蔷薇藻活性产物的代谢调控及藻蓝蛋白的分离纯化
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摘要
蔷薇藻是一种属红藻门的海洋单细胞微藻,内含淀粉核,蔷薇藻在培养过程中能产生胞外多糖、藻胆蛋白和色素等生物活性物质。本文对蔷薇藻培养基中的氮源进行优化,分别以硝酸钾和尿素为唯一氮源,得到二者的最适浓度;分离纯化该藻的藻蓝蛋白,研究了该藻蓝蛋白的氨基酸组成、稳定性以及体外抗氧化活性。得到以下几个方面的结果:1、以硝酸钾为氮源,当含氮量在1.85～296mmol/L之间,蔷薇藻都能保持良好的生长;在最佳含氮量为3.7mmol/L时,生物量和藻蓝蛋白分别提高了5.13%和15.94%;含氮量为296mmol/L时,获得最高胞外多糖产量,提高了254.87%,表明硝酸盐是蔷薇藻的合适氮源。2、以尿素为氮源,含氮量在1.85～29.6mmol/L之间,藻细胞都能保持良好的生长;在最佳含氮量为14.8mmol/L时,生物量、胞外多糖和藻蓝蛋白分别提高了26.92%、23.58%和35.22%;但是当含氮量≥74mmol/L时,藻细胞死亡,表明高浓度的尿素能抑制蔷薇藻的生长甚至有致死作用。3、硝酸还原酶的酶活测定结果表明了该酶与藻的生物量成正相关,且硝酸钾比尿素更能激发该酶的活性。超氧化物歧化酶的活性随着含氮量的增大而升高。4、经两步硫酸铵沉淀和DEAE Sepharose Fast Flow离子交换柱层析后,得到蔷薇藻的纯藻蓝蛋白的纯度为3.92,SDS-PAGE表明该蛋白由两个亚基组成,分子量分别为18和22kDa,氨基酸测定结果表明该蛋白由17种氨基酸组成。5、藻蓝蛋白在pH为中性、低温避光下具有更高的稳定性。6、通过藻蓝蛋白的体外抗氧化实验,得出纯化后的藻蓝蛋白比粗藻蓝蛋白的抗氧化性更强,而且呈剂量依赖效应。
The unicellular red alga,Rhodella reticulata with a pyrenoid in it,can synthesize some bioactive substances such as exopolysaccharides,phycobiliprotein and pigments.In this paper,we investigated the optimization of nitrogen source,and made potassium nitrate and urea as a sole nitrogen source respectively,obtained the best nitrogen concentration,isolated and purified the phycocyanin from Rhodella reticulata,analyzed the amino acid composition and stability of the phycocyanin,as well as antioxidant activity of phycocyanin in vitro.The results were as follows:
1.Potassium nitrate as a nitrogen source,Rhodella reticulata grew well when nitrogen concentration was between 1.85～296mmol/L.The production of biomass and phycocyanin had increased 5.13%and 15.94%under the best nitrogen concentration(3.7mmol/L).When the nitrogen concentration was 296mmol/L,the maximum exopolysaccharides of Rhodella reticulata increased 254.87%.Therefore, potassium nitrate was the optimum nitrogen source.
2.Urea as a nitrogen source,Rhodella reticulata grew well when nitrogen concentration was between 1.85～29.6mmol/L,the production of biomass, exopolysaccharides and phycocyanin had increased 26.92%,23.58%and 35.22%at the best nitrogen concentration(14.8mmol/L).When the nitrogen concentration was above 74 mmol/L,the alga died.It indicated that high concentration of urea could inhibit the growth of Rholdella reticulata.
3.The nitrate reductase(NR)was rightly related to the biomass of the alga.Potassium nitrate had higher NR activity than urea had did.The enhanced activity of superoxide dismutase was due to an increase in concentration of nitrogen.
4.We used DEAE Sepharose Fast Flow ion exchange chromatography in combination of ammonium sulfate fractionation for purification of phycocyanin from Rhodella reticulata.The criterion of purity(R)was 3.92,the purified protein has two subunits with molecular mass of 18 and 22kDa each according to SDS-PAGE.The pure phycocyanin contained 17 kinds of amino acid.
5.The phycocyanin was more stable under unlighted,low temperature and neutrality pH.
6.For antioxidant activity assay on phycocyanin,the pure phycocyanin had higher antioxidant activity than crude phycocyanin had.Moreover,the pure phycocyanin exhibited effective antioxidant activity in a dose-dependent manner.
The unicellular red alga,Rhodella reticulata with a pyrenoid in it,can synthesize some bioactive substances such as exopolysaccharides,phycobiliprotein and pigments.In this paper,we investigated the optimization of nitrogen source,and made potassium nitrate and urea as a sole nitrogen source respectively,obtained the best nitrogen concentration,isolated and purified the phycocyanin from Rhodella reticulata,analyzed the amino acid composition and stability of the phycocyanin,as well as antioxidant activity of phycocyanin in vitro.The results were as follows:
1.Potassium nitrate as a nitrogen source,Rhodella reticulata grew well when nitrogen concentration was between 1.85～296mmol/L.The production of biomass and phycocyanin had increased 5.13%and 15.94%under the best nitrogen concentration(3.7mmol/L).When the nitrogen concentration was 296mmol/L,the maximum exopolysaccharides of Rhodella reticulata increased 254.87%.Therefore, potassium nitrate was the optimum nitrogen source.
2.Urea as a nitrogen source,Rhodella reticulata grew well when nitrogen concentration was between 1.85～29.6mmol/L,the production of biomass, exopolysaccharides and phycocyanin had increased 26.92%,23.58%and 35.22%at the best nitrogen concentration(14.8mmol/L).When the nitrogen concentration was above 74 mmol/L,the alga died.It indicated that high concentration of urea could inhibit the growth of Rholdella reticulata.
3.The nitrate reductase(NR)was rightly related to the biomass of the alga.Potassium nitrate had higher NR activity than urea had did.The enhanced activity of superoxide dismutase was due to an increase in concentration of nitrogen.
4.We used DEAE Sepharose Fast Flow ion exchange chromatography in combination of ammonium sulfate fractionation for purification of phycocyanin from Rhodella reticulata.The criterion of purity(R)was 3.92,the purified protein has two subunits with molecular mass of 18 and 22kDa each according to SDS-PAGE.The pure phycocyanin contained 17 kinds of amino acid.
5.The phycocyanin was more stable under unlighted,low temperature and neutrality pH.
6.For antioxidant activity assay on phycocyanin,the pure phycocyanin had higher antioxidant activity than crude phycocyanin had.Moreover,the pure phycocyanin exhibited effective antioxidant activity in a dose-dependent manner.
引文
[1]Viskari PJ,Colyer CL.Rapid extraction of phycobiliproteins from cultured cyanobacteria samples[J].Analytical Biochemistry.2003,319(2):263-271.
[2]Evans LV.Electron microscopical observations on a new red algal unicell,Rhodella maculata gen.nov.,sp.nov[J].European Journal of Phycology.1970,5(1):1-13.
[3]Deason TR,Butler GL,Rhyne C.Rhodella reticulata sp.nov.,a new coccoid rhodophytan alga(Porphyridiales)l,2[J].Journal of Phycology.1983,19(1):104-111.
[4]Yokoyama A,Sato K,Hara Y.The generic delimitation of Rhodella(Porphyridiales,Rhodophyta)with emphasis on ultrastructure and molecular phylogeny[J].Hydrobiologia.2004,512(1):177-183.
[5]Billard C,Fresnel J.Rhodella cyanea nov.sp.,une nouvelle Rhodophyceae unicellulaire[J].Cell Science.1986,302(7):271-276.
