Open AccessPurification and Characterization of a Rhamnogalacturonase with Protopectinase Activity from Trametes sanguinea
Author(s) -
Sakamoto Mitsuhiro,
Shirane Yuichi,
Naribayashi Ikuya,
Kimura Katsumi,
Morishita Naoichi,
Sakamoto Tatsuji,
Sakai Takuo
Publication year - 1994
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1994.tb20052.x
Subject(s) - isoelectric point , size exclusion chromatography , chemistry , molecular mass , pectin , chromatography , enzyme , ion chromatography , ion exchange , biochemistry , organic chemistry , ion
In a culture filtrate of Trametes sanguinea IFO 6490, we found a protopectin‐solubilizing enzyme, protopectinase‐T, that did not degrade polygalacturonic acid. The enzyme was purified to homogeneity with hydrophobia, cation‐exchange, anion‐exchange, and size‐exclusion chromatographies. It had an apparent molecular mass of 55 kDa by SDS/PAGE and 39 kDa by size‐exclusion chromatography on Superose 12. The isoelectric point was at pH 8.1. Protopectinase‐T was stable from pH 3.0 to 6.0 and at temperatures up to 50°C. The optimum pH for enzyme activity was 4.0 at 37°C, and the optimum temperature was 50°C at pH 5.0. Protopectinase‐T catalyzed the release of highly polymerized pectin from lemon peel protopectin.