磷浓度对海水钝顶螺旋藻生长代谢的影响

doi:10.13560/j.cnki.biotech.bull.1985.2019-0181

摘要/Abstract

摘要： 磷元素对微藻的生长代谢具有重要作用。为了探究磷源浓度调控海水螺旋藻主要代谢产物合成积累的可行性。以改良的Zarrouk海水培养基为基础,通过设置5种K₂HPO₃浓度（0.005-0.04 g/L）,分析不同磷浓度对海水螺旋藻的生长及生理生化指标的影响。结果显示,海水螺旋藻生物质浓度和蛋白质含量随着磷源浓度的升高而增加,0.04 g/L处理组达到最大值,分别为0.48±0.02 g/L和59.23±0.61%DW;总多糖含量随着磷源浓度增加先显著下降,0.005 g/L处理组含量最高（25.96±1.61%DW）;C-藻蓝蛋白、别藻蓝蛋白和总藻胆蛋白含量随着磷源浓度增加,先升后降,0.03 g/L处理组含量最高,分别为14.56±0.99%DW、4.21±0.19%DW和18.77±0.39%DW;叶绿素a与类胡萝卜素含量随着磷源浓度的增加而升高,最高含量分别为1.01±0.01%DW和0.35±0.02%DW;而磷源浓度的变化对总脂含量和脂肪酸组成总体上无显著影响（P>0.05）。低磷源浓度有利于多糖积累,高磷源浓度则促进蛋白质和藻蓝蛋白、叶绿素和类胡萝卜素的合成,通过磷源调节可有效定向诱导海水螺旋藻主要高值化产物的合成积累。

关键词: 海水螺旋藻, 生长, 蛋白质, 多糖, 藻蓝蛋白

Abstract: Phosphorus plays an important role in the growth and metabolism of microalgae. In order to explore the feasibility of regulating the accumulation of major metabolites in seawater Spirulina by adjusting phosphorus concentration,we analyzed the physiological and biochemical effects of different phosphorus concentrations on seawater Spirulina platensis by using K₂HPO₃·3H₂O as phosphorus resource and setting 5 phosphorus concentrations 0.005-0.04 g/L based on seawater Zarrouk medium. The result showed that biomass production and protein content of seawater S. platensis were enhanced along with the increase of phosphorus concentration,and the maximum were 0.48±0.02 g/L and 59.23±0.61%DW,respectively,at the concentration of 0.04 g/L. The total polysaccharide content decreased significantly and then tended to be stable with the increasing of phosphorus concentration,and the maximum was 25.96±1.61%DW at phosphate concentration of 0.005 g/L. The content of C-phycocyanin,allophycocyanin and total phycobiliproteins increased first and then decreased along with the increasing of phosphorus concentration,and the maximum reached at the phosphate concentration of 0.03 g/L by 14.56±0.99%DW,4.21±0.19%DW and 18.77±0.39%DW respectively. The content of chlorophyll a and carotenoids increased along with the increasing of phosphate concentration,with the maximum value of 1.01±0.01%DW and 0.35±0.02%DW,respectively. The variation of phosphorus concentration generally demonstrated no significant effect on total lipids content and fatty acids composition（P>0.05）. The results showed that low phosphorus concentration was beneficial to the accumulation of polysaccharides;while high phosphorus concentration was beneficial to the synthesis of protein and phycocyanin,chlorophyll and carotenoid,thus the synthesis and accumulation of the main high-value products of seawater Spirulina could be effectively adjusted by changing phosphorus concentration.

Key words: seawater Spirulina platensis, growth, protein, polysaccharide, phycocyanin

陈浩, 杨冰洁, 李涛, 吴华莲, 吴后波, 向文洲. 磷浓度对海水钝顶螺旋藻生长代谢的影响[J]. 生物技术通报, 2019, 35(8): 103-110.

CHEN Hao, YANG Bing-jie, LI Tao, WU Hua-lian, WU Hou-bo, XIANG Wen-zhou. Effects of Phosphorus Concentrations on Growth and Metabolism of Seawater Spirulina platensis[J]. Biotechnology Bulletin, 2019, 35(8): 103-110.

