Skip to main content
SpringerLink
Log in

Cloning, Purification, and Characterization of Tripeptidyl Peptidase from Streptomyces herbaricolor TY-21

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Tripeptidyl peptidase (TPP) is an exopeptidase that sequentially hydrolyzes tripeptides from the N-terminus of oligopeptides or polypeptides. We performed screening for isolating novel TPP-producing microorganisms from soil samples. TPP activity was observed in the culture supernatant of Streptomyces herbaricolor TY-21 by using Ala-Ala-Phe-p-nitroanilide (pNA) as the substrate. TPP from the culture supernatant was purified to approximately 790-fold. It was shown to cleave oxidized insulin B-chain, thereby with releasing tripeptide units, but not the N-terminal-protected peptide, Cbz-Ala-Ala-Phe-pNA. The TPP gene, designated tpp, was isolated from a partial genomic DNA library of S. herbaricolor TY-21. The TPP gene consisted of 1488 bp, and encoded a 133-amino acid pre-pro-peptide and a 362-amino acid mature enzyme containing conserved amino acid residues (Asp-36, His-77, and Ser-282) similar to the catalytic residues in subtilisin. TY-21 TPP belonged to the peptidase S8A family in the MEROPS database. The mature TY-21 TPP showed approximately 49% identity with tripeptidyl peptidase subtilisin-like (TPP S) from Streptomyces lividans strain 66.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Tomkinson, B., & Lindås, A. C. (2005). Tripeptidyl-peptidase II: A multi-purpose peptidase. The International Journal of Biochemistry & Cell Biology, 37, 1933–1937.

    Article  CAS  Google Scholar 

  2. Tomkinson, B. (1999). Tripeptidyl peptidases: enzymes that count. Trends in Biochemical Sciences, 24, 355–359.

    Article  CAS  Google Scholar 

  3. Doebber, T. W., Divor, A. R., & Ellis, S. (1978). Identification of a tripeptidyl aminopeptidase in the anterior pituitary gland: effect on the chemical and biological properties of rat and bovine growth hormones. Endocrinology, 103, 1794–1804.

    Article  CAS  Google Scholar 

  4. McDonald, J. K., Hoisington, A. R., & Eisenhauer, D. A. (1985). Partial purification and characterization of an ovarian tripeptidyl peptidase: a lysosomal exopeptidase that sequentially releases collagen-related (Gly-Pro-X) triplets. Biochemical and Biophysical Research Communications, 126, 63–71.

    Article  CAS  Google Scholar 

  5. Page, A. E., Fuller, K., Chambers, T. J., & Warburton, M. J. (1993). Purification and characterization of a tripeptidyl peptidase I from human osteoclastomas: evidence for its role in bone resorption. Archives of Biochemistry and Biophysics, 306, 354–359.

    Article  CAS  Google Scholar 

  6. Bålöw, R. M., Tomkinson, B., Ragnarsson, U., & Zetterqvist, O. (1986). Purification, substrate specificity, and classification of tripeptidyl peptidase II. The Journal of Biological Chemistry, 261, 2409–2417.

    PubMed  Google Scholar 

  7. Rose, C., Vargas, F., Facchinetti, P., Bourgeat, P., Bambal, R. B., Bishop, P. B., & Schwartz, J. C. (1996). Characterization and inhibition of a cholecystokinin-inactivating serine peptidase. Nature, 380, 403–409.

    Article  CAS  Google Scholar 

  8. Renn, S. C., Tomkinson, B., & Taghert, P. H. (1998). Characterization and cloning of tripeptidyl peptidase II from the fruit fly, Drosophila melanogaster. The Journal of Biological Chemistry, 273, 19173–19182.

    Article  CAS  Google Scholar 

  9. Krieger, T. J., Bartfeld, D., Jenish, D. L., & Hadary, D. (1994). Purification and characterization of a novel tripeptidyl aminopeptidase from Streptomyces lividans 66. FEBS Letters, 352, 385–388.

    Article  CAS  Google Scholar 

  10. Butler, M. J., Aphale, J. S., Binnie, C., DiZonno, M. A., Krygsman, P., Soltes, G., & Malek, L. T. (1996). Cloning and analysis of a gene from Streptomyces lividans 66 encoding a novel secreted protease exhibiting homology to subtilisin BPN. Applied Microbiology and Biotechnology, 45, 141–147.

    Article  CAS  Google Scholar 

  11. Binnie, C., Butler, M. J., Aphale, J. S., Bourgault, R., Dizonno, M. A., Krygsman, P., & Malek, L. T. (1995). Isolation and characterization of two genes encoding proteases associated with the mycelium of Streptomyces lividans 66. Journal of Bacteriology, 177, 6033–6040.

