Vol 37(2021) N 6 p. 25-33; DOI 10.21519/0234-2758-2021-37-6-25-33
D.O. Osipov1*, V.Yu. Matys2, V.A. Nemashkalov2, A.M. Rozhkova1, I.A. Shashkov1, A.D. Satrutdinov1, E.G. Kondratyeva1, and A.P. Sinitsyn1,3

Cloning, Isolation, and Properties of a New Recombinant Tannase from the Aspergillus niger Fungus

1Fundamentals of Biotechnology Federal Research Center, Russian Academy of Sciences, Moscow, 119071, Russia
2Skryabin Institute of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Pushchino, 142290, Russia
3Lomonosov Moscow State University, Moscow, 119991, Russia

Received - 20.05.2021; Accepted - 22.07.2021


A new recombinant Aspergillus niger tannase (tannin acyl hydrolase) produced by the Penicillium verruculosum fungus has been studied. A strain with a high level of extracellular tannase (TAN2) secretion (80% of the total extracellular protein) was obtained by cloning the tan2 gene (PDB Acc. No: MT828303) into the recipient strain. The tannase enzyme preparation degraded tannins in black tea extracts. TAN2 was isolated in homogeneous form using chromatographic methods; the enzyme had a high activity against gallotannin (53 U/mg) and less activity against propyl gallate (4.7 U/mg). Homogeneous TAN2 showed temperature and pH optima of 45 °C and 3.5, respectively. At a temperature of 50 °C, TAN2 retained above 80% activity for 3 h; at 60 °C, it retained about 75% of its activity for 90 min; at 70 °C, the enzyme was completely inactivated within 10 min. Tannase was characterized by a high tolerance to NaCl, the activity against gallotannin exceeded 50% of the initial value in solutions with a salt concentration of up to 5 M. The tannase activity was stimulated by Ca2+, Mg2+, Zn2+, Mn2+, Cu2+, Cd2+, Pb2+ by 3--64% and inhibited by 4-65% in the presence of Co2+, Fe3+ and Fe2+ ions.

tannase, tannins, Aspergillus niger, Penicillium verruculosum

This work was supported by the Ministry of Science and Higher Education of the Russian Federation.