Open AccessBiosynthesis of terpenes: Studies on 1-hydroxy-2-methyl-2-( E )-butenyl 4-diphosphate reductase
Author(s) -
Petra Adam,
Stefan Hecht,
Wolfgang Eisenreich,
J. Kaiser,
Tobias Gräwert,
D. Arigoni,
Adelbert Bacher,
Felix Rohdich
Publication year - 2002
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.182412599
Subject(s) - divalent , biochemistry , escherichia coli , chemistry , biosynthesis , enzyme , stereochemistry , reductase , gene , organic chemistry
Earlier in vivo studies showed the involvement of IspH protein in the conversion of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate into isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). We have demonstrated now that cell extract of an Escherichia coli strain engineered for hyperexpression of the ispH (lytB) gene catalyzes the in vitro conversion of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate into IPP and DMAPP. The reaction requires NADH, FAD, divalent cations (preferably Co2+), and probably one or more as-yet-unidentified proteins. The low intrinsic catalytic activities of wild-type E. coli cell extract and isolated chromoplasts of red pepper (Capsicum annuum) are enhanced by the addition of purified recombinant IspH protein.