参考文献

[1] 李中岳. 螺旋藻的开发利用[J]. 生物学杂志, 1994(1):22-24.
[2] 何善生, 王力, 李健, 等. 螺旋藻研究进展[J]. 食品工业, 2017, 38(12):263-268.
[3] Hayashi K, Hayashi T, Kojima I.A natural sulfated polysaccharide, calcium spirulan, isolated from Spirulina platensis:in vitro and ex vivo evaluation of anti-herpes simplex virus and anti-human immunodeficiency virus activities[J]. Aids Res Hum Retroviruses, 1996, 12(15):1463-1471.
[4] Chaiklahan R, Chirasuwan N, Triratana P, et al.Polysaccharide extraction from Spirulina sp. and its antioxidant capacity[J]. Int J Biol Macromol, 2013, 58:73-78.
[5] Pang QS, Guo BJ, Ruan JH.Enhancement of endonuclease activity and repair DNA synthesis by polysaccharide of Spirulina platensis[J]. Journal of Genetics and Genomics, 1988, 15(5):374-381.
[6] 黄建设, 龙丽娟, 张偲. 海洋天然产物及其生理活性的研究进展[J]. 海洋通报, 2001, 20(4):83-91.
[7] 刘慧慧, 姜良乾, 王玉娟等. 藻蓝蛋白抗肿瘤的应用现状[J]. 肿瘤防治研究, 2018, 45(6):420-424.
[8] 郝帅, 秦玉, 王成涛. 功能食品藻蓝蛋白的生理活性研究进展[J]. 食品与生物技术学报, 2017, 36(12):1233-1240.
[9] 王菲, 佘星星, 孙冰洁, 等. 螺旋藻γ-亚麻酸的提取优化及体外抗氧化活性的研究[J]. 食品工业科技, 2014, 35(19):68-72.
[10] 向文洲, 李涛, 吴华莲, 等. 海水螺旋藻产业发展战略研究;[J]. 广西科学, 2014, 21(6):573-579.
[11] Leema JTM, Kirubagaran R, Vinithkumar NV, et al.High value pigment production from Arthrospira(Spirulina)platensis cultu-red in seawater[J]. Bioresource Technology, 2010, 101(23):9221-9227.
[12] Wu BT, Xiang WZ, Zeng CK.Spirulina cultivation in China[J]. Chinese Journal of Oceanology & Limnology, 1998, 16(1):152-157.
[13] Raghothama KG.Phosphate transport and signaling[J]. Current Opinion in Plant Biology, 2000, 3(3):182-187.
[14] Liang K, Zhang Q, Gu M, et al.Effect of phosphorus on lipid accumulation in freshwater microalga Chlorella sp[J]. Journal of Applied Phycology, 2013, 25(1):311-318.
[15] Minhas AK, Hodgson P, Barrow CJ, et al.A review on the assessment of stress conditions for simultaneous production of microalgal lipids and carotenoids[J]Frontiers in Microbiology, 2016, 7:546.
[16] Baiee, MA, Salman JM. Effect of phosphorus concentration and light intensity on protein content of microalga Chlorella vulgaris[J]Mesopotamia Environmental Journal, 2016, 2(2):75-86.
[17] Brembu T, Alice Mühlroth, Alipanah L, et al.The effects of phosphorus limitation on carbon metabolism in diatoms[J]. Philosophical Transactions of the Royal Society B Biological Sciences, 2017, 372(1728):20160406.
[18] 黄怡, 高保燕, 王飞飞, 等. 不同磷、硫及二氧化碳浓度对标志链带藻生长和碳水化合物积累的影响[J/OL]. 微生物学报, doi:10. 13343/j. cnki. wsxb. 20180459.
[19] Piorreck M.Biomass production, total protein, chlorophylls, lipids and fatty acids of freshwater green and blue-green algae under different nitrogen regimes[J]. Phytochemistry, 1984, 23(2):207-216.
[20] Tedesco MA, Duerr EO.Light, temperature and nitrogen starvation effects on the total lipid and fatty acid content and composition of Spirulina platensis UTEX 1928[J]. Journal of Applied Phycology, 1989, 1(3):201-209.
[21] Ogbonda KH, Aminigo RE, Abu GO.Influence of temperature and pH on biomass production and protein biosynthesis in a putative Spirulina sp[J]. Bioresource Technology, 2007, 98(11):2207-2211.
