Synthesis of 1-piperideine-6-carboxylic acid produced by L-lysine-epsilon-aminotransferase from the Streptomyces clavuligerus gene expressed in Escherichia coli.
The gene (lat) encoding L-lysine epsilon-aminotransferase (LAT) in Streptomyces clavuligerus was cloned and expressed in Escherichia coli. Nucleotide sequence analysis of lat predicted a single open reading frame (ORF) of 1371 bp, encoding a polypeptide of 457 amino acids with calculated molecular mass of 49.89 kDa. S. clavuligerus LAT was grouped into aminotransferase subfamily II of alpha family on the basis of sequence homology. A model system composed of the recombinant LAT in phosphate buffer was set up to study the biosynthesis of 2-acetyltetrahydropyridine. Lysine was found to be transformed to 1-piperideine-6-carboxylic acid. 2-Acetyltetrahydropyridine was characterized from the mixture of 1-piperideine-6-carboxylic acid and methylglyoxal. For the first time, we demonstrated that the L-lysine epsilon-aminotransferase is responsible for the formation of 1-piperideine-6-carboxylic acid, which may react with methylglyoxal to generate the acylated N-heterocyclic odorant 2-acetyltetrahydropyridine.
Wu
Mei-Li
ML
Department of Food Science, National Pingtung University of Science and Technology, 912, Pingtung, Taiwan, Republic of China.
Chen
Jun-Hao
JH
Ho
Chi-Tang
CT
Huang
Tzou-Chi
TC
D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D051306:L-Lysine 6-Transaminase / Q000235:genetics* / Q000378:metabolism*; D009812:Odorants / Q000032:analysis; D010848:Picolinic Acids / Q000138:chemical synthesis*; D011725:Pyridines / Q000032:analysis / Q000378:metabolism; D011765:Pyruvaldehyde / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
C514147:2-acetyltetrahydopyridine; D010848:Picolinic Acids; D011725:Pyridines; D011994:Recombinant Proteins; C008919:delta-1-piperidine-6-carboxylic acid; D011765:Pyruvaldehyde; D051306:L-Lysine 6-Transaminase