TY - JOUR

T1 - Production of Maral (Cervus elaphus sibiricus Severtzov) Recombinant Chymosin in the Prokaryotic Expression System and the Study of the Aggregate of Its Biochemical Properties Relevant for the Cheese-Making Industry

AU - Belenkaya, S. V.

AU - Shcherbakov, D. N.

AU - Balabova, D. V.

AU - Belov, A. N.

AU - Koval, A. D.

AU - Elchaninov, V. V.

N1 - Publisher Copyright: © 2020, Pleiades Publishing, Inc. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2020/11

Y1 - 2020/11

N2 - The paper reports on the production of the maral recombinant chymosin in the Escherichia coli expression system (SHaffle express strain) and the study of its biochemical properties relevant for the cheese-making industry. The highest maral recombinant prochymosin content in inclusion bodies was observed when producer cells were cultured at 25°C for 6 h after the introduction of an inducer, 10 mM isopropyl-β-D-1-thiogalactopyranoside. The biochemical properties of the obtained enzyme were compared with those of bovine and dromedarian recombinant chymosins. It is shown that total proteolytic activity of the maral recombinant chymosin was comparable with that of the bovine enzyme and that it exceeded the activity of the dromedarian enzyme by about 3.8 times. The thermal stability of the recombinant chymosin from maral was found to be 5–10°C higher than that of the chymosins from the cow and dromedary. The patterns of dependence of milk-clotting activity on the pH level and calcium chloride concentration in cow’s milk on the enzyme from maral met the requirements set by the cheese industry. Its high proteolytic activity and thermal stability limits the scope of application of maral recombinant chymosin to the production of cheeses with short ripening and short storage times.

AB - The paper reports on the production of the maral recombinant chymosin in the Escherichia coli expression system (SHaffle express strain) and the study of its biochemical properties relevant for the cheese-making industry. The highest maral recombinant prochymosin content in inclusion bodies was observed when producer cells were cultured at 25°C for 6 h after the introduction of an inducer, 10 mM isopropyl-β-D-1-thiogalactopyranoside. The biochemical properties of the obtained enzyme were compared with those of bovine and dromedarian recombinant chymosins. It is shown that total proteolytic activity of the maral recombinant chymosin was comparable with that of the bovine enzyme and that it exceeded the activity of the dromedarian enzyme by about 3.8 times. The thermal stability of the recombinant chymosin from maral was found to be 5–10°C higher than that of the chymosins from the cow and dromedary. The patterns of dependence of milk-clotting activity on the pH level and calcium chloride concentration in cow’s milk on the enzyme from maral met the requirements set by the cheese industry. Its high proteolytic activity and thermal stability limits the scope of application of maral recombinant chymosin to the production of cheeses with short ripening and short storage times.

KW - calcium chloride concentration

KW - cheese-making

KW - milk-clotting activity

KW - pH

KW - proteolytic activity

KW - recombinant chymosin from maral

KW - thermal stability

KW - CAMEL CHYMOSIN

KW - CLONING

KW - CURD

KW - PROTEINS

UR - http://www.scopus.com/inward/record.url?scp=85096971485&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/22e15c1e-b6ef-3064-b28e-971d900b9ec1/

U2 - 10.1134/S0003683820060034

DO - 10.1134/S0003683820060034

M3 - Article

AN - SCOPUS:85096971485

VL - 56

SP - 647

EP - 656

JO - Applied Biochemistry and Microbiology

JF - Applied Biochemistry and Microbiology

SN - 0003-6838

IS - 6

ER -