Effect of single‐point mutations Phe41→ His and Phe143→ Glu on folding and catalytic properties of recombinant horseradish peroxidase expressed in E. coli

FEBS Letters

Doseeva, Victoria V.¹  Galkin, Andrey G.¹  Tishkov, Vladimir I.¹  Gazaryan, Irina G.¹ 

[1] Department of Chemical Enzymology, Faculty of Chemistry, M.V. Lomonosov Moscow State University, 000958 Moscow, Russian Federation

关键词: Recombinant horseradish peroxidase;    Inclusion body;    Refolding;    Heme entrapment;    Site-specific mutagenesis;   

DOI  :  10.1016/0014-5793(94)01125-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Wild-type recombinant and Phe⁴¹→ His and Phe¹⁴³→ Glu mutant forms of horseradish peroxidase have been expressed in E. coli and reactivated from inclusion bodies with a yield of about 25%. The purified homogeneous preparations have been studied in the reaction of ABTS oxidation. The effect of mutations on heme entrapment and kinetics of ABTS oxidation demonstrates the essential role of the replaced residues in providing the hydrophobic crevice for the non-covalent heme binding.

【 授权许可】

Unknown