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Preparation and Purification of Recombinant Dipeptidyl Peptidase 4 from Tenebrio molitor

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Abstract

Dipeptidyl peptidase 4—is a unique proline-specific peptidase, capable to hydrolyze the bonds, formed by proline amino acid residue, cleaving dipeptides from N-terminus of peptides and proteins, containing this imino acid in P1 position. Recombinant dipeptidyl peptidase 4 from the insect Tenebrio molitor was prepared for the first time in the Pichia pastoris protein expression system; a method for its purification was proposed. The authenticity of the obtained recombinant enzyme was confirmed by mass spectrometry. The use of the obtained preparation of the T. molitor enzyme is promising for the hydrolysis of resistant to proteolysis proline-rich peptides and proteins, particularly for prolamins – the main storage proteins of cereal seeds, since they are not fully hydrolyzed by human digestive enzymes and cause autoimmune gastrointestinal celiac disease in the susceptible group of people.

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ACKNOWLEDGMENTS

The authors are grateful to Dr. M.V. Serebryakova for the mass spectrometry analysis of the recombinant DPP4 preparation.

Funding

The work was financially supported by the Russian Foundation for Basic Research together with the Foundation for Support of Scientific and Project Activities of Students, Postgraduates, and Young Scientists “National Intellectual Development” (project no. 17-34-80158 mol_ev_a), the Russian Foundation for Basic Research (project nos. 17-54-61008 Egypt_a and 18-04-01221_a), and the Fund for Assistance to Small Innovative Enterprises (FASIE) (UMNIK, project no. 8874/GU/2015 (0018984).

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Authors and Affiliations

  1. Department of Chemistry, Moscow State University, 119991, Moscow, Russia

    V. F. Tereshchenkova & I. Yu. Filippova

  2. Bach Institute of Biochemistry, Fundamental Bases of Biotechnology Federal Research Center, Russian Academy of Sciences, 119071, Moscow, Russia

    E. V. Klyachko & S. V. Benevolensky

  3. Belozersky Research Institute of Physico-Chemical Biology, Moscow State University, 119991, Moscow, Russia

    M. A. Belozersky, Ya. E. Dunaevsky & E. N. Elpidina

Authors
  1. V. F. Tereshchenkova
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  2. E. V. Klyachko
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  3. S. V. Benevolensky
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  4. M. A. Belozersky
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  5. Ya. E. Dunaevsky
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  6. I. Yu. Filippova
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  7. E. N. Elpidina
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Corresponding author

Correspondence to E. N. Elpidina.

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Conflict of interests. The authors declare that they have no conflict of interest.

Statement on the welfare of animals. All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

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Translated by G. Levit

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Tereshchenkova, V.F., Klyachko, E.V., Benevolensky, S.V. et al. Preparation and Purification of Recombinant Dipeptidyl Peptidase 4 from Tenebrio molitor. Appl Biochem Microbiol 55, 218–223 (2019). https://doi.org/10.1134/S0003683819030141

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  • DOI: https://doi.org/10.1134/S0003683819030141

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