[6]Scott JL,Broadwater ST,Saunders BD,et al.Ultrastructure of vegetative organization and cell division in the unicellular red alga Dixoniella grisea gen.nov.(Rhodophyta)and a consideration of the genus Rhodella[J].Journal of phycology.1992,28(5):649-660.
[7]游文朗.蔷薇藻光生物反应器培养及其胞外多糖的研究[D].福建师范大学硕士学位论文.2006:19-20.
[8]Pekarkova B,Hindak F,Smarda J.Morphological characteristics and physiological properties of a coccoid rhodophycean alga Rhodella grisea from thermal springs at Piestany,Czechoslovakia[J].Archiv fur Protistenkunde.1988,135(1-4):69-83.
[9]Thomas JC,Passaquet C.Characterization of a phycoerythrin without α-subunits from a unicellular red alga[J].Journal of Biological Chemistry.1999,274(4):2472-2482.
[10]Kroen WK,Ramus J.Viscous polysaccharide and starch synthesis in Rhodella reticulata(Porphyridiales,Rhodophyta)l[J].Journal of Phycology.1990,26(2):266-271.
[11]Toncheva-Panova T,Ivanova J.A bacterial pathogen of Rhodella reticulata[J].Journal of Applied Microbiology.1997,83(6):707-711.
[12]Mihova SG,Georgiev DI,Minkova KM,et al.Phycobiliproteins in Rhodella reticulata and photoregulatory effects on their content[J].Journal of Biotechnology.1996,48(3):251-257.
[13] Arad S, Dubinsky 0, Simon B. A modified cell wall mutant of the red microalga Rhodella reticulata resistant to the herbicide 2, 6-dichlorobenzonitrile[J]. Journal of Phycology. 1993, 29(3):309-313.
[14] Geresh S, Arad S. The extracellular polysaccharides of the red microalgae: chemistry and rheology[J]. Bioresource technology. 1991, 38(2-3):195-201.
[15] Geresh S, Mamontov A, Weinstein J. Sulfation of extracellular polysaccharides of red microalgae: preparation, characterization and properties[J]. Journal of Biochemical and Biophysical Methods. 2002, 50(2-3): 179-187.
[16] Chow F, Oliveira MC, Pedersen M. In vitro assay and light regulation of nitrate reductase in red alga Gracilaria chilensis[J]. Journal of Plant Physiology. 2004,161(7):769-776.
[17] Edwards KF, Pfister CA, Van Alstyne KL. Nitrogen content in the brown alga Fucus gardneri and its relation to light, herbivory and wave exposure[J]. Journal of Experimental Marine Biology and Ecology. 2006, 336(1):99-109.
[18] Kakinuma M, Coury DA, Kuno Y, et al. Physiological and biochemical responses to thermal and salinity stresses in a sterile mutant of Ulva pertusa(Ulvales, Chlorophyta) [J]. Marine Biology. 2006,149(1):97-106.
[19] Wang R, Xing X, Crawford N. Nitrite acts as transcriptome signal at micromolar concentrations in Arabidopsis roots[J]. Plant Physiol. 2007.
[20] Pelah D, Marton I, Wang W, et al. Accumulation and protection activity of protease-resistant heat-stable proteins in Haematococcus pluvialis during high light and nitrogen starvation[J]. Journal of Applied Phycology. 2004,16(2):153-156.
[21] Orosa M, Torres E, Fidalgo P, et al. Production and analysis of secondary carotenoids in green algae[J]. Journal of Applied Phycology. 2000, 12(3):553-556.
[22] Huang JC, Chen F, Sandmann G. Stress-related differential expression of multiple β -carotene ketolase genes in the unicellular green alga Haematococcus pluvialis[J].Journal of Biotechnology. 2006, 122(2): 176-185.
[23] Khozin-Goldberg I, Shrestha P, Cohen Z. Mobilization of arachidonyl moieties from triacylglycerols into chloroplastic lipids following recovery from nitrogen starvation of the microalga Parietochloris incisa[J]. BBA-Molecular and Cell Biology of Lipids. 2005, 1738(1-3):63-71.
[24]Gomez PJL,Campo FE,Moreno DP,et al.Nitrogen availability influences the biochemical composition and photosynthesis of tank-cultivated Ulva rigida(Chlorophyta)[J].Journal of Applied Phycology.1998,10(4):383-389.
[25]Masojidek J,Torzillo G;Kopecky J,et al.Changes in chlorophyll fluorescence quenching and pigment composition in the green alga Chlorococcum sp.grown under nitrogen deficiency and salinity stress[J].Journal of Applied Phycology.2000,12(3):417-426.
[26]Sloth JK,Wiebe MG,Eriksen NT.Accumulation of phycocyanin in heterotrophic and mixotrophic cultures of the acidophilic red alga Galdieria sulphuraria[J].Enzyme and Microbial Technology.2006,38(1-2):168-175.[
27]Sanchez-Estudillo L,Freile-Pelegrin Y,Rivera-Madrid R,et al.Regulation of two photosynthetic pigment-related genes during stress-induced pigment formation in the green alga,Dunaliella salina[J].Biotechnology Letters.2006,28(11):787-791.
[28]邱昌恩,况琪军,刘国祥等.不同氮浓度对绿球藻生长及生理特性的影响[J].中国环境科学.2005,25(4):408-411.
[29]Lupi FM,Fernandes HML,Tom6 MM,et al Influence of nitrogen source and photoperiod on exopolysaccharide synthesis by the microalga Botryococcus braunii UC 58[J].Enzyme Microb.Technol.1994,16(7):546-550.
[30]张欣华,杨海波.不同培养条件对海洋微藻多糖含量的影响[J].生物学杂志.2000,17(6):17-18.
[31]张青田,董双林,胡桂坤等.不同氮源对微藻增殖的影响[J].海洋科学.2005,29(2):8-11.
[32]陈新美,梅兴国,房伟等.培养条件对螺旋藻生长和藻胆蛋白含量的影响[J].氨基酸和生物资源.2003,25(1):21-25.
[33]徐立,吴瑜端.有机氮化合物对海洋浮游植物生长的影响[J].厦门大学学报(自然科学版).1995,34(5):824-828.
[34]张清春,于仁诚,周名江等.不同氮源对微小亚历山大藻生长和毒素产生的影响[J].海洋学报.2005,27(6):138-145.
[35]胡晗华,石岩峻,丛威等.不同氮磷水平下中肋骨条藻对营养盐的吸收及光合 特性[J].应用与环境生物学报.2004,10(6):735-739。
[36]刘东艳,孙军,巩晶等.不同氮、磷比例对球等鞭金藻生长的影响[J].海洋水产研究.2002,23(1):29-32.
[37]李顺兴,郑凤英,洪华生等.氮磷营养盐与有机汞联合作用对微氏海链藻生长的影响[J].海洋科学.2005,29(10):36-39.
[38]Korbee N,Figueroa FL,Aguilera J.Effect of light quality on the accumulation of photosynthetic pigments,proteins and mycosporine-like amino acids in the red alga Porphyra leucosticta(Bangiales,Rhodophyta)[J].Journal of Photochemistry &Photobiology,B:Biology.2005,80(2):71-78.
[39]孙利芹,王长海,滕立.培养条件对紫球藻B.藻红蛋白含量的影响[J].浙江大学学报(农业与生命科学版).2007,33(1):40-44.
[40]张曼,曾波,王明书等.温度升高对高光强环境下蛋白核小球藻(Chlolorella pyrenoidosa)光能利用和生长的阻抑效应[J].生态学报.2007,27(2):662-667.
[41]张少斌,燕安,刘慧等.螺旋藻多糖和藻胆蛋白对人白血病Jurkat细胞生长的影响[J].中国现代医学杂志.2006,16(14):2115-2117,2120.
