LI Jie, HUANG Hui, XIANG Huan, CEN Jianwei, WEI Ya, ZHAO Yongqiang, HAO Shuxian, LI Laihao. Extraction process of intestinal lipid droplets and lipid droplet-related proteins from tilapia[J]. South China Fisheries Science, 2023, 19(3): 119-128. DOI: 10.12131/20220285
Citation: LI Jie, HUANG Hui, XIANG Huan, CEN Jianwei, WEI Ya, ZHAO Yongqiang, HAO Shuxian, LI Laihao. Extraction process of intestinal lipid droplets and lipid droplet-related proteins from tilapia[J]. South China Fisheries Science, 2023, 19(3): 119-128. DOI: 10.12131/20220285

Extraction process of intestinal lipid droplets and lipid droplet-related proteins from tilapia

More Information
  • Received Date: October 31, 2022
  • Revised Date: December 17, 2022
  • Accepted Date: December 19, 2022
  • Available Online: February 18, 2023
  • Lipids in animals and plants exist in the form of lipid droplet (LD). It is very important to master the purification methods of lipid droplet and lipid droplet related protein (LDRP) for understanding the lipid changes and influence mechanisms of fish, and seeking appropriate regulatory approaches. In this paper, the lipid droplet and LDRP were separated and purified through centrifugation, resuspension and solubilization by using Oreochromis niloticus intestinal tissues as raw material. The results show that the intestinal adipose tissues were effectively removed by three times of centrifugation and three times of resuspension to obtain the pure lipid droplet. Although the concentration of LDRP protein extracted by solubilization was higher, the electrophoretic bands showed an obvious trailing phenomenon, indicating that the effect of lipid removal was poor, which affected the purity of LDRP extraction. Delipidation and solubilization need to be combined with four groups of organic solvents to obtain the LDRP extracted samples with high purity and good re-solubilization effects. The effects of centrifugation drying method and lipid droplet freezing on LDRP solubilization were not significantly different.
  • [1]
    林剑军, 赵文红, 刘巧瑜, 等. 脂质水解氧化对干腌鱼制品风味影响的研究进展[J]. 食品工业, 2021, 42(9): 206-210.
    [2]
    OKPALA C O R, BONO G. Lipid autoxidation in fishery products postharvest: is cold to freezing oxygen-free packaged storage the better remedy?[J]. Lipid Technol, 2016, 28(7): 122-125. doi: 10.1002/lite.201600034
    [3]
    韩迎雪, 林婉玲, 杨少玲, 等. 15种淡水鱼肌肉脂肪含量及脂肪酸组成分析[J]. 食品工业科技, 2018, 39(20): 217-222.
    [4]
    柯欢, 李慧, 熊伟, 等. 白乌鱼与罗非鱼肉的营养物质比较分析[J]. 农产品加工, 2020(1): 47-49.
    [5]
    庞永佳, 翟桂影, 李瑞, 等. 脂滴包被蛋白的结构、功能及其与脂类疾病关联的研究进展[J]. 中国畜牧杂志, 2021, 57(6): 59-67.
    [6]
    彭玥晗, 徐磊, 许金蔓, 等. 脂滴包被蛋白基因对猪肌肉发育的影响[J]. 畜牧兽医杂志, 2019, 38(3): 28-30.
    [7]
    SHIVAIAH K K, SUSANTO F A, DEVADASU E, et al. Plastoglobule lipid droplet isolation from plant leaf tissue and cyanobacteria[J]. J Vis Exp, 2022: 188. doi: 10.3791/64515
    [8]
    罗玉珍, 韩海军, 陈敏, 等. 脂滴包被蛋白Plin5参与调节脂代谢和线粒体功能[J]. 生命的化学, 2020, 40(8): 1279-1288.
    [9]
    SINGH A, SEN P. Lipid droplet: a functionally active organelle in monocyte to macrophage differentiation and its inflammatory properties[J]. Biochim Biophys Acta Mol Cell Biol Lipids, 2021, 1866(10): 158981.
