Cloning and analysis of a new aliphatic amidase gene from Rhodococcus erythropolis TA37

• Published in: Russian journal of genetics Vol. 50; no. 10; pp. 1009 - 1016
• Main Authors: Lavrov, K. V., Karpova, I. Yu, Epremyan, A. S., Yanenko, A. S.
• Format: Journal Article
• Language: English
• Published: Moscow Pleiades Publishing 01-10-2014
• Subjects: Animal Genetics and Genomics; Biomedical and Life Sciences; Biomedicine; Escherichia coli; Genetics of Microorganisms; Human Genetics; Microbial Genetics and Genomics; Rhodococcus; Rhodococcus erythropolis; TA37 Strain; Rhodococcus Strain; Acetamide; Acetanilide; Thiostrepton
• ISSN: 1022-7954; 1608-3369
• DOI: 10.1134/S1022795414100056

A new aliphatic amidase gene (
ami
), having a less than 77% level of similarity with the nearest homologs, was identified in the
Rhodococcus erythropolis
TA37 strain, which is able to hydrolyze a wide range of amides. The amidase gene was cloned within a 3.7 kb chromosomal locus, which also contains putative acetyl-CoA ligase and ABC-type transporter genes. The structure of this locus in the
R. erythropolis
TA37 strain differs from the structure of loci in other Rhodococcus strains. The amidase gene is expressed in
Escherichia coli
cells. It was demonstrated that amidase (generated in the recombinant strain) efficiently hydrolyzes acetamide (aliphatic amide) and does not use 4′-nitroacetanilide (N-substituted amide) as a substrate. Insertional inactivation of the amidase gene in the
R. erythropolis
TA37 strain results in a considerable decrease (by at least 6–7 times) in basal amidase activity, indicating functional amidase activity in the
R. erythropolis
TA37 strain.