[42]于娟,唐学玺,张培玉等.CO_2加富对两种海洋微绿藻的生长、光合作用和抗氧化酶活性的影响[J].生态学报.2005,25(2):197-202.
[43]刘春颖,张正斌,陈小睿.一氧化氮和铁对海洋微藻生长的交互影响[J].海洋学报.2005,27(6):123-130.
[44]邱昌恩,刘国祥,况琪军等.Cu~(2+)对一种绿球藻生长及生理特性的影响[J].应用与环境生物学报.2005,11(6):690-693.
[45]刘洁,常学秀,黄丽娟等.Ni元素对铜绿微囊藻的生长、光谱特性及藻胆蛋白含量的影响『J].云南大学学报(自然科学版).2005,27(4):365-368.
[46]Kakita H,Kamishima H.Effects of environmental factors and metal ions on growth of the red alga Gracilaria Chorda Holmes(Gracilariales,Rhodophyta)[J].Journal of Applied Phycology.2006,18(3):469-474.
[47]周志刚,尹长松。富硒极大螺旋藻整细胞、藻胆体及藻蓝蛋白的光谱特性[J].上海水产大学学报.2002,11(3):208-214.
[48]岳霞丽,张新萍,胡先文等.苄嘧磺隆对蛋白核小球藻的生长效应研究[J].中国农业科学.2006,39(9):1823-1827.
[49]Rajendran UM,Kathirvel E,Narayanaswamy A.Effects of a fungicide,an insecticide,and a biopesticide on Tolypothrix scytonemoides[J].Pesticide Biochemistry and Physiology.2007,87(2):164-171.
[50]郑必胜,郭祀远,尤珊.磁场处理对螺旋藻生长及胞外多糖的影响[J].食品科学.2007,28(1):94-98.
[51]王培磊,张学成,孟振等.乙酸对两种杜氏藻生长和细胞生化组成的影响[J].海洋科学。2005,29(6):22-27.
[52]Corzo A,Niell FX.Nitrate-reductase activity and in vivo nitrate-reduction rate in Ulva rigida illuminated by blue light[J].Marine Biology.1994,120(1):17-23.
[53]Fatima LP,Cabral OM,Colepicolo P.Characterization and daily variation of nitrate reductase in Gracilaria tenuistipitata(Rhodophyta)[J].Biochemical and Biophysical Research Communications.2002,295(1):50-54.
[54]Sun Y,Gao X,Li Q,et al.Functional complementation of a nitrate reductase defective mutant of a green alga Dunaliella viridis by introducing the nitrate reductase gene[J].Gene.2006,377:140-149.
[55]Flores E,Frias JE,Rubio LM,et al.Photosynthetic nitrate assimilation in cyanobacteria[J].Photosynthesis Research.2005,83(2):117-133.
[56]Han TKSH,Park JS,Lee HK,et al.Physiological responses of Ulva pertusa and U.armoricana to copper exposure[J].Aquatic Toxicology.2007,66:101-108.
[57]Touchette BW.Seagrass-salinity interactions:Physiological mechanisms used by submersed marine angiosperms for a life at sea[J].Journal of Experimental Marine Biology and Ecology.2007,350(1-2):194-215.
[58]Pokora W,Ji IR,Tuka Z.Activities of superoxide dismutase(SOD)isoforms during growth of Scenedesmus(Chlorophyta)species and strains grown in batch-cultures[J].Acta physiologiae plantarum.2003,25(4):375-384.
[59]Augusti PR,Conterato GM,Somacal S,et al.Effect of astaxanthin on kidney function impairment and oxidative stress induced by mercuric chloride in rats[J].Food Chem Toxicol.2007.
[60]Wang G,Hu C,Li D,et al.The response of antioxidant systems in Nostoc sphaeroides against UV-B radiation and the protective effects of exogenous antioxidants[J].Advances in Space Research.2007,39(6):1034-1042.
[61]Hong Y,Xie X,Li FM.Responses of enzymatic antioxidants and non-enzymatic antioxidants in the cyanobacterium Microcystis aeruginosa to the allelochemical ethyl 2-methyl acetoacetate(EMA)isolated from reed(Phragmites communis)[J].Journal of Plant Physiology.2007,9:218-227.
[62]Rossa MM,Oliveira MC,Okamoto OK,et al.Effect of visible light on superoxide dismutase(SOD)activity in the red alga Gracilariopsis tenuifrons(Gracilariales,Rhodophyta)[J].Journal of Applied Phycology.2002,14(3):151-157.
[63]Guzman-Murillo MA,1.6pez-Bolanos CC,Ledesma-Verdejo T,et al.Effects of fertilizer-based culture media on the production of exocellular polysaccharides and cellular superoxide dismutase by Phaeodactylurn tricornuturn(Bohlin)[J].Journal of Applied Phycology.2007,19(1):33-41.
[64]Tang D,Shi S,Li D,et al.Physiological and biochemical responses of Scytonema javanicum(cyanobacterium)to salt stress[J].Journal of Arid Environments.2007,71(3):312-320.
[65]Zbigniew T,Wojciech P.Individual and combined effect of anthracene,cadmium,and chloridazone on growth and activity of SOD izoformes in three Scenedesmus species[J].Ecotoxicology and Environmental Safety.2006,65(3):323-331.
[66]MacColl R.Cyanobacterial phycobilisomes[J].Journal of Structural Biology.1998,124(2-3):311-334.
[67]Grossman AR,Schaefer MR,Chiang GG,et al.The phycobilisome,a light-harvesting complex responsive to environmental conditions[J].Microbiol Rev.1993,57(3):725-749.
[68]王广策,邓田.藻胆蛋白的研究概况(Ⅰ):--藻胆蛋白的种类与组成[J].海洋科学.2000,24(2):22-25.
[69]王广策,邓田.藻胆蛋白的研究概况(Ⅱ)-藻胆蛋白的结构及其光谱特性….海洋科学.2000,24(3):19-22.
[70]马圣媛,王广策,孙海宝等.藻胆蛋白的研究概况Ⅲ.蓝藻藻胆蛋白合成的分子机制及调控[J].海洋科学.2002,26(8):40-43.
[71]Arciero DM,Dallas JL,Glazer AN.In vitro attachment of bilins to apophycocyanin. Ⅲ.Properties of the phycoerythrobilin adduct[J].Journal of Biological Chemistry.1988,263(34):18358-18363.
[72]Benedetti S,Rinalducci S,Benvenuti F,et al.Purification and characterization of phycocyanin from the blue-green alga Aphanizomenon flos-aquae[J].Journal of Chromatography B.2006,833(1):12-18.
[73]Glazer AN.Light guides.Directional energy transfer in a photosynthetic antenna[J].Journal of Biological Chemistry.1989,264(1):1-4.
[74]Niu JF,Wang GC,Tseng CK.Method for large-scale isolation and purification of R-phycoerythrin from red alga Polysiphonia urceolata Grev[J].Protein Expression and Purification.2006,49(1):23-31.
[75]Patil G,Chethana S,Sridevi AS,et al.Method to obtain C-phycocyanin of high purity[J].Journal of Chromatography A.2006,1127(1-2):76-81.
[76]Madhyastha HK,Radha KS,Sugiki M,et al.Purification of c-phycocyanin from Spirulina fusiformis and its effect on the induction of urokinase-type plasminogen activator from calf pulmonary endothelial cells[J].Phytomedicine.2006,13(8):564-569.
[77]Soni B,Kalavadia B,Trivedi U,et al.Extraction,purification and characterization of phycocyanin from Oscillatoria quadripunctulata-Isolated from the rocky shores of Bet-Dwarka,Gujarat,India[J].Process Biochemistry.2006,41(9):2017-2023.
