There is a lack of simple and efficient separation method of sulfated fucan and chondroitin sulfate in sea cucumber. This study intends to establish a method based on polyethylene glycol (PEG)-salt aqueous two-phase system for the highly selective separation of sulfated fucan and chondroitin sulfate. Using the distribution coefficient and recovery rate as parameters, the effect of different salt types, PEG molecular weight, pH, concentration of PEG and K2HPO4 on the partition behavior of sulfated fucan and chondroitin sulfate from Acaudina molpadioides was discussed. The sulfated fucan and chondroitin sulfate could be separated by the PEG-K2HPO4 aqueous two-phase system. Sulfated fucan was majorly presented in the upper phase, while chondroitin sulfate distributed in the lower phase. Under the most favorable partition conditions with 15% PEG 6000 and 10% K2HPO4, the partition coefficient and recovery rate of sulfated fucan were achieved 6.36 and (86.92±0.97)%. Meanwhile, the partition coefficient and recovery rate of chondroitin sulfate were achieved 16.53 and (106.47±1.84)%, respectively. The aqueous two-phase separation method established in this study could implement the effective separation of sulfated fucan and chondroitin sulfate from Acaudina molpadioides. This method can be applied to the separation of sulfated fucan and chondroitin sulfate of other sea cucumbers species, which has a good application prospect.
SONG Lin
,
SONG Xiao
,
XUE Changhu
,
LI Jiajing
,
LIU Guanchen
,
CHEN Guangning
,
LI Jiayu
,
LIU Jinqiu
,
CHANG Yaoguang
. Study on the PEG-salt aqueous two-phase method for the separation of sulfated fucan and chondroitin sulfate in Acaudina molpadioides[J]. Food and Fermentation Industries, 2023
, 49(19)
: 183
-189
.
DOI: 10.13995/j.cnki.11-1802/ts.034265
[1] 王静杰, 钟强, 董春晖, 等.海参多糖生物学活性及其作用机制研究进展[J].食品科学, 2021, 42(23):370-380.
WANG J J, ZHONG Q, DONG C H, et al.Recent progress in the biological activity and mechanism of action of sea cucumber polysaccharides[J].Food Science, 2021, 42(23):370-380.
[2] 张帅. 刺参岩藻聚糖硫酸酯对RAW264.7细胞炎症和自噬的影响[D].杨凌:西北农林科技大学, 2021.
ZHANG S.Effects of fucoidan from Apostichopus japonicus on inflammation and autophagy in RAW264.7 cells[D].Yangling:Northwest A & F University, 2021.
[3] 尹嘉玉. 海参岩藻聚糖改善LPS诱导的C57BL/6J小鼠急性肝损伤的机制[D].哈尔滨:哈尔滨商业大学, 2020.
YIN J Y.The mechanisms of the fucoidan from sea cucumber Apostichopus japonicus on attenuating lipopolysaccharide-challenged liver injury in C57BL/6J mice[D].Harbin:Harbin University of Commerce, 2020.
[4] 李启冬. 海参肠壁硫酸软骨素的结构解析和活性研究[D].哈尔滨:哈尔滨工业大学, 2021.
LI Q D.Structural elucidation and activity studies of fucosylated chondroitin sulfate from intestinal wall of sea cucumber Stichopus japonicas[D].Harbin:Harbin Institute of Technology, 2021.
[5] LI Y, LI M, XU B J, et al.The Current status and future perspective in combination of the processing technologies of sulfated polysaccharides from sea cucumbers:A comprehensive review[J].Journal of Functional Foods, 2021, 87:104744.
[6] LIU X X, ZHANG Z X, MAO H, et al.Characterization of the hydrolysis kinetics of fucosylated glycosaminoglycan in mild acid and structures of the resulting oligosaccharides[J].Marine Drugs, 2020, 18(6):286.
[7] YANG W J, CAI Y, YIN R H, et al.Structural analysis and anticoagulant activities of two sulfated polysaccharides from the sea cucumber Holothuria coluber[J].International Journal of Biological Macromolecules, 2018, 115:1055-1062.
[8] SHI D L, QI J H, ZHANG H W, et al.Comparison of hydrothermal depolymerization and oligosaccharide profile of fucoidan and fucosylated chondroitin sulfate from Holothuria floridana[J].International Journal of Biological Macromolecules, 2019, 132:738-747.
[9] MA Y, GAO N, ZUO Z C, et al.Five distinct fucan sulfates from sea cucumber Pattalus mollis:Purification, structural characterization and anticoagulant activities[J].International Journal of Biological Macromolecules, 2021, 186:535-543.