    [10]
    WILFLING F, WANG H J, HAAS J T, et al. Triacylglycerol synthesis enzymes mediate lipid droplet growth by relocalizing from the ER to lipid eroplets[J]. Dev Cell, 2013, 24(4): 384-399.
    [11]
    DEJGAARD S Y, PRESLEY J F. New method for quantitation of lipid droplet volume from light microscopic images with an application to determination of pat protein density on the droplet surface[J]. J. Histochem Cytochem, 2018, 66(6): 447-465. doi: 10.1369/0022155417753573
    [12]
    徐冲, 何金汗, 徐国恒. 脂滴包被蛋白 (perilipin) 调控脂肪分解[J]. 生理科学进展, 2006(3): 221-224.
    [13]
    刘梅芳, 徐国恒. PAT家族蛋白在细胞内脂滴代谢过程中的作用[J]. 生理科学进展, 2006(2): 103-107.
    [14]
    YAN Z H, ZHAO L P, KONG X Z, et al. Behaviors of particle size and bound proteins of oil bodies in soymilk processing[J]. Food Chem, 2016, 1866(10): 881-890.
    [15]
    钟艳花, 林重, 钟映芹, 等. 厚朴酚对脂肪酸诱导HepG2细胞脂质积蓄的作用及机制研究[J]. 山西中医药大学学报, 2021, 22(3): 171-177.
    [16]
    MURPHY D. The biogenesis and functions of lipid bodies in animals, plants and microorganisms[J]. Prog Lipid Res, 2001, 40(5): 325-438. doi: 10.1016/S0163-7827(01)00013-3
    [17]
    BHATLA S C, VANDANA S, KAUSHIK V. Recent developments in the localization of oil body-associated signaling molecules during lipolysis in oilseeds[J]. Plant Signal Behav, 2009, 4(3): 176-182. doi: 10.4161/psb.4.3.7799
    [18]
    刘芳, 邢颖, 相金柱, 等. 猪早期孤雌胚胎脂滴含量和脂肪酸代谢相关基因表达的初步研究[J]. 内蒙古大学学报(自然科学版), 2019, 50(4): 387-396.
    [19]
    ABDULSAMIE H, MOUHNAD S, MURPHY D J. Functional involvement of caleosin/peroxygenase PdPXG4 in the accumulation of date palm leaf lipid droplets after exposure to dioxins[J]. Environ Pollut, 2021, 281: 116966. doi: 10.1016/j.envpol.2021.116966
    [20]
    MARTA B, ALBERT P. Eukaryotic lipid droplets: metabolic hubs, and immune first responders[J]. Trends Endocrinol, 2022, 33(3): 218-229. doi: 10.1016/j.tem.2021.12.006
    [21]
    CHITRAJU C, FISCHER A W, FARESE R V, et al. Lipid droplets in brown adipose tissue are dispensable for cold-induced thermogenesis[J]. Cell Rep, 2020, 33(5): 108348. doi: 10.1016/j.celrep.2020.108348
    [22]
    YOVANNY I, RUBÉN F S, TOMÁS C, et al. Lipid droplet isolation from arabidopsis thaliana leaves[J]. Bio Protoc, 2020, 10(24): e3867. doi: 10.21769/BioProtoc.3867
    [23]
    TIAN J J, ZHANG J M, YU E M, et al. Identification and analysis of lipid droplet-related proteome in the adipose tissue of grass carp (Ctenopharyngodon idella) under fed and starved conditions[J]. Comp Biochem Physiol D, 2020, 36: 100710.
    [24]
    闫峻, 马衍旋, 穆淑琴, 等. 猪骨骼肌肌内脂滴分离和纯化方法的探究[J]. 动物营养学报, 2019, 31(1): 266-273.
    [25]
    DING Y F, ZHANG S Y, YANG L, et al. Isolating lipid droplets from multiple species[J]. Nat Protoc, 2013, 8(1): 43-51. doi: 10.1038/nprot.2012.142
    [26]
    崔留娟. 棕色脂肪组织脂滴和线粒体相互作用的研究[D]. 合肥: 中国科学技术大学, 2019: 31.
    [27]
    MIRZA A H, CUI L J, ZHANG S Y, et al. Comparative proteomics reveals that lipid droplet-anchored mitochondria are more sensitive to cold in brown adipocytes[J]. Biochim Biophys Acta Mol Cell Biol Lipids, 2021, 1866(10): 158992.