[78]Soni B,Trivedi U,Madamwar D.A novel method of single step hydrophobic interaction chromatography for the purification of phycocyanin from Phormidium fragile and its characterization for antioxidant property[J].Bioresource Technology.2008,99(1):188-194.
[79]Rito-Palomares M,Nunez L,Amador D.Practical application of aqueous two-phase systems for the development of a prototype process for c-phycocyanin recovery from Spirulina maxima[J].Journal of Chemical Technology and Biotechnology.2001,76(12):1273-1280.
[80]王宇飞,周明,赵开弘等.层理鞭枝藻藻红蓝蛋白α亚基的固定化与光化学稳定性研究[J].功能高分子学报.2004,17(3):411-416.
[81]彭卫民,商树田.钝顶螺旋藻别藻蓝蛋白的稳定性研究[J].中国水产.1999,(7):24-25.
[82]吕彩真,欧光南.红毛菜藻红蛋白的提取及稳定性研究[J].集美大学学报(自然科学版).2002,7(1):15-19.
[83]张少斌,张英娇,刘慧等。螺旋藻别藻蓝蛋白稳定性研究[J].食品研究与开发。2007,28(2):21-23.
[84]Furuki T,Maeda S,Imajo S,et al.Rapid and selective extraction of phycocyanin from Spirulina platensis with ultrasonic cell disruption[J].Journal of Applied Phycology.2003,15(4):319-324.
[85]王仲孚,田庚元.藻胆蛋白研究[J].生命的化学.2000,20(2):72-75.
[86]余九九,李宽钰.螺旋藻的藻胆蛋白提取及稳定性研究[J].海洋通报.1997,16(4):26-28.
[87]Jongjareonrak A,Benjakul S,Visessanguan W,et al.Antioxidative activity and properties of fish skin gelatin films incorporated with BHT and a-tocopherol[J].Food Hydrocolloids.2008,22(3):449-458.
[88]Lee HW,Chien JT,Chen BH.Inhibition of cholesterol oxidation in marinated foods as affected by antioxidants during heating[J].Food Chemistry.2008,108(1):234-244.
[89]Passone MA,Resnik S,Etcheverry MG.Antiaflatoxigenic property of food grade antioxidants under different conditions of water activity in peanut grains[J].Int J Food Microbiol.2007,118(1):8-14.
[90]Apel K,Hirt H.Reactive oxygen species:metabolism,oxidative stress,and signal transduction[J].Annual Review of Plant Biology.2004,55(1):373-399.
[91]Jose N,Janardhanan KK.Antioxidant and antitumour activity of Pleurotus florida[J].Current Science.2000,79(7):941-943.
[92]Morcos NC,Berns M,Henry WL.Phycocyanin:laser activation,cytotoxic effect,and uptake in human atherosclerotic plaque[J].Lasers Surg Med.1988,8(1):7-10.
[93]Galli F,Canestrari F,Bellomo G.Pathophysiology of the oxidative stress and its implication in uremia and dialysis[J].Contrib Nephrol.1999,127:1-31.
[94]Ghiselli A,Serafini M,Maiani G,et al.A fluorescence-based method for measuring total plasma antioxidant capability[J].Free Radical Biology and Medicine.1995,18(1):29-36.
[95]Benedetti S,Benvenuti F,Pagliarani S,et al.Antioxidant properties of a novel phycocyanin extract from the blue-green alga Aphanizomenon flos-aquae[J].Life Sciences.2004,75(19):2353-2362.
[96]Pinero EJE,Bermejo BP,Villar FAM.Antioxidant activity of different fractions of Spirulina platensis protean extract[J].I1 Farmaco.2001,56(5-7):497-500.
[97]魏文志,夏文水,吴玉娟.小球藻蛋白质的分离、纯化及抗氧化特性[J].上海水产大学学报.2007,16(2):135-139。
[98]杨立红,王晓洁,钟旭升等.鱼腥藻藻蓝蛋白的抗氧化作用[J].食品科学.2006,27(12):208-213.
[99]唐玫,王曼,郭宝江.富硒藻蓝蛋白对小鼠免疫功能及抗氧化活性的影响[J].营养学报.2001,23(3):275-278.
[100]陈美珍,张永雨,余杰等.龙须菜藻胆蛋白的分离及其清除自由基作用的初步研究[J].食品科学.2004,25(3):159-162.
[101]陈美珍,余杰,钟秋玲等。龙须菜藻胆蛋白免疫功能和抗氧化作用的研究[J].食品科学.2005,26(9):456.459.
[102]Romay C,GonzAlez R.Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals[J].Journal of Pharmacy and Pharmacology.2000,52(4):367-368.
[103]Farooq SM,Asokan D,Sakthivel R,et al.Salubrious effect of C-phycocyanin against oxalate-mediated renal cell injury[J].Clinica Chimica Acta.2004,348(1-2):199-205.
[104]Bhat VB,Madyastha KM.C-phycocyanin:a potent peroxyl radical scavenger in vivo and in vitro[J].Biochemical and Biophysical Research Communications.2000,275(1):20-25.
[105]Romay C,Armesto J,Remirez D,et al.Antioxidant and anti-inflammatory properties of C-phycocyanin from blue-green algae[J].Inflammation Research.1998,47(1):36-41.
[106]Romay C,Gonzalez R,Ledon N,et al.C-phycocyanin:a biliprotein with antioxidant,anti-inflammatory and neuroprotective effects[J].Curr Protein Pept Sci.2003,4(3):207-216.
[107]Cherng SC,Cheng SN,Tam A,et al.Anti-inflammatory activity of c-phycocyanin in lipopolysaccharide-stimulated RAW 264.7 macrophages[J].Life Sci.2007,81(19-20):1431-1435.
[108]何立明,郑树.藻蓝蛋白对大肠癌和S180小鼠移植瘤的光敏作用[J].激光医学.1996,6(4):156-157,160.
[109]张成武,曾昭琪.藻蓝蛋白对小鼠粒单系祖细胞生成的影响[J]。中国海洋药物.1996,15(4):25-28.
[110]关燕清,周天鸿.光固定化藻蓝蛋白对内皮细胞生长的影响[J]。功能高分子学报.2003,16(3):373-376.
[111]关燕清,徐和德.光固定化藻蓝蛋白对体外肝癌细胞7402的抑制作用[J].离子交换与吸附.2000,16(6):547-552.
[112]张永雨,陈美珍,余杰等.龙须菜藻胆蛋白抗突变与抗肿瘤作用的研究[J].中国海洋药物.2005,24(3):36-38.
[113]张惟杰.糖复合物生化研究技术(第二版)[M].杭州:浙江大学出版社,1999.11-17
[114]Tandeau MN,Houmard J.Complementary chromatic adaptation:physiological conditions and action spectra[J].Methods Enzymol.1988,167:318-328.
[115]张志良.植物生理学实验指导[M].北京:高等教育出版社,1990。
[116]Chang FH,McClean M.Growth responses ofAlexandrium minutum(Dinophyceae)as a function of three different nitrogen sources and irradiance[J].New Zealand Journal of Marine and Freshwater Research.1997,31(1):1-7.
[117]赵素芬,孙会强.氮源、磷源及重金属离子对绿色巴夫藻生长的影响[J].湛江海洋大学学报.2005,25(1):60-63.
[118]Turpin DH.Effects of inorganic N availability on algal photosynthesis and carbon metabolism[J].Journal of Phycology.1991,27(1):14-20.
[119]Rayburn WR,Kroen WK.Influence of growth status and nutrients on extracellular polysaccharide synthesis by the soil alga Chlamydornonas mexicana(Chlorophyceae)[J].Journal of Phycology.1984,20(2):253-257.
