氮源及其浓度对三角褐指藻生长和脂肪酸组成的影响
EFFECTS OF NITROGEN SOURCES AND CONCENTRATIONS ON THE GROWTH AND FATTY ACID COMPOSITION OF PHAEODA-CTYLUM TRICORNUTUM
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摘要: 为了研究氮源的类型和浓度对微藻脂肪酸组成的影响,用含有不同浓度NO3-、NH4+、NH2CONH2的培养基,对三角褐指藻(Phaeodactylum tricornutum)进行了培养,并测定了其生长和脂肪酸组成.结果表明,培养基中不添加氮源时,三角褐指藻生长缓慢,但多不饱和脂肪酸(PUFAs)和C18脂肪酸(C18∶0,C18∶2(n-6),C18∶3(n-6))占总脂肪酸的比例较高;氮浓度较低(4+为氮源,生长较快;氮浓度较高(>3.5mmol/L)时,以NH2CONH2为氮源,生长较快.以NH4+或NH2CONH2为氮源时,EPA(Eicosapentaenoic acid)和PUFAs占总脂肪酸的比例随着其浓度的增加而上升;而以NO3-为氮源时,EPA和PUFAs随着NO3-浓度增加先上升后下降,最适NO3-浓度为1.8mmol/L,此时的EPA占总脂肪酸的比例为16.7%.EPA占干重(w/w)的比例,不管是哪种氮源,均随着氮浓度的增加而升高,但是在0.9-3.5mmol/L之间,3种氮源间EPA含量差异不显著.当氮源浓度为7.0mmol/L时,以NH2CONH2为氮源,EPA和PUFAs含量最高,分别为2.6%和4.4%.PUFAs占干重的比例随着NO3-浓度增加而下降,随NH2CONH2浓度增加而升高,而受NH4+浓度变化的影响不显著.Abstract: In order to investigate the effects of nitrogen sources and the concentrations on fatty acid composition in microalgae,a marine diatom,Phaeodactylum tricornutum,was cultured in batch with varied nitrogen sources(NO3-,NH4+,NH2CONH2)and at different concentrations.Its growth and fatty acid composition were investigated.The results showed that P.tricornutum grew poorly in the cultures without addition of any nitrogen source,however,polyunsaturated fatty acids(PUFAs)and C18 fatty acids(C18∶0,C18∶2(n-6),C18∶3(n-6))accounted for higher percentages of total fatty acids compared to those with addition of the nitrogen sources.At low concentrations of the nitrogen sources(4+,and grew faster with NH2CONH2 at high concentrations (>3.5mmol/L).With NH4+ or NH2CONH2 as nitrogen sources,EPA(eicosapentaenoic acid)and PUFAs in percentages of the total fatty acids increased when nitrogen concentrations were raised from 0.9 to 7.0mmol/L.With NO3-,the EPA and PUFAs percentages increased to the maximum at 1.8mmol/L NO3-,then decreased with the increased NO3- levels,while the highest EPA Was 16.7% of the total fatty acids.EPA content on a dry mass base(w/w)increased with the increasing nitrogen concentrations within a range of 0.9 to 7.0mmol/L regardless of the nitrogen sources.At 7.0mmol/L NH2CONH2,the alga achieved the highest contents(2.6%EPA and 4.4%PUFAs of the dry weight,respectively).PUFAs content on dry mass decreased with increased.NO3-,increased with enhanced NH2CONH2,but showed small variation with added NH4+.
