Cloning and expression of the l-1-amino-2-propanol dehydrogenase gene from Rhodococcus erythropolis, and its application to double chiral compound production

• Published in: Applied microbiology and biotechnology Vol. 80; no. 4; pp. 597 - 604
• Main Authors: Kataoka, M, Ishige, T, Urano, N, Nakamura, Y, Sakuradani, E, Fukui, S, Kita, S, Sakamoto, K, Shimizu, S
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01.09.2008; Springer Berlin Heidelberg; Springer; Springer Nature B.V
• Subjects: Adamantane - analogs & derivatives; Adamantane - metabolism; Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - metabolism; Amino acids; Asymmetric reduction; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biological and medical sciences; Biotechnologically Relevant Enzymes and Proteins; Biotechnology; Catalysis; Cloning; Cloning, Molecular; Dehydrogenase; Dehydrogenases; E coli; Enzymes; ephedrine; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Fundamental and applied biological sciences. Psychology; Gene Expression; Genes; Glucose; Glucose 1-Dehydrogenase - genetics; Glucose 1-Dehydrogenase - metabolism; l-1-amino-2-propanol dehydrogenase; Life Sciences; Microbial Genetics and Genomics; Microbiology; Molecular Sequence Data; Peptides; Rhodococcus - enzymology; Rhodococcus erythropolis; Studies; Asymmetric reduction; Ephedrine; 1-amino-2-propanol dehydrogenase; L-1-amino-2-propanol dehydrogenase; Enzyme; Rhodococcus erythropolis; Chiral compound; Rhodococcus erythropolis Asymmetric reduction; Dehydrogenase; Reduction; Gene; Production; Bacteria; Actinomycetes; Oxidoreductases; Application
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-008-1563-6

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.