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Cloning and expression of the L-1-amino-2-propanol dehydrogenase gene from Rhodococcus erythropolis, and its application to double chiral compound production.
- Source :
-
Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2008 Sep; Vol. 80 (4), pp. 597-604. Date of Electronic Publication: 2008 Jun 27. - Publication Year :
- 2008
-
Abstract
- 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.
- Subjects :
- Adamantane metabolism
Alcohol Oxidoreductases metabolism
Bacterial Proteins metabolism
Escherichia coli genetics
Escherichia coli metabolism
Glucose 1-Dehydrogenase genetics
Glucose 1-Dehydrogenase metabolism
Molecular Sequence Data
Adamantane analogs & derivatives
Alcohol Oxidoreductases genetics
Bacterial Proteins genetics
Cloning, Molecular
Gene Expression
Rhodococcus enzymology
Subjects
Details
- Language :
- English
- ISSN :
- 1432-0614
- Volume :
- 80
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Applied microbiology and biotechnology
- Publication Type :
- Academic Journal
- Accession number :
- 18584170
- Full Text :
- https://doi.org/10.1007/s00253-008-1563-6