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Related Experiment Videos

Partitioning invertase between a dilute water solution and generated droplets.

S Ko1, V Loha, L Du

  • 1Chemical Engineering Department, Vanderbilt University, Nashville, TN 37235, USA.

Applied Biochemistry and Biotechnology
|August 12, 2004
PubMed
Summary

Controlling initial solution pH during protein droplet fractionation is key. Optimal pH 3.0 maximizes protein concentration in droplets while minimizing enzyme activity loss, crucial for bioreactor applications.

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Area of Science:

  • Biochemical Engineering
  • Separation Science
  • Protein Chemistry

Background:

  • Natural phenomena like sea mist concentrate substances.
  • Air-fluidized bed bioreactors mimic this, transferring proteins to a droplet phase.
  • This process, known as droplet fractionation, enriches proteins but can reduce enzyme activity.

Purpose of the Study:

  • To investigate the effect of initial solution pH on enzyme activity loss during droplet fractionation.
  • To optimize pH for maximizing protein recovery and minimizing enzyme degradation.
  • To understand the relationship between pH, surface tension, and protein partitioning.

Main Methods:

  • Simulating droplet fractionation by shaking a protein solution (invertase) in a separatory funnel.
  • Measuring protein concentration and enzymatic activity in initial, residual, and droplet phases.

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  • Varying initial solution pH and surface tension to assess their impact on the partition coefficient and activity loss.

    • Main Results:

    • The partition coefficient (droplet to residual concentration ratio) peaked at approximately 4.8 near pH 3.0.
    • Protein partitioning decreased as initial solution pH increased from 3.0 to 8.0.
    • Enzyme activity loss increased with rising bulk solution pH, with up to 90% activity lost in droplets at certain pH levels.

    Conclusions:

    • Initial solution pH is a critical variable for optimizing droplet fractionation of proteins like invertase.
    • pH 3.0 appears to be the optimal condition for maximizing protein concentration in droplets while mitigating activity loss.
    • Further research into surface tension effects and activity preservation is warranted for industrial applications.