Characterization of L-lysine 6-aminotransferase and its structural gene from Flavobacterium lutescens IFO3084.
L-Lysine 6-aminotransferase (LAT) is an enzyme involved in L-lysine catabolism in a wide range of living organisms. LAT from Flavobacterium lutescens IFO3084 was purified, and its structural gene (lat) was cloned, sequenced and expressed in Escherichia coli. Native PAGE analysis of purified LAT gave a single band corresponding to a molecular weight of about 110,000. lat encoded a protein of 493 amino acids with a deduced molecular weight of 53,200, which is very close to that of purified LAT determined on SDS-PAGE. Expression of lat in E. coli revealed that lat encodes a single subunit protein leading to LAT activity. These data suggested that LAT from F. lutescens IFO3084, like most other aminotransferases, is derived from a single ORF and is active as a homodimer.
Fujii
T
T
Central Research Laboratories, Mercian Corp., Johnan, Fujisawa 251-0057, Japan. tfujii@city-fujisawa.ne.jp
Narita
T
T
Agematu
H
H
Agata
N
N
Isshiki
K
K
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology; D005417:Flavobacterium / Q000201:enzymology*; D015870:Gene Expression; D005796:Genes / Q000235:genetics*; D005822:Genetic Vectors; D051306:L-Lysine 6-Transaminase; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D000637:Transaminases / Q000235:genetics* / Q000302:isolation & purification
D017931:DNA Primers; D011994:Recombinant Proteins; D000637:Transaminases; D051306:L-Lysine 6-Transaminase