[10] DONG X D, PAN R J, DENG X Y, et al.Separation, purification, anticoagulant activity and preliminary structural characterization of two sulfated polysaccharides from sea cucumber Acaudina molpadioidea and Holothuria nobilis[J].Process Biochemistry, 2014, 49(8):1352-1361.
[11] NAZER B, DEHGHANI M R, GOLIAEI B.Plasmid DNA affinity partitioning using polyethylene glycol - sodium sulfate aqueous two-phase systems[J].Journal of Chromatography.B, Analytical Technologies in the Biomedical and Life Sciences, 2017, 1044-1045:112-119.
[12] JOSHI P U, TURPEINEN D G, WEISS M, et al.Tie line framework to optimize non-enveloped virus recovery in aqueous two-phase systems[J].Journal of Chromatography.B, Analytical Technologies in the Biomedical and Life Sciences, 2019, 1126-1127:121744.
[13] MARCHEL M, JOÃO K G, MARRUCHO I M.On the use of ionic liquids as adjuvants in PEG-(NH4)2SO4 aqueous biphasic systems:Phase diagrams behavior and the effect of IL concentration on myoglobin partition[J].Separation and Purification Technology, 2019, 210:710-718.
[14] ZHU L N, LU Y, SUN Z, et al.The application of an aqueous two-phase system combined with ultrasonic cell disruption extraction and HPLC in the simultaneous separation and analysis of solanine and Solanum nigrum polysaccharide from Solanum nigrum unripe fruit[J].Food Chemistry, 2020, 304:125383.
[15] OSLOOB M, ROOSTA A.Experimental study of choline chloride and K2HPO4 aqueous two-phase system, and its application in the partitioning of penicillin G[J].Journal of Molecular Liquids, 2019, 279:171-176.
[16] VIEIRA A W, DA CRUZ SILVA K, MAGESTE A B, et al.Lycopene partition in new aqueous two-phase systems[J].Journal of Molecular Liquids, 2021, 339:116755.
[17] 郝晓棠. 双水相萃取技术分离纯化水中腺病毒和流感病毒的研究[D].广州:广东药科大学, 2021.
HAO X T.Separation and purification of adenovirus and influenza virus from water by aqueous two phase extraction technology[D].Guangzhou:Guangdong Pharmaceutical University, 2021.
[18] RAGHAVA RAO J, NAIR B U.Novel approach towards recovery of glycosaminoglycans from tannery wastewater[J].Bioresource Technology, 2011, 102(2):872-878.
[19] XING J M, LI F F.Purification of aloe polysaccharides by using aqueous two-phase extraction with desalination[J].Natural Product Research, 2009, 23(15):1424-1430.
[20] 巫永华, 陆文静, 刘梦虎, 等.响应面优化超声波辅助双水相提取牛蒡多糖及抗氧化分析[J].食品与发酵工业, 2020, 46(5):215-223.
WU Y H, LU W J, LIU M H, et al.Optimization of ultrasonic-assisted aqueous two-phase extraction of burdock polysaccharide by response surface design and its antioxidant activities[J].Food and Fermentation Industries, 2020, 46(5):215-223.
[21] YANG D D, LIN F D, HUANG Y Y, et al.Separation, purification, structural analysis and immune-enhancing activity of sulfated polysaccharide isolated from sea cucumber viscera[J].International Journal of Biological Macromolecules, 2020, 155:1003-1018.
[22] 续晓琪, 薛长湖, 张翠玉, 等.次甲基蓝显色法定量海参硫酸软骨素[J].食品科学, 2013, 34(22):246-249.
XU X Q, XUE C H, ZHANG C Y, et al.Determination of chondroitin sulfate from sea cucumber by methylene blue colorimetry[J].Food Science, 2013, 34(22):246-249.
[23] VIJAYARAGAVAN K S, ZAHID A, YOUNG J W, et al.Separation of porcine parvovirus from bovine serum albumin using PEG-salt aqueous two-phase system[J].Journal of Chromatography B, 2014, 967:118-126.
[24] ALBERTSSON P Å.Partition of cell particles and macromolecules in polymer two-phase systems[J].Advances in Protein Chemistry, 1970, 24:309-341.
[25] MOHAMADI H S, OMIDINIA E, DINARVAND R.Evaluation of recombinant phenylalanine dehydrogenase behavior in aqueous two-phase partitioning[J].Process Biochemistry, 2007, 42(9):1296-1301.
[26] ANDERSSON E, HAHN-HÄGERDAL B.Bioconversions in aqueous two-phase systems[J].Enzyme and Microbial Technology, 1990, 12(4):242-254.