[120]Sloth JK,Eriksen NT.Accumulation of phycocyanin in heterotrophic and mixotrophic cultures of the acidophilic red alga Galdieria sulphuraria[J].Enzyme and Microbial Technology.2006,38(1-2):168-175.
[121]孙辉,徐文华,雷高鹏等.氮、磷、硫对盐生杜氏藻色素积累的影响[J].四川大学学报(自然科学版).2005,42(2):403-407.
[122]蔡恒江,唐学玺,张培玉等.UV-B辐射对青岛大扁藻生长及其某些生理特性的影响[J]。海洋科学进展.2005,23(4):460-465.
[123]Dortch Q,Ahmed SI,Packard TT.Nitrate reductase and glutamate dehydrogenase activities in Skeletonema costatum as measures of nitrogen assimilation rates 1[J].Journal of Plankton Research.2002,1(2):169-186.
[124]李梅燕.西施舌人工育苗饵料藻种的选育[D].福建师范大学硕士学位论文2004:64-65.
[125]Lomas MW.Nitrate reductase and urease enzyme activity in the marine diatom Thalassiosira weissflogii(Bacillariophyceae):interactions among nitrogen substrates[J].Marine Biology.2004,144(1):37-44.
[126]Misra N,Gupta AK.Effect of salinity and different nitrogen sources on the activity of antioxidant enzymes and indole alkaloid content in Catharanthus roseus seedlings[J].Journal of Plant Physiology.2006,163(1):11-18.
[127]章申峰.C-藻蓝蛋白及其亚基的分离纯化和体外抗肿瘤研究[D].浙江大学硕士学位论文.2005:39-41.
[128]Minkova KM,Tchorbadjieva MI,Fournadjieva ST,et al.Purification of C-phycocyanin from Spirulina(Arthrospira)fusiformis[J].Journal of Biotechnology.2003,102:55-59.
[129]欧瑜,董剑,吴梧桐.盐生隐杆藻藻胆蛋白的分离纯化和特性研究[J].药物生物技术.2004,11(1):37-41.
[130]殷钢,李琛.螺旋藻中藻胆蛋白和多糖的分离纯化及产品特性[J].精细化工.1999,16(2):10-13.
[131]Sarada RMGP,Ravishankar GA.Phycocyanin from Spirulina sp influnence of processing of biomass on phycocyanin yield analysis of efficacy of extraction methods and stability studies on phycocyanin[J].Process Biochemistry.1999,34(8):795-801.
[132]范刚,陈德文,潘思铁等.葛仙米藻胆蛋白提取工艺及藻蓝蛋白稳定性研究[J].食品科学.2005,26(9):215-218.
[133]杨芬,张瑞萍,贺玖明等.羟自由基的产生、捕集及检测方法[J].药学学报. 2007,42(7):692-697.
[134]Huycke MM,Abrams Victoria,Moore Danny Ro Enterococcus faecalis produces extracellular superoxide and hydrogen peroxide that damages colonic epithelial cell[J].DNA Carcinogenesis.2002,23(3):529-536.
[135]Wolff SP.Ferrous ion oxidation in presence of ferric ion indicator xylenol orange for measurement of hydroperoxides[J].Mothods in enzymology.1994,233(18):183-189.
[136]周站平,陈秀兰,陈超等.藻胆蛋白脱辅基蛋白对其抗氧化活性的影响[J].海洋科学.2003,27(5):77-80.
[2]Evans LV.Electron microscopical observations on a new red algal unicell,Rhodella maculata gen.nov.,sp.nov[J].European Journal of Phycology.1970,5(1):1-13.
[3]Deason TR,Butler GL,Rhyne C.Rhodella reticulata sp.nov.,a new coccoid rhodophytan alga(Porphyridiales)l,2[J].Journal of Phycology.1983,19(1):104-111.
[4]Yokoyama A,Sato K,Hara Y.The generic delimitation of Rhodella(Porphyridiales,Rhodophyta)with emphasis on ultrastructure and molecular phylogeny[J].Hydrobiologia.2004,512(1):177-183.
[5]Billard C,Fresnel J.Rhodella cyanea nov.sp.,une nouvelle Rhodophyceae unicellulaire[J].Cell Science.1986,302(7):271-276.
[6]Scott JL,Broadwater ST,Saunders BD,et al.Ultrastructure of vegetative organization and cell division in the unicellular red alga Dixoniella grisea gen.nov.(Rhodophyta)and a consideration of the genus Rhodella[J].Journal of phycology.1992,28(5):649-660.
[7]游文朗.蔷薇藻光生物反应器培养及其胞外多糖的研究[D].福建师范大学硕士学位论文.2006:19-20.
[8]Pekarkova B,Hindak F,Smarda J.Morphological characteristics and physiological properties of a coccoid rhodophycean alga Rhodella grisea from thermal springs at Piestany,Czechoslovakia[J].Archiv fur Protistenkunde.1988,135(1-4):69-83.
[9]Thomas JC,Passaquet C.Characterization of a phycoerythrin without α-subunits from a unicellular red alga[J].Journal of Biological Chemistry.1999,274(4):2472-2482.
[10]Kroen WK,Ramus J.Viscous polysaccharide and starch synthesis in Rhodella reticulata(Porphyridiales,Rhodophyta)l[J].Journal of Phycology.1990,26(2):266-271.
[11]Toncheva-Panova T,Ivanova J.A bacterial pathogen of Rhodella reticulata[J].Journal of Applied Microbiology.1997,83(6):707-711.
[12]Mihova SG,Georgiev DI,Minkova KM,et al.Phycobiliproteins in Rhodella reticulata and photoregulatory effects on their content[J].Journal of Biotechnology.1996,48(3):251-257.
[13] Arad S, Dubinsky 0, Simon B. A modified cell wall mutant of the red microalga Rhodella reticulata resistant to the herbicide 2, 6-dichlorobenzonitrile[J]. Journal of Phycology. 1993, 29(3):309-313.
[14] Geresh S, Arad S. The extracellular polysaccharides of the red microalgae: chemistry and rheology[J]. Bioresource technology. 1991, 38(2-3):195-201.
[15] Geresh S, Mamontov A, Weinstein J. Sulfation of extracellular polysaccharides of red microalgae: preparation, characterization and properties[J]. Journal of Biochemical and Biophysical Methods. 2002, 50(2-3): 179-187.
[16] Chow F, Oliveira MC, Pedersen M. In vitro assay and light regulation of nitrate reductase in red alga Gracilaria chilensis[J]. Journal of Plant Physiology. 2004,161(7):769-776.
[17] Edwards KF, Pfister CA, Van Alstyne KL. Nitrogen content in the brown alga Fucus gardneri and its relation to light, herbivory and wave exposure[J]. Journal of Experimental Marine Biology and Ecology. 2006, 336(1):99-109.
[18] Kakinuma M, Coury DA, Kuno Y, et al. Physiological and biochemical responses to thermal and salinity stresses in a sterile mutant of Ulva pertusa(Ulvales, Chlorophyta) [J]. Marine Biology. 2006,149(1):97-106.
[19] Wang R, Xing X, Crawford N. Nitrite acts as transcriptome signal at micromolar concentrations in Arabidopsis roots[J]. Plant Physiol. 2007.
[20] Pelah D, Marton I, Wang W, et al. Accumulation and protection activity of protease-resistant heat-stable proteins in Haematococcus pluvialis during high light and nitrogen starvation[J]. Journal of Applied Phycology. 2004,16(2):153-156.
[21] Orosa M, Torres E, Fidalgo P, et al. Production and analysis of secondary carotenoids in green algae[J]. Journal of Applied Phycology. 2000, 12(3):553-556.
