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Kinetics of barley FA hydroperoxide lyase are modulated by salts and detergents
Author(s) -
Koeduka Takao,
Stumpe Michael,
Kajiwara Tadahiko,
Feussner Ivo
Publication year - 2003
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-003-1175-9
Subject(s) - recombinant dna , biochemistry , chemistry , enzyme , substrate (aquarium) , linoleic acid , lyase , complementary dna , escherichia coli , biology , fatty acid , gene , ecology
The cDNA from barley coding FA hydroperoxide lyase (HPL) was cloned. A recombinant protein derived from the cDNA was expressed in Escherichia coli as an active enzyme. Thus far, there have been no reports on HPL in monocotyledonous plants. The recombinant protein was shown to be most active to linolenic acid 13‐hydroperoxide, followed by linoleic acid 13‐hydroperoxide. 9‐Hydroperoxides of the FA could not be substrates for the recombinant HPL. The activity was dramatically enhanced in the presence of a detergent and/or a salt in the reaction mixture. At the same time, the kinetics of the reaction, including inactivation and the V max value of the HPL, were also greatly modulated, depending on the concentration of a monovalent cation and/or a detergent in the reaction mixture. These results suggest that these effectors induced a conformational change in barley HPL, resulting in an improvement in substrate binding and in enzyme activity.