Partial Purification of Extracellular Konjakglucomannan Decomposing Enzyme from Bacillus circulans No. 215 by Affinity Chromatography

Abstract

An extracellular konjak-glucomannan decomposing enzyme produced by Bacillus circulans No. 215 was present in broth together with sugars derived from konjak. To purify the enzyme, the ordinary methods, e.g. gelfiltration and ion-exchange chromatography did not give good results. Therefore, we perfomed an affinity chromatography, using a column containing a specific adsorvent, konjakgel. The method permitted 97% exclusion of contaminating sugars from the crude enzyme fraction, and elevated the specific activity to 42.0 folds as compared with its crude extract. The partial purified enzyne degraded konjak, however it did not show the activity toward synthetic substrates, pnitrophenyl-α-Dglucopyranoside, pnitrophenyl-β-Dglucopyranoside, pnitrophenyl-α-Dmannopyranoside and pnitrophenyl-β-Dmannopyranoside.