[22] Huang JC, Chen F, Sandmann G. Stress-related differential expression of multiple β -carotene ketolase genes in the unicellular green alga Haematococcus pluvialis[J].Journal of Biotechnology. 2006, 122(2): 176-185.
[23] Khozin-Goldberg I, Shrestha P, Cohen Z. Mobilization of arachidonyl moieties from triacylglycerols into chloroplastic lipids following recovery from nitrogen starvation of the microalga Parietochloris incisa[J]. BBA-Molecular and Cell Biology of Lipids. 2005, 1738(1-3):63-71.
[24]Gomez PJL,Campo FE,Moreno DP,et al.Nitrogen availability influences the biochemical composition and photosynthesis of tank-cultivated Ulva rigida(Chlorophyta)[J].Journal of Applied Phycology.1998,10(4):383-389.
[25]Masojidek J,Torzillo G;Kopecky J,et al.Changes in chlorophyll fluorescence quenching and pigment composition in the green alga Chlorococcum sp.grown under nitrogen deficiency and salinity stress[J].Journal of Applied Phycology.2000,12(3):417-426.
[26]Sloth JK,Wiebe MG,Eriksen NT.Accumulation of phycocyanin in heterotrophic and mixotrophic cultures of the acidophilic red alga Galdieria sulphuraria[J].Enzyme and Microbial Technology.2006,38(1-2):168-175.[
27]Sanchez-Estudillo L,Freile-Pelegrin Y,Rivera-Madrid R,et al.Regulation of two photosynthetic pigment-related genes during stress-induced pigment formation in the green alga,Dunaliella salina[J].Biotechnology Letters.2006,28(11):787-791.
[28]邱昌恩,况琪军,刘国祥等.不同氮浓度对绿球藻生长及生理特性的影响[J].中国环境科学.2005,25(4):408-411.
[29]Lupi FM,Fernandes HML,Tom6 MM,et al Influence of nitrogen source and photoperiod on exopolysaccharide synthesis by the microalga Botryococcus braunii UC 58[J].Enzyme Microb.Technol.1994,16(7):546-550.
[30]张欣华,杨海波.不同培养条件对海洋微藻多糖含量的影响[J].生物学杂志.2000,17(6):17-18.
[31]张青田,董双林,胡桂坤等.不同氮源对微藻增殖的影响[J].海洋科学.2005,29(2):8-11.
[32]陈新美,梅兴国,房伟等.培养条件对螺旋藻生长和藻胆蛋白含量的影响[J].氨基酸和生物资源.2003,25(1):21-25.
[33]徐立,吴瑜端.有机氮化合物对海洋浮游植物生长的影响[J].厦门大学学报(自然科学版).1995,34(5):824-828.
[34]张清春,于仁诚,周名江等.不同氮源对微小亚历山大藻生长和毒素产生的影响[J].海洋学报.2005,27(6):138-145.
[35]胡晗华,石岩峻,丛威等.不同氮磷水平下中肋骨条藻对营养盐的吸收及光合 特性[J].应用与环境生物学报.2004,10(6):735-739。
[36]刘东艳,孙军,巩晶等.不同氮、磷比例对球等鞭金藻生长的影响[J].海洋水产研究.2002,23(1):29-32.
[37]李顺兴,郑凤英,洪华生等.氮磷营养盐与有机汞联合作用对微氏海链藻生长的影响[J].海洋科学.2005,29(10):36-39.
[38]Korbee N,Figueroa FL,Aguilera J.Effect of light quality on the accumulation of photosynthetic pigments,proteins and mycosporine-like amino acids in the red alga Porphyra leucosticta(Bangiales,Rhodophyta)[J].Journal of Photochemistry &Photobiology,B:Biology.2005,80(2):71-78.
[39]孙利芹,王长海,滕立.培养条件对紫球藻B.藻红蛋白含量的影响[J].浙江大学学报(农业与生命科学版).2007,33(1):40-44.
[40]张曼,曾波,王明书等.温度升高对高光强环境下蛋白核小球藻(Chlolorella pyrenoidosa)光能利用和生长的阻抑效应[J].生态学报.2007,27(2):662-667.
[41]张少斌,燕安,刘慧等.螺旋藻多糖和藻胆蛋白对人白血病Jurkat细胞生长的影响[J].中国现代医学杂志.2006,16(14):2115-2117,2120.
[42]于娟,唐学玺,张培玉等.CO_2加富对两种海洋微绿藻的生长、光合作用和抗氧化酶活性的影响[J].生态学报.2005,25(2):197-202.
[43]刘春颖,张正斌,陈小睿.一氧化氮和铁对海洋微藻生长的交互影响[J].海洋学报.2005,27(6):123-130.
[44]邱昌恩,刘国祥,况琪军等.Cu~(2+)对一种绿球藻生长及生理特性的影响[J].应用与环境生物学报.2005,11(6):690-693.
[45]刘洁,常学秀,黄丽娟等.Ni元素对铜绿微囊藻的生长、光谱特性及藻胆蛋白含量的影响『J].云南大学学报(自然科学版).2005,27(4):365-368.
[46]Kakita H,Kamishima H.Effects of environmental factors and metal ions on growth of the red alga Gracilaria Chorda Holmes(Gracilariales,Rhodophyta)[J].Journal of Applied Phycology.2006,18(3):469-474.
[47]周志刚,尹长松。富硒极大螺旋藻整细胞、藻胆体及藻蓝蛋白的光谱特性[J].上海水产大学学报.2002,11(3):208-214.
[48]岳霞丽,张新萍,胡先文等.苄嘧磺隆对蛋白核小球藻的生长效应研究[J].中国农业科学.2006,39(9):1823-1827.
[49]Rajendran UM,Kathirvel E,Narayanaswamy A.Effects of a fungicide,an insecticide,and a biopesticide on Tolypothrix scytonemoides[J].Pesticide Biochemistry and Physiology.2007,87(2):164-171.
[50]郑必胜,郭祀远,尤珊.磁场处理对螺旋藻生长及胞外多糖的影响[J].食品科学.2007,28(1):94-98.
[51]王培磊,张学成,孟振等.乙酸对两种杜氏藻生长和细胞生化组成的影响[J].海洋科学。2005,29(6):22-27.
[52]Corzo A,Niell FX.Nitrate-reductase activity and in vivo nitrate-reduction rate in Ulva rigida illuminated by blue light[J].Marine Biology.1994,120(1):17-23.
[53]Fatima LP,Cabral OM,Colepicolo P.Characterization and daily variation of nitrate reductase in Gracilaria tenuistipitata(Rhodophyta)[J].Biochemical and Biophysical Research Communications.2002,295(1):50-54.
[54]Sun Y,Gao X,Li Q,et al.Functional complementation of a nitrate reductase defective mutant of a green alga Dunaliella viridis by introducing the nitrate reductase gene[J].Gene.2006,377:140-149.
[55]Flores E,Frias JE,Rubio LM,et al.Photosynthetic nitrate assimilation in cyanobacteria[J].Photosynthesis Research.2005,83(2):117-133.
[56]Han TKSH,Park JS,Lee HK,et al.Physiological responses of Ulva pertusa and U.armoricana to copper exposure[J].Aquatic Toxicology.2007,66:101-108.
[57]Touchette BW.Seagrass-salinity interactions:Physiological mechanisms used by submersed marine angiosperms for a life at sea[J].Journal of Experimental Marine Biology and Ecology.2007,350(1-2):194-215.
[58]Pokora W,Ji IR,Tuka Z.Activities of superoxide dismutase(SOD)isoforms during growth of Scenedesmus(Chlorophyta)species and strains grown in batch-cultures[J].Acta physiologiae plantarum.2003,25(4):375-384.
