Cloning and expression of the L-1-amino-2-propanol dehydrogenase gene from Rhodococcus erythropolis, and its application to double chiral compound production.
The gene encoding NADP(+)-dependent L-1-amino-2-propanol dehydrogenase (AADH) of Rhodococcus erythropolis MAK154 was cloned and sequenced. A 780-bp nucleotide fragment was confirmed to be the gene encoding AADH by agreement of the N-terminal and internal amino acid sequences of the purified AADH. The gene (aadh) codes a total of 259 amino acid residues, and the deduced amino acid sequence shows similarity to several short-chain dehydrogenase/reductase family proteins. An expression vector, pKKAADH, which contains the full length aadh was constructed. Escherichia coli cells possessing pKKAADH exhibited a 10.4-fold increase in specific activity as to catalysis of the reduction of (S)-1-phenyl-2-methylaminopropan-1-one (MAK), as compared with that of R. erythropolis MAK154 induced by 1-amino-2-propanol (1 mg/ml). Coexpression of aadh with a cofactor regeneration enzyme (glucose dehydrogenase) gene was also performed, and a system for sufficient production of d-pseudoephedrine from racemic MAK was constructed.
Kataoka
M
M
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-Oiwakecho, Sakyo-ku, Kyoto, 606-8502, Japan. kataoka@kais.kyoto-u.ac.jp
Ishige
T
T
Urano
N
N
Nakamura
Y
Y
Sakuradani
E
E
Fukui
S
S
Kita
S
S
Sakamoto
K
K
Shimizu
S
S
D000218:Adamantane / Q000031:analogs & derivatives* / Q000378:metabolism; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D042843:Glucose 1-Dehydrogenase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D012240:Rhodococcus / Q000201:enzymology*
D001426:Bacterial Proteins; C011327:1-(2-phenyladamant-1-yl)-2-methylaminopropane; D000429:Alcohol Oxidoreductases; D042843:Glucose 1-Dehydrogenase; C025719:L-aminopropanol dehydrogenase; D000218:Adamantane