Pseudo-affinity chromatography of rumen microbial cellulase on Sepharose- Cibacron Blue F3GA
Pseudo affinity adsorption of bioproducts on Sepharose-cibacron blue F3-GA was subjected to rumen microbial enzyme evaluation through batch binding and column chromatography of cellulase. The results showed that homogenizing method had better performance in the release of enzyme, so that the amount of enzyme in rumen liquor approximately doubled. Among the preconcentration methods, it was shown that freeze drying and precipitation of enzyme using ammonium sulphate were the best. Based on equilibrium adsorption data, the best temperature, contact time and elution agent were 30°C, 15 min and 0.5 M NaCl (pH 6.5), respectively. Microbial cellulase was purified from rumen liquor to 1.2 and 14.2- fold by ammonium sulphate fractionation and Sepharose-CB column chromatography, respectively. The generic sample preparation and application of the sorbent for the adsorption and separation of cellulase is discussed.
Key words: Cellulase, pseudo-affinity chromatography, purification, sepharose-cibacron blue F3-GA.