EXPRESSION OF FUNGAL MN PEROXIDASE IN ESCHERICHIA-COLI AND REFOLDING TO YIELD ACTIVE ENZYME

被引：45

作者：

WHITWAM, RE

GAZARIAN, IG

TIEN, M

机构：

[1] PENN STATE UNIV,DEPT BIOCHEM & MOLEC BIOL,UNIVERSITY PK,PA 16802

[2] PENN STATE UNIV,CTR BIOMOLEC STRUCT & FUNCT,UNIVERSITY PK,PA 16802

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1995年 / 216卷 / 03期

关键词：

D O I：

10.1006/bbrc.1995.2721

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：

The cDNA encoding Mn peroxidase isozyme H4 from Phanerochaete chrysosporium was expressed in Escherichia coli. The portion of the cDNA encoding the enzyme's signal peptide, not found in the processed holoenzyme, was deleted from the cDNA. The polypeptide was produced as inactive inclusion bodies that could be solubilized in 8 M urea and the reducing agent dithiothreitol. Reconstitution of activity was accomplished by diluting the urea concentration to 2M the presence of hemin, calcium, and oxidized glutathione. All of the additives were required for recovery of activity. The activity of the recombinant enzyme was dependent on both Mn2+ and H2O2. (C) 1995 Academic Press, Inc.

引用

页码：1013 / 1017

页数：5

共 16 条

[1]

FARRELL RL, 1987, Patent No. 878105162

[2] MN(II) OXIDATION IS THE PRINCIPAL FUNCTION OF THE EXTRACELLULAR MN-PEROXIDASE FROM PHANEROCHAETE-CHRYSOSPORIUM [J].