PURIFICATION AND CHARACTERIZATION OF A PYRUVATED-MANNOSE-SPECIFIC XANTHAN LYASE FROM HEAT-STABLE, SALT-TOLERANT BACTERIA

A xanthanase complex secreted by a consortium of heat-stable, salt-tolerant bacteria includes a lyase that specifically removes terminal pyruvated beta-D-mannose residues from the side chains of xanthan gum. The enzyme was purified to homogeneity from the culture broth following ion-exchange chromatography and gel permeation chromatography. It consists of a single subunit of molecular weight 33,000. The enzyme is stable to 55-degrees-C for more than 6 h in 20 mM sodium phosphate buffer (pH 5.0) containing 0.25 M NaCI. Optimal enzyme activity was observed at 0.05 M NaCI and a pH of 5. The enzyme has a pI of 3.7. It does not remove unsubstituted terminal beta-D-mannose residues from xanthan side chains nor does it hydrolyze p-nitrophenyl-beta-D-mannose. Treatment of xanthan with purified lyase results in a polysaccharide containing side chains terminating in an unsaturated 4,5-ene-glucuronic acid.