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Engineering of the H 2 O 2 ‐binding pocket region of a recombinant manganese peroxidase to be resistant to H 2 O 2
Author(s) -
Miyazaki Chie,
Takahashi Haruo
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)03127-1
Subject(s) - phanerochaete , peroxidase , manganese , hydrogen peroxide , manganese peroxidase , chemistry , enzyme , stereochemistry , biochemistry , organic chemistry
The manganese peroxidase produced by Phanerochaete chrysosporium , which catalyzes the oxidation of Mn 2+ to Mn 3+ , is easily inactivated by the hydrogen peroxide (H 2 O 2 ) presented in the reaction. We attempted to increase H 2 O 2 resistance by the conformational stabilization around the H 2 O 2 ‐binding pocket. Based on its structural model, engineering of oxidizable Met273 located near the pocket to a non‐oxidizable Leu showed a great improvement. Furthermore, after treatment at 1 mM H 2 O 2 where the wild‐type is completely inactivated, full activity can be retained by engineering the Asn81, which might have conformational changes due to the environment of the pocket, to a non‐bulky and non‐oxidizable Ser.