acetic acid buffer ph 5 0 then the enzyme activity adsorbed to the column was eluted by washing the column with a linear concentration slope 0 50 of ethylene glycol in the equilibration buffer the eluted solution is divided into 90 fractions approximately and the enzyme activity of each fraction is measured according to the measuring method described in example 1 from fractions of eluted solution active fractions total 48 ml are collected this solution is dialyzed against 0 01m phosphate buffer ph 7 0 then applied to a column of mono q ameshampharmacia biotech which is previously made equilibrated with the same buffer the activity was eluted by washing the column with a linear gradient 0 1m of sodium chloride active fractions 10 6 mi were collected and concentrated to 0 5 ml using centricon centrifuge concentrator amicon co ltd said concentrator has an ultrafiltration membrane made of cellulose derivative at the bottom of the container when mixed solution composed of enzyme and protein is poured into the container and centrifuged by 6000 r p m protein contained in the solution is remained on the membrane while water or low molecular weight ion passes through the film and recovered as a filtrated liquid and divided by a gel filtrating chromatography method and 1 2 ml of fraction having higher activity is.