SIMULTANEOUS PRODUCTION AND PURIFICATION OF BACILLUS-SUBTILIS ALPHA-AMYLASE

Production of alpha-amylase with B. subtilis CCM 2722 in an aqueous tw o-phase polyethylene glycol/dextran System integrated with product pur ification by affinity chromatography on crosslinked starch during cult ivation was studied. The medium was drawn from the bioreactor to the e xternal settler during fermentation. After phase separation in the set tler the dextran-rich bottom phase with cells was returned to the bior eactor. The PEG-rich top phase was pumped to the column with crosslink ed starch and returned to the bioreactor after alpha-amylase adsorptio n. The same volumetric productivities, 0.53 U/mL/h, were reached in bo th batch and described process, but total productivity of the latter m ethod was much higher owing to shortening upstream and downstream proc essing time. The enzyme of 98% homogenity in 95% yield was obtained af ter its elution from the column.