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LytB protein catalyzes the terminal step of the 2‐ C ‐methyl‐ D ‐erythritol‐4‐phosphate pathway of isoprenoid biosynthesis
Author(s) -
Altincicek Boran,
Duin Evert C.,
Reichenberg Armin,
Hedderich Reiner,
Kollas Ann-Kristin,
Hintz Martin,
Wagner Stefanie,
Wiesner Jochen,
Beck Ewald,
Jomaa Hassan
Publication year - 2002
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(02)03726-2
Subject(s) - stereochemistry , chemistry , thermophile , escherichia coli , biochemistry , eubacterium , enzyme , aquifex aeolicus , biosynthesis , biology , bacteria , gene , genetics
Recombinant LytB protein from the thermophilic eubacterium Aquifex aeolicus produced in Escherichia coli was purified to apparent homogeneity. The purified LytB protein catalyzed the reduction of ( E )‐4‐hydroxy‐3‐methyl‐but‐2‐enyl diphosphate (HMBPP) in a defined in vitro system. The reaction products were identified as isopentenyl diphosphate and dimethylallyl diphosphate. A spectrophotometric assay was established to determine the steady‐state kinetic parameters of LytB protein. The maximal specific activity of 6.6±0.3 μmol min −1 mg −1 protein was determined at pH 7.5 and 60°C. The k cat value of the LytB protein was 3.7±0.2 s −1 and the K m value for HMBPP was 590±60 μM.