Genetic analysis around aminoalcohol dehydrogenase gene of Rhodococcus erythropolis MAK154: a putative GntR transcription factor in transcriptional regulation

• Published in: Applied microbiology and biotechnology Vol. 89; no. 3; pp. 739 - 746
• Main Authors: Urano, Nobuyuki, Kataoka, Michihiko, Ishige, Takeru, Kita, Shinji, Sakamoto, Keiji, Shimizu, Sakayu
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01.02.2011; Springer-Verlag; Springer; Springer Nature B.V
• Subjects: Alcohol Dehydrogenase - biosynthesis; Aldehyde dehydrogenase; Aminoalcohol dehydrogenase; Aminoalcohol metabolism; Applied Genetics and Molecular Biotechnology; Biological and medical sciences; Biomedical and Life Sciences; Biotechnology; Cloning; Dehydrogenase; Dehydrogenases; Deoxyribonucleic acid; DNA; DNA, Bacterial - chemistry; DNA, Bacterial - genetics; E coli; Enzyme kinetics; Enzymes; Fundamental and applied biological sciences. Psychology; Gene Deletion; Gene expression; Gene Expression Regulation, Bacterial; GntR family transcriptional regulator; Life Sciences; Metabolic Networks and Pathways - genetics; Metabolism; Microbial Genetics and Genomics; Microbiology; Molecular Sequence Data; Multigene Family; Open Reading Frames; Pseudoephedrine - metabolism; Repressor Proteins - genetics; Repressor Proteins - metabolism; Rhodococcus - enzymology; Rhodococcus - genetics; Rhodococcus erythropolis; Sequence Analysis, DNA; Studies; Transcription factors; Aminoalcohol metabolism; GntR family transcriptional regulator; Aminoalcohol dehydrogenase; Enzyme; Rhodococcus erythropolis; Dehydrogenase; Metabolism; Regulation(control); Gene; Bacteria; Genetics; Actinomycetes; Oxidoreductases; Transcription factor
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-010-2924-5

NADP⁺-dependent aminoalcohol dehydrogenase (AADH) of Rhodococcus erythropolis MAK154 catalyzes the reduction of (S)-1-phenyl-1-keto-2-methylaminopropane ((S)-MAK) to d-pseudoephedrine, which is used as a pharmaceutical. AADH is suggested to participate in aminoalcohol or aminoketone metabolism in this organism because it is induced by the addition of several aminoalcohols, such as 1-amino-2-propanol. Genetic analysis of around the aadh gene showed that some open reading frames (ORFs) are involved in this metabolic pathway. Four of these ORFs might form a carboxysome-like polyhedral organelle, and others are predicted to encode aminotransferase, aldehyde dehydrogenase, phosphotransferase, and regulator protein. OrfE, a homologous ORF of the FadR subfamily of GntR transcriptional regulators, lies downstream from aadh. To investigate whether or not orfE plays a role in the regulation of aadh expression, the gene disruption mutant of R. erythropolis MAK154 was constructed. The ΔorfE strain showed higher AADH activity than wild-type strain. In addition, a transformed strain, which harbored multi-orfE, showed no AADH activity even in the induced condition with 1-amino-2-propanol. These results suggest that OrfE is a negative regulator that represses aadh expression in the absence of 1-amino-2-propanol.