Purification of a plant peroxidase using reversibly soluble ion-exchange polymer

被引:18
作者
Aruna, N [1 ]
Lali, A [1 ]
机构
[1] Univ Mumbai, Dept Chem Technol, Div Chem Engn, Matunga 400019, Mumbai, India
关键词
carboxymethyl cellulose; ion exchange; peroxidase; protein precipitation; thiophilic chromatography;
D O I
10.1016/S0032-9592(01)00226-6
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A plant peroxidase localized in the root tissues of Raphanus sativus was purified partially by precipitation using a reversibly soluble/insoluble ion-exchange polymer system of carboxymethyl (CM) cellulose, calcium and polyethylene glycol. The CM-cellulose-Ca2+ -PEG system resulted in a 'negative' purification factor of more than 5 in a single step. The enzyme was further purified and concentrated by thiophilic chromatography to homogeneity. An overall purification factor of 20 was obtained with 66% yield. The final specific enzyme activity was 460 000 U mg(-1) protein. The purified peroxidase showed pH optimum at 5 and temperature optimum at 40 degreesC. The enzyme followed the normal Michelis-Menton kinetics and gave K-m of 4.78 mM with 2,2'-Azinobis [3-ethyl-benzothiazoline-(6)-sulphonic acid] as substrate. (C) 2001 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:431 / 437
页数:7
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