[26]Gounaris K,Barber J.Monogalactosyldiacylglycerol-the most abundant polar lipid in nature[J].Trends in Biochemi cal Sciences,1983,8(10):378-381.
[27]Künzler K,Eichenberger W.Betaine lipids and zwitterionic phospholipids in plants and fungi[J].Phytochemistry,1997,46(5):883-892.
[28]Roberts LD,Mc Combie G,Titman CM,et al.A matter of fat:An introduction to lipidomic profiling methods[J].Journal of Chromatography B,2008,871(2):174-181.
[29]Vogel G,Eichenberger W.Betaine lipids in lower plants.Biosynthesis of DGTS and DGTA in Ochromonas danica(Chrysophyceae)and the possible role of DGTS in lipid metabolism[J].Plant and Cell Physiology,1992,33(4):427-436.
[30]Eichenberger W.Betaine lipids in lower plant.Distribution of DGTS,DGTA and phospholipids,and the intracellular localization and site of biosynthesis of DGTS[J].Plant physiology and Biochemistry,1993,31(2):213-221.
[31]吴克刚,柴向华.124n-3系多不饱和脂肪酸与健康[J].国外医学:卫生分册,2000,27(5):300-303.
[32]陈立侨,江洪波,周忠良,等.ω-3 HUFAs对中华绒鳌蟹幼体存活率及体脂肪酸组成的影响[J].水产学报,2000,24(5):448-452.
[33]林武杰.海洋微藻多不饱和脂肪酸研究进展[J].福建水产,2013,35(1):78-82.
[34]Kiatmetha P,Siangdang W,Bunnag B,et al.Enhancement of survival and metamorphosis rates of Penaeus monodon larvae by feeding with the diatom Thalassiosira weissflogii[J].Aquaculture International,2011,19(4):599-609.
[35]Glencross BD,Smith DM,Thomas MR,et al.The effects of dietary lipid amount and fatty-acid composition on the digestibility of lipids by the prawn Penaeus monodon[J].Aquaculture,2002,205(1):157-169.
[36]Chen M,Liu H,Chen B.Effects of dietary essential fatty acids on reproduction rates of a subtropical calanoid copepod,Acartia erythraea[J].Marine Ecology Progress Series,2012,455:95-110.
[37]Thompson P,Guo M,Harrison P.The influence of irradiance on the biochemical composition of three phytoplankton species and their nutritional value for larvae of the Pacific oyster(Crassostrea gigas)[J].Marine Biology,1993,117(2):259-268.