Premium
Isolation, purification and properties of the peroxidase from the hull of Glycine max var HH2
Author(s) -
Liu Wen,
Fang Jing,
Zhu WenMiao,
Gao PeiJi
Publication year - 1999
Publication title -
journal of the science of food and agriculture
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 142
eISSN - 1097-0010
pISSN - 0022-5142
DOI - 10.1002/(sici)1097-0010(199904)79:5<779::aid-jsfa252>3.0.co;2-9
Subject(s) - chemistry , peroxidase , chromatography , isoelectric point , sephadex , size exclusion chromatography , affinity chromatography , isoelectric focusing , enzyme , polyacrylamide gel electrophoresis , biochemistry
Abstract Soybean hull peroxidase (EC 1.11.1.7), an acidicperoxidase isolated from soybean ( Glycine max varHH2) hulls was purified to electrophoretic homogeneity by acombination of ammonium sulphate fractionation, DEAE‐SephadexA‐50 chromatography, concanavalin A‐Sepharose 4Baffinity chromatography and Bio‐Gel P‐60 gelfiltration. The specific activity of purified peroxidase was about57‐fold higher than that of crude extract. The yield was about16.4%. The molecular weight of the enzyme was estimated to be38 000 by SDS‐polyacrylamide gel electrophoresis. Theperoxidase was a glycoprotein containing about 18.7%carbohydrate, approximately one‐quarter of which was shown tobe glucosamine residues. It was found to have an isoelectric point of3.9. The enzyme was most active at pH 4.6 and 45°C, and wasstable in the pH range 2.5–11.5. The enzyme could tolerateheating for 10 min at 75°C without being inactivated, and at85°C, it took 40 min to inactivate the enzyme 50%,confirming that the peroxidase was a novel thermostable enzyme. Fe 2+ , Fe 3+ , Sn 2+ , CN − and N 3 − inhibitedenzyme activity, while Hg 2+ , Ag + , Pb 2+ , Cr 3+ , EDTA and SDS were notsignificantly inhibitory. © 1999 Society of Chemical Industry