    [28]
    BRASAEMLE D L, WOLINS N E. Isolation of lipid droplets from cells by density gradient centrifugation[J]. Curr Protoc Cell Biol, 2016, 72: 3.15.1-3.15.13. doi: 10.1002/cpcb.10
    [29]
    郭玉. 骨骼肌与脂肪组织细胞中脂滴表面蛋白的差异分析与鉴定[D]. 武汉: 华中农业大学, 2019: 19-21.
    [30]
    HUANG T, BAMIGBABE A T, XU S M, et al. Identification of noncoding RNA-encoded proteins on lipid droplets[J]. Sci Bull, 2021, 66(4): 314-318. doi: 10.1016/j.scib.2020.09.022
    [31]
    王雅文. 罗非鱼脂肪组织和脂肪细胞的代谢生理特性研究[D]. 上海: 华东师范大学, 2017: 27.
    [32]
    林婉玲, 关熔, 曾庆孝, 等. 彩鲷和普通罗非鱼不同部位营养及质构特性的研究[J]. 现代食品科技, 2011, 27(1): 16-21, 49.
    [33]
    王来亮, 周芳芳, 黄璐璐, 等. 应用iTRAQ技术联合PRM验证技术筛选肝硬化患者急性肾损伤的早期预测生物标志物[J]. 现代实用医学, 2021, 33(4): 440-442.
    [34]
    李科, 魏义勇, 刘云, 等. Nycodenz密度梯度离心法提取及纯化大鼠心肌线粒体[J]. 遵义医学院学报, 2016, 39(5): 525-528.
    [35]
    黄靖娴. 超速离心机技术原理与操作维护注意事项[J]. 科技创新与应用, 2018, 36: 130-132.
    [36]
    JOLIVET P, BOULARD C, BELLAMY A, et al. Oil body proteins sequentially accumulate throughout seed development in Brassica napus[J]. J Plant Physiol, 2011, 168(17): 2015-2020. doi: 10.1016/j.jplph.2011.06.007
    [37]
    DING Y F, LI Y, ZHANG S Y, et al. Identification of the major functional proteins of prokaryotic lipid droplets[J]. J Lipid Res, 2012, 53(3): 399-411.
    [38]
    蔡立鹏, 康娜, 谭金玲, 等. 脂滴与胞内细胞器互作研究进展[J]. 中国细胞生物学学报, 2022, 44(5): 964-972.
    [39]
    张来福, 尹云厚. 脂滴的生物物理学和细胞生物学研究[J]. 现代农业科技, 2019, 11: 90-93.
    [40]
    王丽娜, 权春善, 许永斌, 等. 利用表面活性剂介导的方法制备 AgrC 蛋白脂质体[J]. 化学学报, 2014, 72(2): 233-240.
    [41]
    林爱华, 孙小梅, 李步海. 表面活性剂对蛋白质变性作用的研究[J]. 中南民族大学学报(自然科学版) 2002, 21(1): 21-24.
    [42]
    杨小刚. 原油乳状液冻融稳定性的研究[D]. 天津: 天津大学, 2009: 15-18.
    [43]
    王亚娟, 范亚苇, 李静, 等. 不同辅助法对冻融破乳提取油茶籽油的影响[J/OL]. 中国油脂, 2022. https://kns.cnki.net/kcms/detail/61.1099.ts.20220407.0911.001.html.
    [44]
    卢浩, 刘懿谦, 代品一, 等. 油水强化分离技术[J]. 化工进展, 2020, 39(12): 4954-4962.
    [45]
    叶元土. 提升淡水养殖鱼类食用质量的技术集成与创新[J]. 饲料工业, 2022, 43(10): 1-11.
    [46]
    罗强, 闫峻, 李宁, 等. 动物脂滴结构及功能研究进展[J]. 家畜生态学报, 2022, 43(2): 1-7. doi: 10.3969/j.issn.1673-1182.2022.02.001
    [47]
    黎渊弘, 罗艳萍, 廖共山. 短尾蝮蛇毒磷脂结合抗凝蛋白的酶学性质研究[J]. 重庆医科大学学报, 2010, 35(11): 1639-1641.
    [48]
    田鑫, 汪之和, 施文正, 等. 南极磷虾体内胰蛋白酶的纯化及性质研究[J]. 上海海洋大学学报, 2014, 23(5): 741-747.
    [49]
    PERBELLINI F, LIU A K L, WATSON S A, et al. Free-of-Acrylamide SDS-based Tissue Clearing (FASTClear) for three dimensional visualization of myocardial tissue[J]. Sci Rep, 2017, 7(1): 5188. doi: 10.1038/s41598-017-05406-w
    [50]
    LAI M J, LI X W, LI J, et al. Improved clearing of lipid droplet-rich tissues for three-dimensional structural elucidation[J]. Acta Biochim Biophys Sin (Shanghai), 2017, 49(5): 465-467. doi: 10.1093/abbs/gmx018
    [51]
    岳红卫, 韩富亮. 酿酒酵母蛋白提取工艺优化[J]. 食品工业科技, 2015, 16: 304-306.
    [52]
    罗志刚, 杨欢, 齐亮. 生物表面活性剂鼠李糖脂性质的研究[J]. 华南理工大学学报(自然科学版), 2022, 50(1): 30-37.
    [53]
    柳青海, 张唐伟, 李天才. 肝蛋白提取纯化及应用研究进展[J]. 贵州农业科学, 2011, 39(2): 161-164.
    [54]
    王世宽, 于海光, 许艳丽. 叶蛋白提取方法的比较及发酵酸法的应用前景[J]. 生物技术, 2010, 20(1): 94-96.
  • Related Articles

