Overview on Efficient Methods for the Determination of Microalgae Lipid Content

doi:10.13560/j.cnki.biotech.bull.1985.2019-0364

Abstract

Abstract: Due to the petroleum reserves diminishing and the concerns of global environmental protection rising,microalgae,as a sustainable and renewable energy source,has been attracted wide attention in recent years. The breeding of microalgae with high oil content is an important step in the development of microalgae bioenergy,and the efficient and rapid determination method of lipids is one of the key technologies in the breeding process. This paper reviews the basic principles and research status of several efficient,accurate and convenient determination methods of lipids,such as Nile red fluorescence spectrometry,BODIPY 505/515 fluorescence spectrometry,copper reagent method,Sudan black B staining method,Fourier transform infrared spectrometry,time-domain nuclear magnetic resonance method,and vis/near-infrared spectrometry. Furthermore,the advantages and methodological limitations of these methods are also compared,aiming at providing a grateful reference for determination of microalgae lipid and concurrently some scientific know-how for subsequent research on the production of microalgae oil.

Key words: microalgae, determination of lipid, efficient method, bioenergy

LI Zhi, ZHOU Qiu-xiang, LI Qiu-ling, LIU Meng-ying, ZHOU Zhi-you, LI Han-guang. Overview on Efficient Methods for the Determination of Microalgae Lipid Content[J]. Biotechnology Bulletin, 2019, 35(12): 189-195.

