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Extraction of Bovine Serum Albumin Using Reverse Micelles Formed by Hexadecyl Trimethyl Ammonium Chloride

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Abstract

The extraction of bovine serum albumin (BSA) has been investigated using reverse micelles of hexadecyl trimethyl ammonium chloride/n-octanol/isooctane. Forward extraction process parameters such as the surfactant concentration, co-solvent concentration, pH, ionic strength, and species of the initial aqueous phase were important factors affecting the extraction performance. These parameters were varied to optimize the extraction efficiency. Under the optimized conditions, forward extraction efficiencies of BSA can reach practically 99.55%. The thermodynamic study revealed that the extraction of BSA is controlled by entropy changes. Maximum back-extraction efficiency of 85.16% can be obtained at low pH values and high salt concentrations. The structures of BSA during reverse micelle extraction did not change by comparing the circular dichroism spectra of BSA back-extracted to the aqueous phase with that of feed BSA.

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References

  1. Luisi, P. L. (1985). Enzymes hosted in reverse micelles in hydrocarbon solution. Angewandte Chemie International Edition in English, 24, 439–450.

    Article  Google Scholar 

  2. Goklen, K. E., & Hatton, T. A. (1987). Liquid–liquid extraction of low molecular weight proteins by selective solubilization in reverse micelles. Separation Science and Technology, 22, 831–842.

    Article  CAS  Google Scholar 

  3. Jolivalt, C., Miner, M., & Renon, H. (1990). Extraction of α-chymotrypsin using reversed micelles. Journal of Colloid and Interface Science, 135, 85–96.

    Article  CAS  Google Scholar 

  4. Marcozz, G., Correa, N., Luisi, P. L., & Caselli, M. (1991). Protein extraction by reverse micelles: a study of the factors affecting the forward and backward transfer of α-chymotrypsin and its activity. Biotechnology and Bioengineering, 38, 1239–1246.

    Article  Google Scholar 

  5. Hebbar, H. U., Sumana, B., & Raghavarao, K. S. M. S. (2008). Use of reverse micellar systems for the extraction and purification of bromelain from pineapple wastes. Bioresource Technology, 99, 4896–4902.

    Article  Google Scholar 

  6. Dungan, S. R., Bausch, T. E., Hatton, T. A., Plucinski, P. K., & Nitch, W. (1991). Interfacial transport process in the reverse micellar extraction of proteins. Journal of Colloid and Interface Science, 145, 30–45.

    Article  Google Scholar 

  7. Kinuguasa, T., Tanahshi, S., & Takeuchi, H. (1991). Extraction of lysozyme using reversed micellar solution: distribution equilibrium and extraction rates. Industrial and Engineering Chemistry Research, 30, 2470–2476.

    Article  Google Scholar 

  8. Rho, S. G., & Kang, C. H. (2003). The factors affecting the backward-transfer of bovine serum albumin (BSA) from sodium bis (2-ethylhexyl) sulfosuccinate reverse micellar solutions. Korean Journal of Chemical Engineering, 20, 519–521.

    Article  Google Scholar 

  9. Kadam, K. L. (1986). Reverse micelles as a bioseparation tool. Enzyme and Microbial Technology, 8, 266–273.

    Article  CAS  Google Scholar 

  10. Wolbert, R. B. G., Hilhorst, R., Voskuilen, G., Nachtegaal, H., Dekker, M. R. V. K., & Bijsterbosch, B. H. (1989). Protein transfer from an aqueous phase into reversed micelles. European Journal of Biochemistry, 184, 627–633.

    Article  CAS  Google Scholar 

  11. Lu, Q., Chen, H. Y., Li, K. H., & Shi, Y. J. (1998). Transport between an aqueous phase and a CTAB/hexanol-octane reversed micellar phase. Biochemical Engineering Journal, 45, 45–52.

    Article  Google Scholar 

  12. Ayala, G. A., Kamat, S., Beckman, E. J., & Rassell, A. J. (1992). Protein extraction and activity in reverse micelles of a nonionic detergent. Biotechnology and Bioengineering, 39, 806–814.

    Article  CAS  Google Scholar 

  13. Hebbar, H. U., & Raghavarao, K. S. M. S. (2007). Extraction of bovine serum albumin using nanoparticulate reverse micelles. Process Biochemistry, 42, 1602–1608.

    Article  CAS  Google Scholar 

  14. Sun, Y., Ichikawa, S., Sugiura, S., & Furusaki, S. (1998). Affinity extraction of proteins with a reversed micellar system composed of unbound cibacron bluemodified lecithin. Biotechnology and Bioengineering, 58, 58–64.

