Purification and Crystallization of a Protopectin-solubilizing Enzyme fromTrichosporon penicillatum

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Abstract
A protopectin-solubilizing enzyme (an endo-polygalacturonase) was purified and crystallized, with an overall yield of 75%, from the culture filtrate of Trichosporon penicillatum SNO-3 by a procedure involving ammonium sulfate fractionation and chromatographies on CM-Sephadex C-50 and Sephadex G-75 columns. The purified enzyme was homogeneous on polyacrylamide gel electrophoresis and ultracentrifugation. The sedimentation coefficient (s20, W) was determined to be 3.66 S, and the molecular weight was determined to be 30,000 by gel filtration and ultracentrifugation. The enzyme was a glycoprotein containing 1.7% sugar, and had an isoelectric point of around pH 7.8. The enzyme catalyzed the liberation of a pectin substance from protopectin of various plant tissues. The enzyme was different immunologically from endo-polygalacturonases produced by Saccharomyces fragilis and Aspergillus niger.