[59]Augusti PR,Conterato GM,Somacal S,et al.Effect of astaxanthin on kidney function impairment and oxidative stress induced by mercuric chloride in rats[J].Food Chem Toxicol.2007.
[60]Wang G,Hu C,Li D,et al.The response of antioxidant systems in Nostoc sphaeroides against UV-B radiation and the protective effects of exogenous antioxidants[J].Advances in Space Research.2007,39(6):1034-1042.
[61]Hong Y,Xie X,Li FM.Responses of enzymatic antioxidants and non-enzymatic antioxidants in the cyanobacterium Microcystis aeruginosa to the allelochemical ethyl 2-methyl acetoacetate(EMA)isolated from reed(Phragmites communis)[J].Journal of Plant Physiology.2007,9:218-227.
[62]Rossa MM,Oliveira MC,Okamoto OK,et al.Effect of visible light on superoxide dismutase(SOD)activity in the red alga Gracilariopsis tenuifrons(Gracilariales,Rhodophyta)[J].Journal of Applied Phycology.2002,14(3):151-157.
[63]Guzman-Murillo MA,1.6pez-Bolanos CC,Ledesma-Verdejo T,et al.Effects of fertilizer-based culture media on the production of exocellular polysaccharides and cellular superoxide dismutase by Phaeodactylurn tricornuturn(Bohlin)[J].Journal of Applied Phycology.2007,19(1):33-41.
[64]Tang D,Shi S,Li D,et al.Physiological and biochemical responses of Scytonema javanicum(cyanobacterium)to salt stress[J].Journal of Arid Environments.2007,71(3):312-320.
[65]Zbigniew T,Wojciech P.Individual and combined effect of anthracene,cadmium,and chloridazone on growth and activity of SOD izoformes in three Scenedesmus species[J].Ecotoxicology and Environmental Safety.2006,65(3):323-331.
[66]MacColl R.Cyanobacterial phycobilisomes[J].Journal of Structural Biology.1998,124(2-3):311-334.
[67]Grossman AR,Schaefer MR,Chiang GG,et al.The phycobilisome,a light-harvesting complex responsive to environmental conditions[J].Microbiol Rev.1993,57(3):725-749.
[68]王广策,邓田.藻胆蛋白的研究概况(Ⅰ):--藻胆蛋白的种类与组成[J].海洋科学.2000,24(2):22-25.
[69]王广策,邓田.藻胆蛋白的研究概况(Ⅱ)-藻胆蛋白的结构及其光谱特性….海洋科学.2000,24(3):19-22.
[70]马圣媛,王广策,孙海宝等.藻胆蛋白的研究概况Ⅲ.蓝藻藻胆蛋白合成的分子机制及调控[J].海洋科学.2002,26(8):40-43.
[71]Arciero DM,Dallas JL,Glazer AN.In vitro attachment of bilins to apophycocyanin. Ⅲ.Properties of the phycoerythrobilin adduct[J].Journal of Biological Chemistry.1988,263(34):18358-18363.
[72]Benedetti S,Rinalducci S,Benvenuti F,et al.Purification and characterization of phycocyanin from the blue-green alga Aphanizomenon flos-aquae[J].Journal of Chromatography B.2006,833(1):12-18.
[73]Glazer AN.Light guides.Directional energy transfer in a photosynthetic antenna[J].Journal of Biological Chemistry.1989,264(1):1-4.
[74]Niu JF,Wang GC,Tseng CK.Method for large-scale isolation and purification of R-phycoerythrin from red alga Polysiphonia urceolata Grev[J].Protein Expression and Purification.2006,49(1):23-31.
[75]Patil G,Chethana S,Sridevi AS,et al.Method to obtain C-phycocyanin of high purity[J].Journal of Chromatography A.2006,1127(1-2):76-81.
[76]Madhyastha HK,Radha KS,Sugiki M,et al.Purification of c-phycocyanin from Spirulina fusiformis and its effect on the induction of urokinase-type plasminogen activator from calf pulmonary endothelial cells[J].Phytomedicine.2006,13(8):564-569.
[77]Soni B,Kalavadia B,Trivedi U,et al.Extraction,purification and characterization of phycocyanin from Oscillatoria quadripunctulata-Isolated from the rocky shores of Bet-Dwarka,Gujarat,India[J].Process Biochemistry.2006,41(9):2017-2023.
[78]Soni B,Trivedi U,Madamwar D.A novel method of single step hydrophobic interaction chromatography for the purification of phycocyanin from Phormidium fragile and its characterization for antioxidant property[J].Bioresource Technology.2008,99(1):188-194.
[79]Rito-Palomares M,Nunez L,Amador D.Practical application of aqueous two-phase systems for the development of a prototype process for c-phycocyanin recovery from Spirulina maxima[J].Journal of Chemical Technology and Biotechnology.2001,76(12):1273-1280.
[80]王宇飞,周明,赵开弘等.层理鞭枝藻藻红蓝蛋白α亚基的固定化与光化学稳定性研究[J].功能高分子学报.2004,17(3):411-416.
[81]彭卫民,商树田.钝顶螺旋藻别藻蓝蛋白的稳定性研究[J].中国水产.1999,(7):24-25.
[82]吕彩真,欧光南.红毛菜藻红蛋白的提取及稳定性研究[J].集美大学学报(自然科学版).2002,7(1):15-19.
[83]张少斌,张英娇,刘慧等。螺旋藻别藻蓝蛋白稳定性研究[J].食品研究与开发。2007,28(2):21-23.
[84]Furuki T,Maeda S,Imajo S,et al.Rapid and selective extraction of phycocyanin from Spirulina platensis with ultrasonic cell disruption[J].Journal of Applied Phycology.2003,15(4):319-324.
[85]王仲孚,田庚元.藻胆蛋白研究[J].生命的化学.2000,20(2):72-75.
[86]余九九,李宽钰.螺旋藻的藻胆蛋白提取及稳定性研究[J].海洋通报.1997,16(4):26-28.
[87]Jongjareonrak A,Benjakul S,Visessanguan W,et al.Antioxidative activity and properties of fish skin gelatin films incorporated with BHT and a-tocopherol[J].Food Hydrocolloids.2008,22(3):449-458.
[88]Lee HW,Chien JT,Chen BH.Inhibition of cholesterol oxidation in marinated foods as affected by antioxidants during heating[J].Food Chemistry.2008,108(1):234-244.
[89]Passone MA,Resnik S,Etcheverry MG.Antiaflatoxigenic property of food grade antioxidants under different conditions of water activity in peanut grains[J].Int J Food Microbiol.2007,118(1):8-14.
[90]Apel K,Hirt H.Reactive oxygen species:metabolism,oxidative stress,and signal transduction[J].Annual Review of Plant Biology.2004,55(1):373-399.
[91]Jose N,Janardhanan KK.Antioxidant and antitumour activity of Pleurotus florida[J].Current Science.2000,79(7):941-943.
[92]Morcos NC,Berns M,Henry WL.Phycocyanin:laser activation,cytotoxic effect,and uptake in human atherosclerotic plaque[J].Lasers Surg Med.1988,8(1):7-10.
[93]Galli F,Canestrari F,Bellomo G.Pathophysiology of the oxidative stress and its implication in uremia and dialysis[J].Contrib Nephrol.1999,127:1-31.
[94]Ghiselli A,Serafini M,Maiani G,et al.A fluorescence-based method for measuring total plasma antioxidant capability[J].Free Radical Biology and Medicine.1995,18(1):29-36.
[95]Benedetti S,Benvenuti F,Pagliarani S,et al.Antioxidant properties of a novel phycocyanin extract from the blue-green alga Aphanizomenon flos-aquae[J].Life Sciences.2004,75(19):2353-2362.
