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Reverse Micellar Extraction of β-Galactosidase from Barley (Hordeum vulgare)

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Abstract

The reverse micellar system of sodium bis(2-ethylhexyl)sulfosuccinate (AOT)/isooctane was used for the extraction and primary purification of β-galactosidase (EC 3.2.1.23) from the aqueous extract of barley (Hordeum vulgare) for the first time. The process parameters such as the concentration of the surfactant, the volume of the sample injected, and its protein concentration, pH, and ionic strength of the initial aqueous phase for forward extraction, buffer pH, and salt concentration for back extraction are varied to optimize the extraction efficiency. Studies carried out with both phase transfer and injection mode of reverse micellar extraction confirmed the injection mode to be more suitable for β-galactosidase extraction. The extent of reverse micellar solubilization of proteins increased with an increase in protein concentration of the feed sample. However, back extraction efficiency remained almost constant (13–14.4%), which indicates the selectivity of AOT reverse micelles for a particular protein under given experimental conditions. β-Galactosidase was extracted with an activity recovery of 98.74% and a degree of purification of 7.2-fold.

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References

  1. Flowers, H. M., & Sharon, N. (1978). Advances in Enzymology, 48, 29–95.

    Article  Google Scholar 

  2. Simos, G., & Georgatsos, J. G. (1988). Biochimica et Biophysica Acta, 967, 17–24.

    CAS  Google Scholar 

  3. Gekas, V., & Lopez-Leiva, M. (1985). Process Biochemistry, 20, 2–12.

    CAS  Google Scholar 

  4. Agrawal, K. M. L., & Bahl, O. P. (1968). Journal of Biological Chemistry, 243(1), 103–111.

    CAS  Google Scholar 

  5. Biswas, S., Kayastha, A. M., & Seckler, R. (2003). Journal of Plant Physiology, 160, 327–337.

    Article  CAS  Google Scholar 

  6. Balasubramaniam, S., Lee, H. C., Lazan, H., Othman, R., & Ali, Z. M. (2005). Phytochemistry, 66, 153–163.

    Article  CAS  Google Scholar 

  7. Simos, G., Giannakouros, T., & Georgatsos, J. G. (1989). Phytochemistry, 28, 2587–2592.

    Article  CAS  Google Scholar 

  8. Harikrishna, S., Srinivas, N. D., Raghavarao, K. S. M. S., & Karanth, N. G. (2001). Advances in Biochemical Engineering, Biotechnology, 75, 119–183.

    Google Scholar 

  9. Hemavathi, A. B., Hebbar, U. H., & Raghavarao, K. S. M. S. (2007). Journal of Chemical Technology and Biotechnology, 82(11), 985–992.

    Article  CAS  Google Scholar 

  10. Hebbar, U. H., & Raghavarao, K. S. M. S. (2007). Process Biochemistry, 42(12), 1602–1608.

    Article  CAS  Google Scholar 

  11. Streitner, N., Carsten, Vob, & Flaschel, E. (2007). Journal of Biotechnology, 131(2), 188–196.

    Article  CAS  Google Scholar 

  12. Manocha, B., & Gaikar, V. G. (2006). Separation Science and Technology, 41, 3279–3296.

    Article  CAS  Google Scholar 

  13. Mathew, D. S., & Juang, R. S. (2005). Biochemical Engineering Journal, 25, 219–225.

    Article  CAS  Google Scholar 

  14. Dekker, M., Hilhorst, R., & Laane, C. (1989). Analytical Biochemistry, 178, 217–226.

    Article  CAS  Google Scholar 

  15. Goto, M., Kondo, K., & Nakashio, F. (1990). Journal of Chemical and Engineering Japan, 23, 513–515.

    Article  CAS  Google Scholar 

  16. Kinugasa, T., Hisamatsu, A., Watanabe, K., & Takeuchi, H. (1994). Journal of Chemical and Engineering Japan, 27, 557–562.

    Article  CAS  Google Scholar 

  17. Kuboi, R., Mori, Y., & Komasawa, I. (1990). Kagaku Kogaku Ronbunshu, 16, 1060–1066.

    CAS  Google Scholar 

  18. Shiomori, K., Kawano, Y., Kuboi, R., & Komasawa, I. (1995). Journal of Chemical and Engineering Japan, 28, 803–809.

    Article  CAS  Google Scholar 

  19. Giovenco, S., Verheggen, F., & Laane, C. (1987). Enzyme and Microbial Technology, 9, 470–473.

    Article  CAS  Google Scholar 

  20. Shiomori, K., Kawano, Y., Kuboi, R., & Komasawa, I. (2000). Journal of Chemical and Engineering Japan, 33, 800–804.

    Article  CAS  Google Scholar 

  21. Shiomori, K., Kawano, Y., Kuboi, R., & Komasawa, I. (1994). Journal of Chemical and Engineering Japan, 27, 410–414.

    Article  CAS  Google Scholar 

  22. Yamada, Y., Kuboi, R., & Komasawa, I. (1994). Solvent Extraction Research and Development, Japan, 1, 167–178.

    CAS  Google Scholar 

  23. Luisi, P. L., & Magid, L. J. (1986). CRC Critical Reviews in Biochemistry, 20, 409–474.

    Article  CAS  Google Scholar 

  24. Kon-no, K., & Kitahara, A. (1971). Journal Colloid and Interface Science, 37, 469–480.

    Article  CAS  Google Scholar 

  25. Kuboi, R., Yano, K., Tanaka, H., & Komasawa, I. (1993). Kagaku Kogaku Ronbunshu, 19, 446–454.

    CAS  Google Scholar 

  26. Laemmli, U. K. (1970). Nature, 227, 680–685.

    Article  CAS  Google Scholar 

  27. Triantafillidoa, D., & Georgatsos, J. G. (2001). Journal of Protein Chemistry, 20(7), 551–562.

    Article  Google Scholar 

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Acknowledgments

The authors wish to thank Dr. V Prakash, Director, CFTRI, for his keen interest in downstream processing. One of the authors (AB Hemavathi) gratefully acknowledges CSIR, India, for the award of Fellowship.

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Authors and Affiliations

  1. Department of Food Engineering, Central Food Technological Research Institute, Mysore, 570 020, India

    A. B. Hemavathi, H. Umesh Hebbar & K. S. M. S. Raghavarao

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  1. A. B. Hemavathi
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  2. H. Umesh Hebbar
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  3. K. S. M. S. Raghavarao
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Correspondence to K. S. M. S. Raghavarao.

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Hemavathi, A.B., Umesh Hebbar, H. & Raghavarao, K.S.M.S. Reverse Micellar Extraction of β-Galactosidase from Barley (Hordeum vulgare). Appl Biochem Biotechnol 151, 522–531 (2008). https://doi.org/10.1007/s12010-008-8228-x

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