[22] Markou G, Chatzipavlidis I, Georgakakis D.Effects of phosphorus concentration and light intensity on the biomass composition of Arthrospira(Spirulina)platensis[J]. World Journal of Microbiology and Biotechnology, 2012, 28(8):2661-2670.
[23] Bhakar RN, Brahmdutt B, Pabbi S.Total lipid accumulation and fatty acid profiles of microalga Spirulina under different nitrogen and phosphorus concentrations[J]. Egyptian Journal of Biology, 2014, 16(1):57-62.
[24] Dubois M, Gilles KA, Hamilton JK, et al.Colorimetric method for determination of sugars and related substances[J]. Analytical Chemistry, 1956, 28(3):350-356.
[25] Ahmed AER, Labavitch JM.A simplified method for accurate determination of cell wall uronide content[J]. Journal of Food Biochemistry, 1978, 1(4):5.
[26] Stein JR.Handbook of phycological methods - culture methods and growth measurements[J]. Aquatic Botany, 1973, 10:92-93.
[27] Wan LL, Li AF, Zhang CW.Responses in growth, lipid accumulation, and fatty acid composition of four oleaginous microalgae to different nitrogen sources and concentrations[J]. Chinese Journal of Oceanology & Limnology, 2013, 31(6):1306-1314.
[28] Lichtenthaler HK.Chlorophylls and carotenoids:pigments of photosynthesis[J]. Methods in Enzymology, 1987, 148C(1):350-382.
[29] Khozin-Goldberg I, Shrestha P, Cohen Z.Mobilization of arachidonyl moieties from triacylglycerols into chloroplasticlipids following recovery from nitrogen starvation of the microalga Parietochlorisincisa[J]. Biochim Biophys Acta, 2005, 1738(1):63-71.
[30] Fuentes-Grconewald C, Garccbs E, Rossi S, et al.Use of the dinoflagellate karlodinium veneficum as a sustainable source of biodiesel production[J]. Journal of Industrial Microbiology & Biotechnology, 2009, 36(9):1215-1224.
[31] 董金利, 庄惠如, 占美怜, 等. 氮、磷营养盐对底栖硅藻的生长及生化组成影响[J]. 生物技术, 2010, 20(2):64-67.
[32] 林海, 王源, 李冰. 不同浓度的磷对栅藻生长的影响[J]. 吉林农业, 2018(24):59.
[33] Geider R, Roche JL.Redfield revisited:variability of C:N:P in marine microalgae and its biochemical basis[J]. British Phycological Bulletin, 2002, 37(1):1-17.
[34] 许可. 不同氮磷浓度对普通小球藻生长及光合作用的影响研究[D]. 西安:西安建筑科技大学, 2018.
[35] Gómez-Casati DF, Cortassa S, Aon MA, et al.Ultrasensitive behavior in the synthesis of storage polysaccharides in cyanobacteria[J]. Planta, 2003, 216(6):969-975.
[36] Heldt HW, Chon CJ, Maronde D, et al.Role of orthophosphate and other factors in the regulation of starch formation in leaves and isolated chloroplasts[J]. Plant Physiology, 1977, 59(6):1146-1155.
[37] Berdalet E, Latasa M, Estrada M.Effects of nitrogen and phosphorus starvation on nucleic acid and protein content of Heterocapsa sp[J]. Journal of Plankton Research, 1994, 16(4):303-316.
[38] Dean AP, Estrada B, Nicholson JM, et al.Molecular response of Anabaena flos-aquae to differing concentrations of phosphorus:A combined fourier transform infrared and X-ray microanalytical study[J]. Phycological Research, 2010, 56(3):193-201.
[39] Lynn SG, Kilham SS.Effect of nutrient availability on the biochemical and elemental stoichiometry in the freshwater diatom Stephanodiscus minutulus(Bacillariophyceae)[J]. Journal of Phycology, 2010, 36(3):510-522.