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Rational re‐design of the substrate binding site of flavocytochrome P450 BM3
Author(s) -
Ost Tobias W.B,
Miles Caroline S,
Murdoch Jane,
Cheung York-Fong,
Reid Graeme A,
Chapman Stephen K,
Munro Andrew W
Publication year - 2000
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(00)02267-5
Subject(s) - butyrate , stereochemistry , bacillus megaterium , carboxylate , binding site , chemistry , active site , rational design , substrate (aquarium) , biochemistry , mutant , enzyme , lysine , amino acid , biology , bacteria , fermentation , ecology , gene , genetics
Bacillus megaterium P450 BM3 is a fatty acid hydroxylase with selectivity for long chain substrates (C 12 –C 20 ). Binding or activity with substrates of chain length 13‐fold with butyrate, while the L75T/L181K double mutant has k cat / K M increased >15‐fold with hexanoate and binding ( K d ) improved >28‐fold for butyrate. Removing the arginine 47/lysine 51 carboxylate binding motif at the mouth of the active site disfavours binding of all fatty acids, indicating its importance in the initial recognition of substrates.