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Structure–function relationship in the CYP74 family: Conversion of divinyl ether synthases into allene oxide synthases by site‐directed mutagenesis
Author(s) -
Toporkova Yana Y.,
Ermilova Valeria S.,
Gorina Svetlana S.,
Mukhtarova Lucia S.,
Osipova Elena V.,
Gogolev Yuri V.,
Grechkin Alexander N.
Publication year - 2013
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/j.febslet.2013.06.030
Subject(s) - mutagenesis , mutant , chemistry , atp synthase , ether , enzyme , allene , isomerase , biosynthesis , biochemistry , site directed mutagenesis , stereochemistry , organic chemistry , gene , catalysis
Non‐classical P450s of CYP74 family control several enzymatic conversions of fatty acid hydroperoxides to bioactive oxylipins in plants, some invertebrates and bacteria. The family includes two dehydrases, namely allene oxide synthase (AOS) and divinyl ether synthase (DES), and two isomerases, hydroperoxide lyase (HPL) and epoxyalcohol synthase. To study the interconversion of different CYP74 enzymes, we prepared the mutant forms V379F and E292G of tobacco (CYP74D3) and flax (CYP74B16) divinyl ether synthases (DESs), respectively. In contrast to the wild type (WT) enzymes, both mutant forms lacked DES activity. Instead, they produced the typical AOS products, α‐ketols and (in the case of the flax DES mutant) 12‐oxo‐10,15‐phytodienoic acid. This is the first demonstration of DES into AOS conversions caused by single point mutations.