References

[1] Gatamaneni BL, Orsat V, Lefsrud M.Factors affecting growth of various microalgal species[J]. Environmental Engineering Science, 2018, 35(10):1037-1048.
[2] Faried M, Samer M, Abdelsalam E, et al.Biodiesel production from microalgae:Processes, technologies and recent advancements[J]. Renewable and Sustainable Energy Reviews, 2017, 79:893-913.
[3] Živković SB, Veljković MV, Banković-Ilić IB, et al.Technological, technical, economic, environmental, social, human health risk, toxicological and policy considerations of biodiesel production and use[J]. Renewable and Sustainable Energy Reviews, 2017, 79:222-247.
[4] Japar AS, Takriff MS, Yasin NHM.Harvesting microalgal biomass and lipid extraction for potential biofuel production:A review[J]. Journal of Environmental Chemical Engineering, 2016, 5(1):555-563.
[5] Shomal R, Hisham H, Mlhem A, et al.Simultaneous extraction-reaction process for biodiesel production from microalgae[J]. Energy Reports, 2019, 5:37-40.
[6] Goh BHH, Ong HC, Cheah MY, et al.Sustainability of direct biodiesel synthesis from microalgae biomass:A critical review[J]. Renewable and Sustainable Energy Reviews, 2019, 107:59-74.
[7] 胡文军, 罗玮, 李汉广, 等. 产油微藻筛选和鉴定及其产油性能的研究[J]. 中国生物工程杂志, 2012, 32(12):66-72.
[8] Wei D, Yuan XY, Xiang WZ.Comparison of methods for rapid determination of total lipid content in microalgae[J]. Modern Food Science & Technology, 2014, 30(7):242-246.
[9] Silva MTLD, Reis A.Biodiesel production from microalgae methods for microalgal lipid assessment with emphasis on the use of flow cytometry[M]// Gordon R, Seckbach J. The Science of Algal Fuels. Lisbon:Springer Netherlands, 2012:255-267.
[10] Greenspan P, Fowler SD.Spectrofluorometric studies of the lipid probe, Nile Red[J]. Journal of Lipid Research, 1985, 26(7):781-789.
[11] 蒋晓艳, 王婉玉, 钟敏, 等. 裂殖壶菌胞内油脂尼罗红荧光染色快速检测方法的研究[J]. 中国油脂, 2016, 41(1):51-55.
[12] 曹素娟, 柳科欢, 任玲萱, 等. 快速高通量测定三角褐指藻油脂含量[J]. 四川大学学报:自然科学版, 2018, 55(6):1331-1336.
[13] Orr V, Rehmann L.Improvement of the Nile Red fluorescence assay for determination of total lipid content in microalgae independent of chlorophyll content[J]. Journal of Applied Phycology, 2015, 27(6):1-9.
[14] 丛峰, 孙雪, 徐年军. 几种小球藻油脂含量检测方法的比较及优化[J]. 宁波大学学报:理工版, 2012, 25(1):20-23.
[15] Chen W, Zhang C, Song L, et al.A high throughput Nile Red method for quantitative measurement of neutral lipids in microalgae[J]. Journal of Microbiological Methods, 2015, 77(1):41-47.
[16] Storms ZJ, Cameron E, De LHSH, et al.A simple and rapid protocol for measuring neutral lipids in algal cells using fluorescence[J]. Journal of Visualized Experiments Jove, 2014(87):e51441.
[17] 郝翠翠, 梁成伟, 石蕾. 尼罗红和BODIPY 荧光染料在微藻油脂含量测定中的应用[J]. 生物学杂志, 2017, 34(1):70-74.
[18] Koreivienė J.Microalgae lipid staining with fluorescent BODIPY dye[J]. Methods in Molecular Biology, 2017, doi:10.1007/7651.2017-101.
[19] Rumin J, Bonnefond H, Saintjean B, et al.The use of fluorescent Nile red and BODIPY for lipid measurement in microalgae[J]. Biotechnology for Biofuels, 2015, 8(1):42.
[20] 石玉新, 穆迪, 武洪庆, 等. 微藻油脂含量的几种快速测定方法[J]. 安徽农业科学, 2012, 40(21):11067-11069.
[21] Wawrik B, Harriman BH.Rapid, colorimetric quantification of lipid from algal cultures[J]. Journal of Microbiological Methods, 2010, 80(3):262-266.
[22] Subramaniam HN, Chaubal KA.Evaluation of intracellular lipids by standardized staining with a Sudan Black B fraction[J]. Journal of Biochemical & Biophysical Methods, 1990, 21(1):9-16.
[23] Thakur MS, Prapulla SG, Karanth NG.Estimation of intracellular lipids by the measurement of absorbance of yeast cells stained with Sudan Black B[J]. Enzyme and Microbial Technology, 1989, 11(4):252-254.
[24] Ren J, Lang XY, Liu ZY, et al.Rapidly determinating relative lipid level of Chlorella vulgaris sp. by three dyeing methods[J]. Journal of Agricultural Biotechnology, 2015, 23(7):967-972.
[25] 金雪洁, 巩三强, 夏金兰, 等. 苏丹黑B染色法原位检测2株淡水小球藻胞内油脂累积[J]. 中南大学学报:自然科学版, 2014, 45(8):2547-2552.
[26] 彭悦, 刘玉莹, 祁艳霞, 等. 微藻油脂含量不同测定方法的比较研究[J]. 大连海洋大学学报, 2015, 30(5):455-460.
[27] 杜秀秀, 房志家, 陈忠翔, 等. 酵母中性脂快速检测及积累动态分析[J]. 生物技术通报, 2014(8):208-214.
[28] 李贵丽, 韩志萍, 魏晓奕, 等. 能源微藻油脂积累及油脂含量检测技术研究进展[J]. 广东农业科学, 2018, 45(10):134-142.
[29] Giordano M, Kansiz M, Heraud P, et al.Fourier transform infrared spectroscopy as a novel tool to investigate changes in intracellular macromolecular pools in the marine microalga Chaetoceros muellerii(Bacillariophyceae)[J]. Journal of Phycology, 2010, 37(2):271-279.
[30] Jyoti S, Smitas K, Narsi RB, et al.Screening and enrichment of high lipid producing microalgal consortia[J]. Journal of Photochemistry and Photobiology B:Biology, 2019, 192(3):8-12.
[31] Dean AP, Sigee DC, Estrada B, et al.Using FTIR spectroscopy for rapid determination of lipid accumulation in response to nitrogen limitation in freshwater microalgae[J]. Bioresource Technology, 2010, 101(12):4499-4507.
[32] 陈小妹, 张敬键, 吕雪娟, 等. FTIR法快速测定产油微藻细胞内生化组分[J]. 生物技术, 2013, 23(2):60-65.
[33] Sarpal AS, Teixeira CMLL, Silva PRM, et al.NMR techniques for determination of lipid content in microalgal biomass and their use in monitoring the cultivation with biodiesel potential[J]. Applied Microbiology and Biotechnology, 2016, 100(5):2471-2485.
[34] 曹玉坡, 刘义军, 黄晖, 等. 应用低场核磁共振技术测定腰果仁中的含油量[J]. 食品与发酵科技, 2016, 52(5):71-74.
[35] Gao C, Wei X, Zhang Y, et al.Rapid quantitation of lipid in microalgae by time-domain nuclear magnetic resonance[J]. Journal of Microbiological Methods, 2008, 75(3):437-440.
[36] 刘婷婷, 王涛, 杨翼, 等. 低场核磁共振技术快速检测小球藻油脂含量及其在高通量选育中的应用[J]. 生物工程学报, 2016, 32(3):385-396.
[37] 刘京华, 陈军, 秦松, 等. 红外光谱在微藻领域的应用研究进展[J]. 光谱学与光谱分析, 2019, 39(1):79-86.
[38] 魏萱, 蒋璐璐, 赵艳茹, 等. 小球藻生长过程脂肪含量动态变化快速无损检测方法研究[J]. 光谱学与光谱分析, 2016, 36(5):1352-1357.
[39] Brown MR, Frampton DMF, Dunstan GA, et al.Assessing near-infrared reflectance spectroscopy for the rapid detection of lipid and biomass in microalgae cultures[J]. Journal of Applied Phycology, 2014, 26(1):191-198.