Problems and prospects of using Sephacryl S-200 for separation of cellulase and xylanase complexes of Coprinus cinereus

L. Kamariah and J. S. Knapp

Abstract

The crude culture filtrates obtained from the growth of Coprinus cinereus on Solka Floc which contained cellulase and xylanase complexes, were concentrated through ultrafiltration membrane (Diaflo PM 10, molecular weight cut-off of 10 000) and later fractionated through gel filtration chromatography (Sephacryl S-200). Results from gel filtration chromatography showed that the apparent molecular weight of protein from each activity peak was very much less than expected. It appeared that the gel did not act solely as a passive molecular sieve suggesting that an adsorption or electrostatic interaction might have occurred between the enzyme and the matrix of the column. Molecular weight determination from SDS-polyacrylamide gel electrophoresis (SDS-PAGE) gave a higher and more realistic molecular weight thus supporting the idea.

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