    [1]LYU Junlin, CHEN Zuozhi, LI Bilong, CAI Runji, GAO Yuefang. Research on fish recognition based on multi-stage feature extraction learning[J]. South China Fisheries Science, 2024, 20(1): 99-109. DOI: 10.12131/20230197
    [2]LI Hongting, ZHANG Shuai, ZOU Keshu, CHEN Zuozhi, CHEN Xiaolei, JIANG Peiwen, CAO Yiting, LI Min. Establishment and optimization of environmental DNA extraction method from water of Pearl River Estuary[J]. South China Fisheries Science, 2022, 18(3): 30-37. DOI: 10.12131/20210304
    [3]MIAO Junkui, ZHANG Yating, ZHANG Shengguo, ZHENG Zhihong, WANG Zhipeng, WANG Haiying. Comparative research of plant hormones and alginate oligosaccharides in seaweed extracts by different processing methods[J]. South China Fisheries Science, 2022, 18(2): 142-149. DOI: 10.12131/20210287
    [4]PEI Ruonan, ZHAI Honglei, QI Bo, YANG Xianqing. Optimization of multi-enzymatic extraction of polysaccharide from Gelidium amansii by response surface methodology[J]. South China Fisheries Science, 2019, 15(6): 88-95. DOI: 10.12131/20190081
    [5]WANG Yun, LI Zheng, DUAN Yafei, WANG Jun, HUANG Zhong, LIN Heizhao. Effect of dietary Rhodiola rosea extract on antioxidant system and anti-low salinity stress of Litopenaeus vannamei[J]. South China Fisheries Science, 2018, 14(1): 9-19. DOI: 10.3969/j.issn.2095-0780.2018.01.002
    [6]ZHANG Wan, WU Yanyan, LI Laihao, WANG Jinxu, YANG Shaoling, RONG Hui. Optimization of extraction of antiultraviolet radiation compounds from Gloiopeltis furcata[J]. South China Fisheries Science, 2016, 12(2): 102-109. DOI: 10.3969/j.issn.2095-0780.2016.02.015
    [7]WANG Meng, LI Chun-hou, DAI Ming. The research progress and development trend on extraction of biofuel from marine microalgae[J]. South China Fisheries Science, 2009, 5(2): 74-80. DOI: 10.3969/j.issn.1673-2227.2009.02.013
    [8]YE Xiaoyan, CENG Shaokui, YU Wenguo, WU Wenlong, CENG Xianggen, HUANG Liming. Study on nutrient components and the extracting condition of the skin gelatin of tilapia[J]. South China Fisheries Science, 2008, 4(5): 55-60.
    [9]XU Jing. Study on the extraction of natural astaxanthin by orthogonal test[J]. South China Fisheries Science, 2008, 4(2): 69-74.
    [10]FAN Wujiang, WANG Xiaoqing, YANG Pinhong, XIE Chunhua. Comparison of genomic DNA extraction of Aristichthys nobilis[J]. South China Fisheries Science, 2007, 3(1): 44-47.
  • Cited by

    Periodical cited type(1)

    1. 郝淑贤,韦丽娜,魏涯,黄卉,相欢,赵永强,岑剑伟,王迪,李来好. 发热包传热效果及其对冻干鱼肉复水品质的影响. 广东海洋大学学报. 2024(04): 139-146 .

    Other cited types(0)

Catalog

    Article views (723) PDF downloads (44) Cited by(1)
    Related

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return