Carnitine biosynthesis in Neurospora crassa: Identification of a cDNA coding for ε-N-trimethyllysine hydroxylase and its functional expression in Saccharomyces cerevisiae
Date
2002
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Abstract
The biosynthesis of L-carnitine in eukaryotic organisms was first elucidated in the ascomycete Neurospora crassa. The first step of the pathway is catalysed by ε-N-trimethyllysine hydroxylase (TMLH), which converts ε-N-trimethyllysine into β-hydroxy-N-ε-trimethyllysine in a reaction dependent on α-ketoglutarate, Fe2+ and oxygen. Here we report on the cloning of the N. crassa TMLH cDNA and its functional expression in Saccharomyces cerevisiae. The TMLH cDNA contains an open reading frame of 1413 base pairs encoding a predicted polypeptide of 471 amino acids. The Michaelis-Menten constants of the heterologously expressed enzyme were determined for ε-N-trimethyllysine, α-ketoglutarate, Fe2+ and correspond to 0.33 mM, 133 μM and 46 μM, respectively. © 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
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Keywords
2 oxoglutaric acid, 6 n trimethyllysine hydroxylase, carnitine, complementary DNA, iron, oxygenase, unclassified drug, article, base pairing, biosynthesis, controlled study, molecular cloning, Neurospora crassa, nonhuman, nucleotide sequence, open reading frame, priority journal, protein expression, Saccharomyces cerevisiae, Amino Acid Sequence, Carnitine, Cloning, Molecular, Consensus Sequence, DNA, Complementary, Kinetics, Mixed Function Oxygenases, Molecular Sequence Data, Neurospora crassa, Saccharomyces cerevisiae, Sequence Alignment, Sequence Homology, Amino Acid, Eukaryota, Neurospora, Neurospora crassa, Saccharomyces, Saccharomyces cerevisiae
Citation
FEMS Microbiology Letters
210
1
210
1