Some simple methods for the purification of pectic enzymes from Aspergillus usamii

Publisher Website
Google Scholar
Abstract
In the course of an investigation of the enzymatic degradation of pectic substances by Aspergillus usamii Sakaguchi, we found that the fungus produced extracellular pectic enzymes in significant amounts, and the enzymes were able to macerate various kinds of pectic substances.22 The present paper reports and discusses the results obtained with several analytical techniques in the purification of a crude mixture of extracellular pectic enzymes produced by Aspergillus usamii Sakaguchi. Comparing chromatography on DEAE‐cellulose with DEAE‐Sephadex A‐50, the first gave optimal enzyme recovery and good separation, whereas the second demonstrated good active protein resolution but low recovery (with the exception of the 93% recovery of pectintranseliminase (PTE)). 2‐Ethoxy‐6, 9‐diamineacridine lactate (rivanol) was capable of separating the enzymes with good PTE and polymethylgalacturonase (PMG) recoveries. Acrylamide gel electrophoresis on an analytical scale separated the crude mixture into seven protein bands: the first and seventh were identified as pectinesterase (PE) and polygalacturonase (PG) respectively. The most important result of preparative electrophoresis was the isolation of PE. The question of separating PG from PMG is discussed.