聚合物/磷酸盐双水相分离乳清分离蛋白中<i>α</i>-乳白蛋白和<i>β</i>-乳球蛋白条件的优化

引用本文: 张鹤, 姜彬, 冯志彪, 屈玉霄, 李璇. 聚合物/磷酸盐双水相分离乳清分离蛋白中α-乳白蛋白和β-乳球蛋白条件的优化[J]. 分析化学, 2016, 44(5): 754-759. doi: 10.11895/j.issn.0253-3820.150800 shu

Citation: ZHANG He, JIANG Bin, FENG Zhi-Biao, QU Yu-Xiao, LI Xuan. Separation of α-Lactalbumin and β-Lactoglobulin in Whey Protein Isolate by Aqueous Two-phase System of Polymer/Phosphate[J]. Chinese Journal of Analytical Chemistry, 2016, 44(5): 754-759. doi: 10.11895/j.issn.0253-3820.150800 shu

聚合物/磷酸盐双水相分离乳清分离蛋白中α-乳白蛋白和β-乳球蛋白条件的优化

东北农业大学理学院应用化学系, 哈尔滨 150030
基金项目:
本文系国家自然科学基金(No.31201366)和黑龙江省教育厅面上项目(No.12531007)资助

摘要: 建立了基于聚(乙二醇-ran-丙二醇)单丁基醚(聚合物)与磷酸盐、氯化钠的双水相体系对乳清分离蛋白(WPI)中的α-乳白蛋白(α-LA)和β-乳球蛋白(β-LG)进行分离,并对其分离条件进行了优化。系统地研究了双水相体系pH值、聚合物与KH₂PO₄溶液体积比、NaCl添加量和WPI浓度对α-LA和β-LG的分离效果的影响,采用液相色谱法对α-LA和β-LG的分离效果进行评价。结果表明,聚合物/KH₂PO₄双水相体系pH=4.0,40%(m/m)聚合物与15.5%(m/m)KH₂PO₄的体积比为4 mL:4 mL,NaCl添加量为0.40 g/10 mL,WPI浓度为1 mg/mL时,α-LA和β-LG分离效果最好,上相中α-LA的萃取率为98.2%,下相中β-LG的萃取率为96.6%。

关键词:

English

Separation of α-Lactalbumin and β-Lactoglobulin in Whey Protein Isolate by Aqueous Two-phase System of Polymer/Phosphate

Department of Applied Chemistry, Northeast Agricultural University, Harbin 150030, China
Received Date: 12 October 2015
Revised Date: 12 January 2016
Fund Project:
This work was supported by the National Natural Science Foundation of China (No. 31201366)

Abstract: Poly (ethylene glycol-ran-propylene glycol) monobutyl ether/phosphate aqueous two-phase system (ATPS) coupled with high performance liquid chromatography (HPLC) was developed for the separation of α-lactalbumin (α-LA) and β-lactoglobulin (β-LG) in whey protein isolate (WPI). The main parameters relating to this method were investigated and optimized. The effects of pH value, volume ratio of polymer and KH₂PO₄ solutions, concentration of NaCl and WPI on the partition of α-LA and β-LG in WPI were investigated. The results showed that efficient separation of both proteins in WPI could be achieved by using the ATPS with 4 mL poly (ethylene glycol-ran-propylene glycol) monobutyl ether solution (40%, m/m) and 4 mL KH₂PO₄ solution (15.5%, m/m), 0.40 g/10 mL NaCl, while the concentration of WPI was 1 mg/mL, at pH 4.0. Under the optimized conditions, α-LA was separated to the upper phase while β-LG to the bottom phase with the yield of 98.2% and 96.6% respectivelon

Key words:

Aqueous two-phase system
/ Whey protein isolate
/ Poly (ethylene glycol-ran-propylene glycol) monobutyl ether
/ Separation

1. [1]
  Boutin C, Giroux H J, Paquin P, Britten M. Int. Dairy J., 2007,17(6):696-703
2. [2]
  Chatterton D E W, Smithers G, Roupas P, Brodkorb A. Int. Dairy J., 2006,16(11):1229-1240
3. [3]
  Siso M I G. Bioresource Technol., 2006,57(1):1-11
4. [4]
  Alcântara L A P, Minima L A,Minima V P R,Bonomo R C F, Da Silva L H M, Da Silve M C H. J. Chromatogr. B, 2011,879(21):1881-1885
5. [5]
  Jara F, Pilosof A-M R. Food Hydrocoll., 2011,25(3):374-380
6. [6]
  Alcântara L A P, Amaral I V, Bonomo R C F, Da Silva L H M, Da Silva M C H, Minim V P R, Minim L A. Food Bioprod. Process, 2014,92(4):409-415
7. [7]
  Kalaivani S, Regupathi I. J. Sep. Sci. Technol., 2015,50(6):833-840
8. [8]
  Kalaivani S, Regupathi I. Sep. Purif. Technol., 2015, 146(26):301-310
9. [9]
  Chen B, Han J, Wang Y, Sheng C Z, Liu Y, Zhang G C, Yan Y S. Food Chem., 2014,148(1):105-111
10. [10]
  Dembczyński R, Białas W, Jankowski T. Food Bioprod. Process, 2013,91(3):292-302
11. [11]
  Souza R L, Lima R A, Coutinho J A P, Soares C M F, Lima Á S. Process Biochem., 2015,50(9):1459-1467
12. [12]
  Dembczyński R, Białas W, Regulski K, Jankowski T. Process Biochem., 2010,45(3):369-374
13. [13]
  Nandini K E, Rastogi N K. Appl. Biochem. Biotechnol., 2011,163(1):173-185
14. [14]
  Lü H K, Guo D, Jing Z X, Li Y H, REN B Z. Fluid Phase Equilib., 2013,341(15):23-29
15. [15]
  Mokhtarani B, Mortaheb H R, Mafi M, Amini M H. J. Chromatogr. B, 2011,879(11-12):721-726
16. [16]
  De Oliveira M C, De Abreu Filho M A N, Pessôa Filho P A. Biochem. Eng. J., 2007,37(3):311-318
17. [17]
  Berggren K, Johansson H O, Tjerneld F. J. Chromatogr. A, 1995,718(1):67-79
18. [18]
  Kalaivani S, Regupathi. J. Bioprocess Biosyst. Eng., 2013,36(10):1475-1483

扫一扫看文章

计量

PDF下载量: 0
文章访问数: 1318
HTML全文浏览量: 160

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

聚合物/磷酸盐双水相分离乳清分离蛋白中α-乳白蛋白和β-乳球蛋白条件的优化

English

Separation of α-Lactalbumin and β-Lactoglobulin in Whey Protein Isolate by Aqueous Two-phase System of Polymer/Phosphate

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

聚合物/磷酸盐双水相分离乳清分离蛋白中α-乳白蛋白和β-乳球蛋白条件的优化

English

Separation of α-Lactalbumin and β-Lactoglobulin in Whey Protein Isolate by Aqueous Two-phase System of Polymer/Phosphate

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

CCS Chemistry：颜徐州、鲍哲南： 你的皮肤可能是一片SPMs

CCS Chemistry：工作高效不疲劳，只须让催化剂动起来

CCS Chemistry：静电组装导向聚合- 聚电解质纳米凝胶量化制备新策略

CCS Chemistry：金黄色葡萄球菌醛基脱氢酶是如何识别特异性底物的？

计量

文章相关

通讯作者: 陈斌, bchen63@163.com

中国化学会秘书处

CCS Chemistry：颜徐州、鲍哲南：你的皮肤可能是一片SPMs