Abstract
In this work, the forward and back extraction of soybean protein by reverse micelles was studied. The reverse micellar systems were formed by anionic surfactant sodium bis(2-ethyl hexyl) sulfosuccinate (AOT), isooctane and KCl solution. The effects of AOT concentration, aqueous pH, KCl concentration and phase volume ratio on the extraction efficiency of soybean protein were tested. Suitability of reverse micelles of AOT and Triton-X-100/AOT mixture in organic solvent toluene for soybean protein extraction was also investigated. The experimental results lead to complete forward extraction at the AOT concentration 120 mmol l−1, aqueous pH 5.5 and KCl concentration 0.8 mol l−1. The backward extraction with aqueous phase (pH 5.5) resulted in 100% extraction of soybean protein from the organic phase.




Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Melo EP, Aires-Barros MR, Cabral JMS (2001) Reverse micelles and protein biotechnology. Biotechnol Annu Rev 7:87–129
Harikrishna S, Srinivas ND, Raghavarao KSMS, Karanth NG (2002) Reverse micellar extraction for downstream processing of proteins/enzymes. Adv Biochem Eng Biotechnol 75:119–183
Hong DP, Lee SS, Kuboi R (2000) Conformational transition and mass transfer in extraction of proteins by AOT–alcohol–isooctane revere micellar systems. J Choromatogr B 743:203–213
Ruckenstein E, Nagarajan R (1980) Aggregation of amphiphiles in non-aqueous media. J Phys Chem 84:1349–1358
Regalado C, Asenjo JA, Pyle DL (1996) Studies on the purification of peroxidase from horseradish roots using reverse micelles. Enzyme Microb Technol 18:332–339
Yu YC, Chu Y, Ji JY (2003) Study of the factors affecting the forward and back extraction of yeast-lipase and its activity by reverse micelles. J Colloid Interf Sci 267:60–64
Menger FM, Yamada K (1979) Enzyme catalysis in water pools. J Am Chem Soc 1979(101):6731–6734
Douzou P, Keh E, Balny C (1979) Cryoenzymology in aqueous media: micellar solubilized water clusters. Proc Natl Acad Sci USA 76:681–684
Luisi PL, Bonner FJ, Pellegrini A, Wiget P, Wolf R (1979) Micellar solubilization of proteins in aprotic solvents and their spectroscopic characterization. Helv Chim Acta 62:740–753
Shin YO, Rodil E, Vera JH (2004) Surfactant precipitation and polar solvent recovery of a-chymotrypsin and ribonuclase-A. Biochem Eng J 17:91–97
Chou TS, Chiang BH (2004) Reversed micellar extraction of hen egg lysozyme. J Food Sci 63:399–402
Mathew DS, Juang RS (2005) Improved back extraction of papain from AOT reverse micelles using alcohols and a counter-ionic surfactant. Biochem Eng J 25:219–225
Pires MJ, Aires-Barros MR, Cabra JMS (1996) Liquid–liquid extraction of proteins with reversed micelles. Biotechnol Prog 12:290–301
Dekker M, Hilhorst R, Laane C (1989) Extraction of enzymes by reverse micelles. Chem Eng Sci 178:217–226
Hua YF, Huang YR, Qiu AY, Liu XY (2005) Properties of soy protein isolate prepared from aqueous alcohol washed soy flakes. Food Res Int 38:273–279
Chun L, Hongling W, Zhumei C, Xiaoling H, Xiansheng W, Xiaoxiong Z, Hao M (2007) Optimization of extraction and isolation for 11S and 7S globulins of soybean seed storage protein. Food Chem 102:1310–1316
Thanh VH, Shibasaki K (1976) Major proteins of soybean seeds. A straightforward fraction and their characterization. J Agric Food Chem 24:1117–1121
Nagano T, Hirotsuka M, Mori H (1992) Dynamic viscoelastic study on the gelation of 7S globulin from soybeans. J Agric Food Chem 40:941–944
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Shin Y-O, Weber ME, Vera JH (2003) Comparison of two methods to recover lysozyme from reverse micellar phases. Sep Sci Technol 38(8):1733–1748
Shiomori K, Ebuchi N, Kawano Y, Kuboi R, Kawakawa I (1998) Extraction characteristic of Bovine serum albumin using sodium bis(2-ethylhexyl) sulfosuccinate reverse micelles. J Ferment Bioeng 86:581–587
Cardoso MM, Barradas MJ, Kroner KH, Crespo JG (1999) Amino acid solubilization in cationic reversed micelles: factors affecting amino acid and water transfer. J Chem Technol Biotechnol 74:801–811
Huang SY, Lee YC (1994) Separation and purification of horseradish peroxidase from Armoracia rusticana root using reversed micellar extraction. Bioseparation 4:1–5
Harikrishna S, Srinivas ND, Raghavarao KSMS, Karanth NG (2002) Reverse micellar extraction for downstream processing of proteins/enzymes. Adv Biochem Eng Biotechnol 75:119–183
Dekker MRVK, Weijers SR, Baltussen JWA, Lanne C, Birjsterbosch BH (1986) Enzyme recovery by liquid–liquid extraction using reversed micelles. Chem Eng J 33:B27–B33
Dungan SR, Bausch TE, Hatton TA, Plucinski PK, Nitch W (1991) Interfacial transport processes in the reversed micellar extraction of proteins. J Colloid Interf Sci 145:33–50
Carlson A, Nagarajan R (1998) Release and recovery of porcine pepsin and bovine chymosin from reverse micelles: a new technique based on isopropyl alcohol addition. Biotechnol Prog 8:85–90
Shiomori K, Ebuchi N, Kawano Y, Kuboi R, Kawakawa I (1998) Extraction characteristic of Bovine serum albumin using sodium bis(2-ethylhexyl) sulfosuccinate reverse micelles. J Ferment Bioeng 86:581–587
Acknowledgments
This study was supported by the three science and technology Program of Guangdong Provincial Department of Science and Technology (2006B40101013).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhao, X., Li, Y., He, X. et al. Study of the factors affecting the extraction of soybean protein by reverse micelles. Mol Biol Rep 37, 669–675 (2010). https://doi.org/10.1007/s11033-009-9515-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11033-009-9515-5