    Article  CAS  Google Scholar 

  12. Butler, M. J., Binnie, C., DiZonno, M. A., Krygsman, P., Soltes, G. A., Soostmeyer, G., & Malek, L. T. (1995). Cloning and characterization of a gene encoding a secreted tripeptidyl aminopeptidase from Streptomyces lividans 66. Applied and Environmental Microbiology, 61, 3145–3150.

    Article  CAS  Google Scholar 

  13. Reichard, U., Léchenne, B., Asif, A. R., Streit, F., Grouzmann, E., Jousson, O., & Monod, M. (2006). Sedolisins, a new class of secreted proteases from Aspergillus fumigatus with endoprotease or tripeptidyl-peptidase activity at acidic pHs. Applied and Environmental Microbiology, 72, 1739–1748.

    Article  CAS  Google Scholar 

  14. Umezawa, Y., Yokoyama, K., Kikuchi, Y., Date, M., Ito, K., Yoshimoto, T., & Matsui, H. (2004). Novel prolyl tri/tetra-peptidyl aminopeptidase from Streptomyces mobaraensis: substrate specificity and enzyme gene cloning. Journal of Biochemistry, 136, 293–300.

    Article  CAS  Google Scholar 

  15. Zotzel, J., Pasternack, R., Pelzer, C., Ziegert, D., Mainusch, M., & Fuchsbauer, H. L. (2003). Activated transglutaminase from Streptomyces mobaraensis is processed by a tripeptidyl aminopeptidase in the final step. European Journal of Biochemistry, 270, 4149–4155.

    Article  CAS  Google Scholar 

  16. Brawner, M., Poste, G., Rosenberg, M., & Westpheling, J. (1991). Streptomyces: a host for heterologous gene expression. Current Opinion in Biotechnology, 2, 674–681.

    Article  CAS  Google Scholar 

  17. Anné, J., & Van Mellaert, L. (1993). Streptomyces lividans as host for heterologous protein production. FEMS Microbiology Letters, 114, 121–128.

    Article  Google Scholar 

  18. Barrett, A. J., Rawlings, N. D., & W. J. (1998). Tripeptidyl-peptidase S. In Handbook of proteolytic enzymes, oval road (pp. 340–341). London: Academic Press.

    Google Scholar 

  19. Barrett, A. J., Rawlings, N. D., & W. J. (1998). Tripeptidyl-peptidase A, B and C. In Handbook of proteolytic enzymes, oval road (pp. 413–415). London: Academic Press.

    Google Scholar 

  20. Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–685.

    Article  CAS  Google Scholar 

  21. Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., & S. K. (1987). Current protocols in molecular biology. New York, N.Y: Greene Publishing Associates, Inc., and Wiley, Inc..

    Google Scholar 

  22. Nielsen, H., Engelbrecht, J., Brunak, S., & von Heijne, G. (1997). Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Engineering, 10, 1–6.

    Article  CAS  Google Scholar 

  23. Sleat, D. E., Donnelly, R. J., Lackland, H., Liu, C. G., Sohar, I., Pullarkat, R. K., & Lobel, P. (1997). Association of mutations in a lysosomal protein with classical late-infantile neuronal ceroid lipofuscinosis. Science, 277, 1802–1805.

    Article  CAS  Google Scholar 

  24. Vines, D. J., & Warburton, M. J. (1999). Classical late infantile neuronal ceroid lipofuscinosis fibroblasts are deficient in lysosomal tripeptidyl peptidase I. FEBS Letters, 443, 131–135.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported in part by a grant from the Japan Foundation for Applied Enzymology (to K.E.).

Author information

Authors and Affiliations

  1. Department of Applied Microbial Technology, Sojo University, Ikeda 4-22-1, Kumamoto, 860-0082, Japan

    Keisuke Ekino, Shinichi Yonei, Takuji Oka, Yoshiyuki Nomura & Takashi Shin

  2. Department of Life Science, Faculty of Science and Engineering, Setsunan University, Neyagawa, Osaka, Japan

    Hiroshi Oyama

Authors
  1. Keisuke Ekino
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shinichi Yonei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hiroshi Oyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takuji Oka
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yoshiyuki Nomura
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Takashi Shin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Keisuke Ekino.

Ethics declarations

Conflict of Interest

The authors declare no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ekino, K., Yonei, S., Oyama, H. et al. Cloning, Purification, and Characterization of Tripeptidyl Peptidase from Streptomyces herbaricolor TY-21. Appl Biochem Biotechnol 184, 239–252 (2018). https://doi.org/10.1007/s12010-017-2547-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-017-2547-8

Keywords

Search

Navigation