Skip to main content
SpringerLink
Log in

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

  • Genetics of Microorganisms
  • Published:
Russian Journal of Genetics Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Debabov, V.G. and Yanenko, A.S., Biocatalytic hydrolyse of nitriles, Obz. Zh. Khim., 2011, vol. 1, no. 4, pp. 376–394.

    Google Scholar 

  2. Bork, P. and Koonin, E.V., A new family of carbon-nitrogen hydrolases, Protein Sci., 1994, vol. 3, no. 8, pp. 1344–1346.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  3. Fournand, D. and Arnaud, A., Aliphatic and enantioselective amidases: from hydrolysis to acyl transfer activity, J. Appl. Microbiol., 2001, vol. 91, no. 3, pp. 381–393.

    Article  PubMed  CAS  Google Scholar 

  4. Lavrov, K.V. and Yanenko, A.S., Cloning of new acylamidase gene from Rhodococcus erythropolis and its expression in Escherichia coli, Russ. J. Genet., 2013, vol. 49, no. 10, pp. 1078–1081.

    Article  CAS  Google Scholar 

  5. Ryabchenko, L.E., Polyakova, I.N., and Yanenko, A.S., Mobilizable plasmid vectors capable of conjugative transfer from Escherichia coli to Rhodococcus and their application to construction of Rhodococcus strains, Biotekhnol., 2005, no. 5, pp. 6–13.

    Google Scholar 

  6. Birnboim, H.C. and Doly, J., A rapid alkaline extraction procedure for screening recombinant plasmid DNA, Nucleic Acids Res., 1979, vol. 7, no. 6, pp. 1513–1523.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  7. Ryabchenko, L.E., Podchernyaev, D.A., Kotlova, E.K., and Yanenko, A.S., Cloning the amidase gene from Rhodococcus rhodochrous M8 and its expression in Escherichia coli, Russ. J. Genet., 2006, vol. 42, no. 8, pp. 886–892.

    Article  CAS  Google Scholar 

  8. Xu, M., Yu, H.M., Tan, T.W., et al., Molecular cloning of an amidase gene from Nocardia sp. and its expression in Escherichia coli, Sheng Wu Gong Cheng Xue Bao, 2006, vol. 22, no. 4, pp. 682–685.

    PubMed  CAS  Google Scholar 

  9. Soubrier, F., Levy-Schil, S., Mayaux, J.F., et al., Cloning and primary structure of the wide-spectrum amidase from Brevibacterium sp. R312: high homology to the amiE product from Pseudomonas aeruginosa, Gene, 1992, vol. 116, no. 1, pp. 99–104.

    Article  PubMed  CAS  Google Scholar 

  10. Sharma, M., Sharma, N.N., and Bhalla, T.C., Purification studies on a thermo-active amidase of Geobacillus pallidus BTP-5x MTCC 9225 isolated from thermal springs of Tatapani, Appl. Biochem. Biotechnol., 2013, vol. 169, no. 1, pp. 1–14.

    Article  PubMed  CAS  Google Scholar 

  11. Solovyev, V. and Salamov, A., Automatic annotation of microbial genomes and metagenomic sequences, in Metagenomics and Its Applications in Agriculture, Biomedicine and Environmental Studies, Li, R.W., Ed., New York: Nova Science, 2011, pp. 61–78.

    Google Scholar 

  12. Bigey, F., Chebrou, H., Fournand, D., and Arnaud, A., Transcriptional analysis of the nitrile-degrading operon from Rhodococcus sp. ACV2 and high level production of recombinant amidase with an Escherichia coli-T7 expression system, J. Appl. Microbiol., 1999, vol. 86, no. 5, pp. 752–760.

    Article  PubMed  CAS  Google Scholar 

  13. Lavrov, K.V., Larikova, G.A., and Yanenko, A.S., Novel biocatalytic process of N-substituted acrylamide synthesis, Appl. Biochem. Microbiol., 2013, vol. 49, no. 8, pp. 1–4.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Research Institute for Genetics and Selection of Industrial Microorganisms, Moscow, 117545, Russia

    K. V. Lavrov, I. Yu. Karpova & A. S. Yanenko

  2. Scientific and Production Center “Armbiotechnology,”, National Academy of Sciences of Republic of Armenia, Yerevan, 0056, Republic of Armenia

    A. S. Epremyan

Authors
  1. K. V. Lavrov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. I. Yu. Karpova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. S. Epremyan
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. S. Yanenko
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to A. S. Yanenko.

Additional information

Original Russian Text © K.V. Lavrov, I.Yu. Karpova, A.S. Epremyan, A.S. Yanenko, 2014, published in Genetika, 2014, Vol. 50, No. 10, pp. 1145–1153.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Lavrov, K.V., Karpova, I.Y., Epremyan, A.S. et al. Cloning and analysis of a new aliphatic amidase gene from Rhodococcus erythropolis TA37. Russ J Genet 50, 1009–1016 (2014). https://doi.org/10.1134/S1022795414100056

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1022795414100056

Keywords

Search

Navigation