[96]Pinero EJE,Bermejo BP,Villar FAM.Antioxidant activity of different fractions of Spirulina platensis protean extract[J].I1 Farmaco.2001,56(5-7):497-500.
[97]魏文志,夏文水,吴玉娟.小球藻蛋白质的分离、纯化及抗氧化特性[J].上海水产大学学报.2007,16(2):135-139。
[98]杨立红,王晓洁,钟旭升等.鱼腥藻藻蓝蛋白的抗氧化作用[J].食品科学.2006,27(12):208-213.
[99]唐玫,王曼,郭宝江.富硒藻蓝蛋白对小鼠免疫功能及抗氧化活性的影响[J].营养学报.2001,23(3):275-278.
[100]陈美珍,张永雨,余杰等.龙须菜藻胆蛋白的分离及其清除自由基作用的初步研究[J].食品科学.2004,25(3):159-162.
[101]陈美珍,余杰,钟秋玲等。龙须菜藻胆蛋白免疫功能和抗氧化作用的研究[J].食品科学.2005,26(9):456.459.
[102]Romay C,GonzAlez R.Phycocyanin is an antioxidant protector of human erythrocytes against lysis by peroxyl radicals[J].Journal of Pharmacy and Pharmacology.2000,52(4):367-368.
[103]Farooq SM,Asokan D,Sakthivel R,et al.Salubrious effect of C-phycocyanin against oxalate-mediated renal cell injury[J].Clinica Chimica Acta.2004,348(1-2):199-205.
[104]Bhat VB,Madyastha KM.C-phycocyanin:a potent peroxyl radical scavenger in vivo and in vitro[J].Biochemical and Biophysical Research Communications.2000,275(1):20-25.
[105]Romay C,Armesto J,Remirez D,et al.Antioxidant and anti-inflammatory properties of C-phycocyanin from blue-green algae[J].Inflammation Research.1998,47(1):36-41.
[106]Romay C,Gonzalez R,Ledon N,et al.C-phycocyanin:a biliprotein with antioxidant,anti-inflammatory and neuroprotective effects[J].Curr Protein Pept Sci.2003,4(3):207-216.
[107]Cherng SC,Cheng SN,Tam A,et al.Anti-inflammatory activity of c-phycocyanin in lipopolysaccharide-stimulated RAW 264.7 macrophages[J].Life Sci.2007,81(19-20):1431-1435.
[108]何立明,郑树.藻蓝蛋白对大肠癌和S180小鼠移植瘤的光敏作用[J].激光医学.1996,6(4):156-157,160.
[109]张成武,曾昭琪.藻蓝蛋白对小鼠粒单系祖细胞生成的影响[J]。中国海洋药物.1996,15(4):25-28.
[110]关燕清,周天鸿.光固定化藻蓝蛋白对内皮细胞生长的影响[J]。功能高分子学报.2003,16(3):373-376.
[111]关燕清,徐和德.光固定化藻蓝蛋白对体外肝癌细胞7402的抑制作用[J].离子交换与吸附.2000,16(6):547-552.
[112]张永雨,陈美珍,余杰等.龙须菜藻胆蛋白抗突变与抗肿瘤作用的研究[J].中国海洋药物.2005,24(3):36-38.
[113]张惟杰.糖复合物生化研究技术(第二版)[M].杭州:浙江大学出版社,1999.11-17
[114]Tandeau MN,Houmard J.Complementary chromatic adaptation:physiological conditions and action spectra[J].Methods Enzymol.1988,167:318-328.
[115]张志良.植物生理学实验指导[M].北京:高等教育出版社,1990。
[116]Chang FH,McClean M.Growth responses ofAlexandrium minutum(Dinophyceae)as a function of three different nitrogen sources and irradiance[J].New Zealand Journal of Marine and Freshwater Research.1997,31(1):1-7.
[117]赵素芬,孙会强.氮源、磷源及重金属离子对绿色巴夫藻生长的影响[J].湛江海洋大学学报.2005,25(1):60-63.
[118]Turpin DH.Effects of inorganic N availability on algal photosynthesis and carbon metabolism[J].Journal of Phycology.1991,27(1):14-20.
[119]Rayburn WR,Kroen WK.Influence of growth status and nutrients on extracellular polysaccharide synthesis by the soil alga Chlamydornonas mexicana(Chlorophyceae)[J].Journal of Phycology.1984,20(2):253-257.
[120]Sloth JK,Eriksen NT.Accumulation of phycocyanin in heterotrophic and mixotrophic cultures of the acidophilic red alga Galdieria sulphuraria[J].Enzyme and Microbial Technology.2006,38(1-2):168-175.
[121]孙辉,徐文华,雷高鹏等.氮、磷、硫对盐生杜氏藻色素积累的影响[J].四川大学学报(自然科学版).2005,42(2):403-407.
[122]蔡恒江,唐学玺,张培玉等.UV-B辐射对青岛大扁藻生长及其某些生理特性的影响[J]。海洋科学进展.2005,23(4):460-465.
[123]Dortch Q,Ahmed SI,Packard TT.Nitrate reductase and glutamate dehydrogenase activities in Skeletonema costatum as measures of nitrogen assimilation rates 1[J].Journal of Plankton Research.2002,1(2):169-186.
[124]李梅燕.西施舌人工育苗饵料藻种的选育[D].福建师范大学硕士学位论文2004:64-65.
[125]Lomas MW.Nitrate reductase and urease enzyme activity in the marine diatom Thalassiosira weissflogii(Bacillariophyceae):interactions among nitrogen substrates[J].Marine Biology.2004,144(1):37-44.
[126]Misra N,Gupta AK.Effect of salinity and different nitrogen sources on the activity of antioxidant enzymes and indole alkaloid content in Catharanthus roseus seedlings[J].Journal of Plant Physiology.2006,163(1):11-18.
[127]章申峰.C-藻蓝蛋白及其亚基的分离纯化和体外抗肿瘤研究[D].浙江大学硕士学位论文.2005:39-41.
[128]Minkova KM,Tchorbadjieva MI,Fournadjieva ST,et al.Purification of C-phycocyanin from Spirulina(Arthrospira)fusiformis[J].Journal of Biotechnology.2003,102:55-59.
[129]欧瑜,董剑,吴梧桐.盐生隐杆藻藻胆蛋白的分离纯化和特性研究[J].药物生物技术.2004,11(1):37-41.
[130]殷钢,李琛.螺旋藻中藻胆蛋白和多糖的分离纯化及产品特性[J].精细化工.1999,16(2):10-13.
[131]Sarada RMGP,Ravishankar GA.Phycocyanin from Spirulina sp influnence of processing of biomass on phycocyanin yield analysis of efficacy of extraction methods and stability studies on phycocyanin[J].Process Biochemistry.1999,34(8):795-801.
[132]范刚,陈德文,潘思铁等.葛仙米藻胆蛋白提取工艺及藻蓝蛋白稳定性研究[J].食品科学.2005,26(9):215-218.
[133]杨芬,张瑞萍,贺玖明等.羟自由基的产生、捕集及检测方法[J].药学学报. 2007,42(7):692-697.
[134]Huycke MM,Abrams Victoria,Moore Danny Ro Enterococcus faecalis produces extracellular superoxide and hydrogen peroxide that damages colonic epithelial cell[J].DNA Carcinogenesis.2002,23(3):529-536.
[135]Wolff SP.Ferrous ion oxidation in presence of ferric ion indicator xylenol orange for measurement of hydroperoxides[J].Mothods in enzymology.1994,233(18):183-189.
[136]周站平,陈秀兰,陈超等.藻胆蛋白脱辅基蛋白对其抗氧化活性的影响[J].海洋科学.2003,27(5):77-80.