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Related Experiment Videos

Rapid affinity purification processes for cyclodextrin glycosyltransferase from Bacillus circulans.

A Rosso1, S Ferrarotti, M V Miranda

  • 1Departamento de Ciencias Básicas, Universidad Nacional de Luján, Rutas 5 y 7, 6700, Luján, Buenos Aires, Argentina.

Biotechnology Letters
|September 15, 2005
PubMed
Summary

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Two methods for purifying cyclodextrin glycosyltransferase (CGTase) from Bacillus circulans were developed. Affinity precipitation and aqueous two-phase partition offer rapid, scalable purification of this important enzyme.

Area of Science:

  • Biochemistry
  • Enzyme Purification
  • Biotechnology

Background:

  • Cyclodextrin glycosyltransferase (CGTase) is an enzyme with significant industrial applications.
  • Efficient and scalable purification methods are crucial for the cost-effective production of CGTase.
  • Bacillus circulans is a common source organism for CGTase production.

Purpose of the Study:

  • To develop rapid and scalable purification methods for cyclodextrin glycosyltransferase (CGTase).
  • To compare the efficiency of affinity precipitation and aqueous two-phase partition for CGTase purification.

Main Methods:

  • Development of an affinity precipitation method using starch for CGTase adsorption and elution with alpha-cyclodextrin.
  • Optimization of affinity precipitation using factorial design.

Related Experiment Videos

  • Implementation of a two-step aqueous two-phase partition procedure for CGTase purification.

    • Main Results:

    • Affinity precipitation achieved 80% CGTase adsorption and 65% recovery with a purification factor of 17.
    • Aqueous two-phase partition yielded a 72% CGTase recovery with a higher purification factor of 37.
    • Both methods demonstrated rapid and easy scale-up potential.

    Conclusions:

    • Both affinity precipitation and aqueous two-phase partition are effective and scalable methods for CGTase purification.
    • Aqueous two-phase partition offers a higher purification factor compared to affinity precipitation.
    • These methods facilitate efficient recovery of cyclodextrin glycosyltransferase from Bacillus circulans.