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Vol 33(2017) N 6 p. 12-27; DOI 10.21519/0234-2758-2017-33-6-12-27
A.S. Aksambayeva1*, L.R. Zhaparova1, Zh.S. Shagyrova1, E. Zhiyenbay2, T.S. Nurgozhin2, E.M. Ramankulov2, and A.V. Shustov1

Recombinant Tyrosinase from Verrucomicrobium spinosum: Isolation, Characteristics and Use for Production of Protein with Adhesive Properties

1The National Center for Biotechnology, 010000, Astana Republic of Kazakhstan
2The Nazarbayev University, 010000, Astana Republic of Kazakhstan

*altyn-ai@bk.ru
Received - 12.05.2017; Accepted - 06.10.2017

References

The recombinant tyrosinase from the Verrucomicrobium spinosum bacterium has been obtained using the heterological expression in E. coli and isolated in the pure state to determine a number of its biochemical properties (kinetics of the enzymatic reaction, effect of an inhibitor and activator, and pH optimum). Co-expression of the genes for tyrosinase and recombinant adhesive mussel protein in one producer strain was used to introduce post-translational modifications into the adhesive protein. These modifications included the tyrosine transformation into DOPA residues which gives the protein an ability of molecular adhesion in aqueous medium. The resulting adhesive protein showed unexpectedly high adhesion strength of 2.5 MPa, which exceeds the corresponding values for natural adhesive proteins and is by a few tens times superior to the existing medical fibrin-based medical glues.

tyrosinase, enzymatic activity, DOPA, adhesive protein, adhesion strength, biocompatible glue.

The work was supported by the Grant from the Ministry of Education and Science of the Republic of Kazakhstan (0115РК01798) «Bioadhesive Microparticles Releasing Antimicrobial Peptides, a New Preparation for Prophylaxis and Therapy of Caries».



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