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Studies on lipase-affinity adsorption using response-surface analysis.

E S Kamimura1, O Medieta, M I Rodrigues

  • 1Department of Chemical Process/School of Chemical Engineering/UNICAMP, CP: 6066, CEP: 13083-970, Campinas-SP-Brazil.

Biotechnology and Applied Biochemistry
|June 8, 2001
PubMed
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Optimizing lipase purification using affinity chromatography is crucial for producing pure pharmaceuticals and food additives. Lowering the bed height/column diameter ratio and superficial velocity enhances column efficiency, with maximum efficiency observed at specific L/D values.

Area of Science:

  • Biotechnology
  • Enzyme Technology
  • Chromatography

Background:

  • Lipases, enzymes from various sources, are valuable for synthesizing optically pure compounds like pharmaceuticals and food additives.
  • Affinity chromatography utilizes specific biological interactions for powerful purification.
  • Response-surface analysis offers a comprehensive method to study variable interactions in column efficiency.

Purpose of the Study:

  • To investigate the impact of bed height/column diameter ratio (L/D) and superficial velocity (V) on column efficiency during lipase purification.
  • To determine optimal parameters for maximizing column efficiency in lipase adsorption assays.

Main Methods:

  • Lipase was produced from Geotrichum sp. culture and purified using affinity chromatography in a fixed-bed column.

Related Experiment Videos

  • Response-surface analysis was employed to study the influence of L/D (2-10) and V (1-2 cm/min) on column efficiency.
  • Breakthrough curves were analyzed to assess column performance.

    • Main Results:

    • Column efficiency was inversely related to both L/D and superficial velocity, with maximum efficiency reaching 65.43%.
    • Response surface analysis indicated a minimum efficiency at an L/D ratio of approximately 8.
    • Optimal conditions for lipase purification were identified through experimental design.

    Conclusions:

    • Lowering L/D and V generally increases column efficiency for lipase purification.
    • The study provides insights into optimizing affinity chromatography parameters for enzyme purification.
    • Purified lipase from Geotrichum sp. shows potential for esterification reactions and bioaroma production.