Advertisement

Applied Biochemistry and Biotechnology

, Volume 78, Issue 1–3, pp 501–510 | Cite as

Partitioning invertase between a dilute water solution and generated droplets

  • Samuel Ko
  • Veara Loha
  • Liping Du
  • Ales Prokop
  • Robert D. TannerEmail author
Article

Abstract

Water droplets or mist occur naturally in the air at seashores. These water droplets carry inorganic and organic substances from the sea to the land via the air, creating fertile land in sandy coastal areas (1). The same phenomenon occurs in an air-fluidized bed bioreactor (2). In an air-fluidized bed reactor, proteins can be transferred from the bioreactor semisolid bulk phase to an enriched droplet phase. This protein transfer process (droplet fractionation) can be experimentally simulated by shaking a separatory funnel containing a dilute solution of a given protein, which can be an enzyme like invertase. The created droplets become richer in invertase (protein) than that of the original dilute solution. The droplets can then be coalesced by tranpping them and recovering the concentrated protein in the new liquid phase. Typically, in such a droplet fractionation process a collected enzyme can be degraded in its ability tocatalyze a chemical reaction. In this article, we explore whether the initial solution pH control variable can be adjusted to minimize the decrease of enzyme activity in this process. The protein droplet recovery problem is one in which the recovered amount of desired protein (enzyme) in the droplet is maximized, subject to the minimization of the enzyme activity loss. The partition coefficient, which is the ratio between the protein concentration in the droplets and the residual solution, is maximized at approx 4.8 and occurs at pH 3.0. Here, the partition coefficient for invertase decreases as the initial solution pH increases, between pH 3.0 and 8.0. Interestingly, the initial solution surface tension seems to be inversely proportional to the partition coefficient. The partition coefficien treachesa maximum value at a surface tension value of approx 63 mN/m at pH 3.0. The enzymatic activity of the initial, the residual, and the droplet solutions all decrease as the bulksolution pH increases. Adecrease of enzymatic activity was observed in the residual bulk solution when compared with that in the initial bulk solution at all pH levels. Also, up to 90% of the invertase activity was lost in the droplets when compared to the initial bulk solution.

Index Entries

Droplet fractionation droplets invertase protein separation decrease in enzyme activity 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Blanchard, D. C. (1972), Saturday Rev. Jan. 1, 60–63.Google Scholar
  2. 2.
    Kokitkar, P. B., Hong, K., and Tanner R. D. (1990), J. Biotech. 15, 305–322.CrossRefGoogle Scholar
  3. 3.
    Wiseman, A. (1979), Topics in Enzyme and Fermentation Biotechnology 3, Ellis Horwood, New York, pp. 267–282.Google Scholar
  4. 4.
    Lampen, J. O. (1971), The Enzymes, vol. 5, Boyer, P. D., ed., Academic, New York, pp. 291–305.Google Scholar
  5. 5.
    Hong, K., Tanner, R. D., Malaney, G. W., and Danzo, B. J. (1989), Bioprocess Eng. 4, 209–215.CrossRefGoogle Scholar
  6. 6.
    Kokitkar, P. B. and Tanner, R. D. (1991), Appl. Biochem. Biotechnol. 28/29, 647–653.Google Scholar
  7. 7.
    Smith, A. E., Achremowicz, B., Karkare, M. V., and Tanner, R. D. (1992), J. Microb. Biotech. 7, 9–17.Google Scholar
  8. 8.
    Adamson, A. W. (1967), Physical Chemistry of Surfaces, 2nd. ed., Wiley, New York, pp. 73–78.Google Scholar
  9. 9.
    Bradford, M. M. (1976), Anal. Biochem. 72, 248–264.CrossRefGoogle Scholar
  10. 10.
    Miller, G. L. (1959), Anal. Chem. 31, 426–428.CrossRefGoogle Scholar
  11. 11.
    Loha, V., Nun, S. N., Sarkawi, S. S., Prokop, A., Tanner, R. D., and Vitolo, M. (1998), Revista de Farmácia e Bioquímica da Universidade de São Paulo, in press.Google Scholar
  12. 12.
    Loha, V., Prokop, A., Du, L., and Tanner, R. D. (1999), Appl. Biochem. Biotechnol. 77–79, 701–712.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 1999

Authors and Affiliations

  • Samuel Ko
    • 1
  • Veara Loha
    • 1
  • Liping Du
    • 1
  • Ales Prokop
    • 1
  • Robert D. Tanner
    • 1
    Email author
  1. 1.Chemical Engineering DepartmentVanderbilt UniversityNashville

Personalised recommendations