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Related Experiment Videos

Enzyme purification using temperature-induced phase formation.

P A Harris1, G Karlström, F Tjerneld

  • 1Department of Biochemistry, Lund University, Sweden.

Bioseparation
|January 1, 1991
PubMed
Summary
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A novel aqueous two-phase system using UCON 50-HB-5100 polymer effectively purifies enzymes from yeast. This system allows for enzyme recovery in buffer and UCON polymer recycling, enhancing purification efficiency.

Area of Science:

  • Biochemistry
  • Polymer Science
  • Separation Science

Background:

  • Aqueous two-phase systems (ATPS) are widely used for biomolecule purification.
  • Ethylene oxide/propylene oxide co-polymers offer tunable properties for ATPS development.
  • Efficient recovery and recycling of ATPS components are crucial for cost-effective bioprocessing.

Purpose of the Study:

  • To characterize a new ATPS utilizing UCON 50-HB-5100 for enzyme purification.
  • To evaluate the purification of 3-phosphoglycerate kinase and hexokinase from bakers' yeast using this ATPS.
  • To investigate the phase behavior and recyclability of the UCON 50-HB-5100 based ATPS.

Main Methods:

  • Characterization of UCON 50-HB-5100/dextran and UCON 50-HB-5100/hydroxypropyl starch phase diagrams.

Related Experiment Videos

  • Enzyme partitioning of yeast homogenate in the developed ATPS.
  • Temperature-induced phase separation for enzyme recovery and polymer recycling.

    • Main Results:

    • The UCON 50-HB-5100 polymer exhibits a cloud point of 55°C, reducible to 40°C with 0.2 M sodium sulfate.
    • Successful purification of 3-phosphoglycerate kinase and hexokinase from bakers' yeast was achieved.
    • The UCON polymer was recovered and recycled effectively after temperature-induced phase separation.

    Conclusions:

    • The UCON 50-HB-5100 based ATPS provides an efficient method for enzyme purification from yeast.
    • The tunable cloud point and recyclability of UCON 50-HB-5100 enhance the economic viability of the purification process.
    • This novel ATPS demonstrates potential for industrial-scale bioseparation applications.