Purification of the Candida utilis Extracellular Invertase using Affinity Chromatography

  • Ginalska, G. (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • Belcarz, A. (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • Lobarzewski, J. (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • Leonowicz, A. (Department of Biochemistry, Maria Curie-Sklodowska University) ;
  • Cho, Nam-Seok (School of Forest Resources, Chungbuk National University)
  • Received : 2002.01.21
  • Accepted : 2002.06.11
  • Published : 2002.09.26

Abstract

The extracellular invertase (EC 3.2.1.26) (Candida utilis) preparation was obtained from the liquid medium after desalting and freeze drying. This prepared enzyme was used for the comparative purification on 4 activated matrices by liquid column affinity chromatography method. In this method there were used controlled porous glass (CPG) silanized covalently activated by keratin, silanized silica gel and silica gel covalently covered by keratin. It was found that the invertase purification process was better using both CPG matrices (silanized CPG and keratin activated CPG) than these with two silica gel supports. Also the elution coefficient of the invertase from the two CPG columns was about 93 to 94%. Two silica gel supports found to be superior in terms of purification efficiency. The invertase purification process was confirmed by PAGE electrophoresis.

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Acknowledgement

This research was financially sponsored by the Polish Scientific Commitee Nr. 6PO4B 015 15 and Korea Agricultural R & D Promotion Center Grant (High-Value Added New Products from Waste Wood Materials, 1997-2000).