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Keywords:
- Nitrogen sources /
- Phaeodactylum tricornutum /
- Growth /
- Fatty acids /
- PUFAs /
- EPA
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Science,2002,26(5):48-51.[梁英,麦康森,孙世春等.硝酸钠浓度对三角褐指藻(Phaeodactylum tricornutum)MACC/B226生长及脂肪酸组成的影响.海洋科学,2002,26(5):48-51][12] Wei D,Zhang X C,Sui H Z,et al.Effects of nitrogen sources and N/P ratio on cell growth,total lipid content and fatty a cid composition of Nannochloropsis oculata[J].Marine Scienc,2000,24(7):46-50.[魏东,张学成,隋正红,等.氮源和N/P对眼点拟微球藻的生长、总脂含量和脂肪酸组成的影响.海洋科学,2000,24(7):46-50.][13] Xu N J,Zhang X C,Fan X,et al.Effects of nitrogen source and concentration on growth rate and fatty acid composition of Ellipsoidion sp.(Eustigmatophyta)[J].J Appl Phycol.2001.13:463-469[14] Chor K T,Michael R T.Screening of diatoms for heterotrophic eicosapentaenoic acid production[J].J Appl Phycol,1996,8:59-64[15] Graeme A D,Volkman J K,Stephane M B,et al.Essential polyunsaturated fatty acids from 14 species of diatom(Bacillariophyceae)[J].Phytochemistry,1994,35(1):155-161[16] Chu W L,Phang S M,Goh S H.Environmental effects on growth and biochemical composition of Nitzschia inconspicua Grunow[J].J Appl Phycol,1996,8:389-396[17] Zhukova,Aizdaicher N A.Fatty acid compositon of 15 species of marine microalgae.Phytochemistry,1995,39(2):351-356[18] Liang Y,Mai K S,Sun S C.Total lipid and fatty acid composition of eight strains of marine diatoms[J].Chinese Journal of Oceanology and Limnology,2000,4(18):345-349[19] Yongmanitchai W,Ward P.Growth of and Omega-3 fatty acid production by Phaeodactylum tricornutum under different culture conditions[J].J Appl Envir Microbiol.1991,57(2):419-425[20] Goldberg I K,Chiara B,Cohen Z.Nitrogen starvation induces the accumulation of arachidonic acid in the freshwater green alga Parietochloris incisa(Trebuxiophyceae)[J].J Phycol,2002,38:991-994[21] Renaud S M,Parry D L,Thinh L V,et al.Effect of light intensity on the proximate biochemical and fatty acid composition of Isochrysis galbana and Nannochtorpsis oculata for use in tropical aquaculture[J].J Appl Phycol,1991,3:43-53[22] Yongmanitchai W,Ward P.Screening of algae for potential alternative sources of eicosapentaenoic acid.Phytochemistry,1991,30:2963-2967 Deyerberg J,Bang H O.Observations on populations in Greeland and Denmark,in nutritional evaluation of long-chain fatty acids[J].Lancet,1978,11:433-440[2] Wu K G,Chai X H.Protection of DHA dealing with the cerebrum and the eyesight[J].Food and Development,2000,21(2):41-43.[吴克刚,柴向华.DHA对大脑及视力的保健作用.食品研究与开发,2000,21(2):41-43][3] Jiang M X,Zheng Y Z.Total lipid and fatty acid composition of 14 species microalgae[J].Acta Hydrobiologica Sinica.2003,27(3):243-247.[蒋敏霞,郑亦周.14种海洋微藻总脂含量和脂肪酸组成研究.水生生物学报,2003,27(3):243-247][4] Yu J J,Li H F,Zhou H Q.Fatty acid composition of total,neutral and polar lipids of ten marine microalgae[J].Acta Hydrobiologica Sinica,1999,23(5):481-488.[俞建江,李荷芳,周汉秋.10种海洋微藻总脂、中性脂和极性脂的脂肪酸组成.水生生物学报,1999,23(5):481-488][5] Reitan K I,Rainuzzo J R,Olsen Y.Effect of nutrient limitation on fatty acid and lipid content of marine microalgae[J].J Phycol,1994,30:972-979[6] Liao Q B,Li W Q,Chen Q H.Effect of nutrition on the fatty acid composition in Phaeodactylum tricornutum[J].Marine Environmental Science,2000,19(2):6-9.