    Article  CAS  Google Scholar 

  15. Zhang, W., Liu, H., & Chen, J. (2002). Forward and backward extraction of BSA using mixed reverse micellar system of CTAB and alkyl halides. Biochemical Engineering Journal, 12, 1–5.

    Article  Google Scholar 

  16. Zhang, T., Liu, H., & Chen, J. (1999). Affinity extraction of BSAwith reversed micellar system composed of unbound cibacron blue. Biotechnology Progress, 15, 1078–1082.

    Article  CAS  Google Scholar 

  17. Hong, D. P., Lee, S. S., & Kuboi, R. (2000). Conformational transition and mass transfer inextraction of proteins by AOT–alcohol–isooctane revere micellar systems. Journal of Chromatography B, 743, 203–213.

    Article  CAS  Google Scholar 

  18. Bradford, M. M. (1972). A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–254.

    Article  Google Scholar 

  19. Lazarova, Z., & Tonova, K. (1999). Integrated reversed micellar extraction and stripping of α-amylase. Biotechnology and Bioengineering, 63, 583–592.

    Article  CAS  Google Scholar 

  20. Krei, G. A., & Hustedt, H. (1992). Extraction of enzymes by reverse micelles. Chemical Engineering Science, 47, 90–111.

    Article  Google Scholar 

  21. Pessoa, A., Jr., & Vitolo, M. (1998). Recovery of insulinase using BDBAC reversed micelles. Process Biochemistry, 33, 291–297.

    Article  CAS  Google Scholar 

  22. Regalado, C., Asenjo, J. A., & Pyle, D. L. (1996). Studies on the purification of peroxidase from horseradish roots using reverse micelles. Enzyme and Microbial Technology, 18, 332–339.

    Article  CAS  Google Scholar 

  23. Gaikar, V. G., & Kulkarni, M. S. (2001). Selective reverse micellar extraction of penicillin acylase from E coli. Journal of Chemical Technology and Biotechnology, 76, 729–736.

    Article  CAS  Google Scholar 

  24. Harikrishna, S., Srinivas, N. D., Raghavarao, K. S. M. S., & Karanth, N. G. (2002). Reverse micellar extraction for downstream processing of proteins/enzymes. Advances in Biochemical Engineering/Biotechnology, 75, 119–183.

    Article  Google Scholar 

  25. Dungan, S. R., Bausch, T., Hatton, T. A., Plucinski, P., & Nitsch, W. (1991). Interfacial transport processes in the reversed micellar extraction of proteins. Journal of Colloid and Interface Science, 145, 30–50.

    Article  Google Scholar 

  26. Spelzini, D., Farruggia, B., & Pico, G. (2005). Features of the acid protease partition in aqueous two-phase systems of polyethylene glycol–phosphate: chymosin and pepsin. Journal of Chromatography B, 821, 60–66.

    Article  CAS  Google Scholar 

  27. Bianco-Peled, H., & Gryc, S. (2004). Binding of amino acid to “smart” sorbents:where does hydrophobicity come into play. Langmuir, 20, 169–174.

    Article  CAS  Google Scholar 

  28. Dekker, M., Hilhorst, R., & Laane, C. (1989). Isolating enzyme by reversed micelles. Analytical Biochemistry, 178, 217–226.

    Article  CAS  Google Scholar 

  29. Ono, T., Goto, M., Nakashio, F., & Hatton, T. A. (1996). Extraction behavior of hemoglobin using reversed micelles by dioleyl phosphoric acid. Biotechnology Progress, 12, 793–800.

    Article  CAS  Google Scholar 

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Acknowledgments

The authors acknowledge the financial support for this work from The National Natural Science Foundation of China (no. 20876089), the Key Technologies R&D Programme of China (no. 2007BAD87B05), and the Natural Science Foundation of Shandong Province (no. Y2007B05).

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Authors and Affiliations

  1. Key Laboratory for Special Functional Aggregated Materials of Education Ministry, School of Chemistry and Chemical Engineering, Shandong University, Jinan, 250100, People’s Republic of China

    Qingchi Sun, Yanzhao Yang, Yanmin Lu & Wenjuan Lu

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  1. Qingchi Sun
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  2. Yanzhao Yang
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  3. Yanmin Lu
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  4. Wenjuan Lu
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Correspondence to Yanzhao Yang.

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Sun, Q., Yang, Y., Lu, Y. et al. Extraction of Bovine Serum Albumin Using Reverse Micelles Formed by Hexadecyl Trimethyl Ammonium Chloride. Appl Biochem Biotechnol 163, 744–755 (2011). https://doi.org/10.1007/s12010-010-9079-9

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  • DOI: https://doi.org/10.1007/s12010-010-9079-9

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