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  • Title: Affinity thermoprecipitation of yeast alcohol dehydrogenase through metal ion-promoted binding with Eudragit-bound Cibacron blue 3GA.
    Author: Guoqiang D, Benhura MA, Kaul R, Mattiasson B.
    Journal: Biotechnol Prog; 1995; 11(2):187-93. PubMed ID: 7766101.
    Abstract:
    Metal ion-promoted binding of Saccharomyces cerevisiae alcohol dehydrogenase to Cibacron Blue 3GA was used for its isolation by affinity precipitation with Eudragit-bound dye. The yeast cells were broken and the cell debris was separated by flocculation with poly(ethylene imine). The supernatant containing the enzyme activity was mixed with Eudragit--Cibacron Blue in the presence of zinc ions. The precipitation of the affinity complex was induced by the addition of 50 mM CaCl2 and a subsequent increase in temperature to 40 degrees C. The enzyme was desorbed by treating the precipitate with iminodiacetic acid solution. The procedure resulted in about 66% recovery of enzyme activity with more than 20-fold purification. Recycling of Eudragit--dye for enzyme purification was also shown to be possible.
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