[廖启斌,李文权,陈清花等.营养盐对三角褐指藻脂肪酸含量和百分组成的影响.海洋环境科学,2000,19(2):6-9][7] Grima E M,Sandez J A,Garcia J L,et al.EPA from Isochrysis galbana:growth conditions and productivity[J].Proces Biochem,1992,27:299-305[8] Yang X X,Yu H,Zeng X Q.Summarization of the factors affection thefatty acid composition of microalgae[J].Transactions of Oceanology and Limnology,2001,1:76-82.[杨秀霞于浩曾晓起.影响微藻脂肪酸组成因素概述.海洋湖沼通报,2001,1:76-82][9] Guillard R R L.Culture of phytoplankton for feeding marine invertebrates[A].Smith W L, Chanley M H.Culture of marine invertebrate animals[C],Plenum:New York Press,1975,29-60[10] Lepage G,Roy C.Improved recovery of fatty acids through direct transesterification without prior extraction or Purification[J].J Lipid Res,1984,25:1391-1396[11] Liang Y,Mai K S,Sun S C,et al.Effect of NaNO3 concentration on the growth and fatty acid compositions of Phaeodactylum tricornutum MACC/B226[J].Marine Science,2002,26(5):48-51.[梁英,麦康森,孙世春等.硝酸钠浓度对三角褐指藻(Phaeodactylum tricornutum)MACC/B226生长及脂肪酸组成的影响.海洋科学,2002,26(5):48-51][12] Wei D,Zhang X C,Sui H Z,et al.Effects of nitrogen sources and N/P ratio on cell growth,total lipid content and fatty a cid composition of Nannochloropsis oculata[J].Marine Scienc,2000,24(7):46-50.[魏东,张学成,隋正红,等.氮源和N/P对眼点拟微球藻的生长、总脂含量和脂肪酸组成的影响.海洋科学,2000,24(7):46-50.][13] Xu N J,Zhang X C,Fan X,et al.Effects of nitrogen source and concentration on growth rate and fatty acid composition of Ellipsoidion sp.(Eustigmatophyta)[J].J Appl Phycol.2001.13:463-469[14] Chor K T,Michael R T.Screening of diatoms for heterotrophic eicosapentaenoic acid production[J].J Appl Phycol,1996,8:59-64[15] Graeme A D,Volkman J K,Stephane M B,et al.Essential polyunsaturated fatty acids from 14 species of diatom(Bacillariophyceae)[J].Phytochemistry,1994,35(1):155-161[16] Chu W L,Phang S M,Goh S H.Environmental effects on growth and biochemical composition of Nitzschia inconspicua Grunow[J].J Appl Phycol,1996,8:389-396[17] Zhukova,Aizdaicher N A.Fatty acid compositon of 15 species of marine microalgae.Phytochemistry,1995,39(2):351-356[18] Liang Y,Mai K S,Sun S C.Total lipid and fatty acid composition of eight strains of marine diatoms[J].Chinese Journal of Oceanology and Limnology,2000,4(18):345-349[19] Yongmanitchai W,Ward P.Growth of and Omega-3 fatty acid production by Phaeodactylum tricornutum under different culture conditions[J].J Appl Envir Microbiol.1991,57(2):419-425[20] Goldberg I K,Chiara B,Cohen Z.Nitrogen starvation induces the accumulation of arachidonic acid in the freshwater green alga Parietochloris incisa(Trebuxiophyceae)[J].J Phycol,2002,38:991-994[21] Renaud S M,Parry D L,Thinh L V,et al.Effect of light intensity on the proximate biochemical and fatty acid composition of Isochrysis galbana and Nannochtorpsis oculata for use in tropical aquaculture[J].J Appl Phycol,1991,3:43-53[22] Yongmanitchai W,Ward P.Screening of algae for potential alternative sources of eicosapentaenoic acid.Phytochemistry,1991,30:2963-2967
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