TY - JOUR

T1 - Biochemical and technological properties of moose (Alces alces) recombinant chymosin

AU - Balabova, D. V.

AU - Rudometov, A. P.

AU - Belenkaya, S. V.

AU - Belov, A. N.

AU - Koval, A. D.

AU - Bondar, A. A.

AU - Bakulina, A. Yu

AU - Rukhlova, E. A.

AU - Elchaninov, V. V.

AU - Shcherbakov, D. N.

N1 - Sanger DNA sequencing was performed in Genomics Core Facility (ICBFM SB RAS, Novosibirsk). This study was funded by the State Task of the Ministry of Science and Education of the Russian Federation (topic No. FZMW-2020-0002, “Design of recombinant enzyme producers for the cheese-making industry”) and The Russian Foundation for Basic Research (grant number 19-44-220010). Publisher Copyright: © 2022 Institute of Cytology and Genetics of Siberian Branch of the Russian Academy of Sciences. All rights reserved.

PY - 2022/5

Y1 - 2022/5

N2 - Recombinant chymosins (rСhns) of the cow and the camel are currently considered as standard milk coagulants for cheese-making. The search for a new type of milk-clotting enzymes that may exist in nature and can surpass the existing “cheese-making” standards is an urgent biotechnological task. Within this study, we for the first time constructed an expression vector allowing production of a recombinant analog of moose chymosin in the expression system of Escherichia coli (strain SHuffle express). We built a model of the spatial structure of moose chymosin and compared the topography of positive and negative surface charges with the correspondent structures of cow and camel chymosins. We found that the distribution of charges on the surface of moose chymosin has common features with that of cow and camel chymosins. However, the moose enzyme carries a unique positively charged patch, which is likely to affect its interaction with the substrate. Biochemical and technological properties of the moose rChn were studied. Commercial rСhns of cow and camel were used as comparison enzymes. In some technological parameters, the moose rChn proved to be superior to the reference enzymes. Сompared with the cow and camel rСhns, the moose chymosin specific activity is less dependent on the changes in CaCl2 concentration in the range of 1-5 mM and pH in the range of 6-7, which is an attractive technological property. The total proteolytic activity of the moose rСhn occupies an intermediate position between the rСhns of cow and camel. The combination of biochemical and technological properties of the moose rСhn argues for further study of this enzyme.

AB - Recombinant chymosins (rСhns) of the cow and the camel are currently considered as standard milk coagulants for cheese-making. The search for a new type of milk-clotting enzymes that may exist in nature and can surpass the existing “cheese-making” standards is an urgent biotechnological task. Within this study, we for the first time constructed an expression vector allowing production of a recombinant analog of moose chymosin in the expression system of Escherichia coli (strain SHuffle express). We built a model of the spatial structure of moose chymosin and compared the topography of positive and negative surface charges with the correspondent structures of cow and camel chymosins. We found that the distribution of charges on the surface of moose chymosin has common features with that of cow and camel chymosins. However, the moose enzyme carries a unique positively charged patch, which is likely to affect its interaction with the substrate. Biochemical and technological properties of the moose rChn were studied. Commercial rСhns of cow and camel were used as comparison enzymes. In some technological parameters, the moose rChn proved to be superior to the reference enzymes. Сompared with the cow and camel rСhns, the moose chymosin specific activity is less dependent on the changes in CaCl2 concentration in the range of 1-5 mM and pH in the range of 6-7, which is an attractive technological property. The total proteolytic activity of the moose rСhn occupies an intermediate position between the rСhns of cow and camel. The combination of biochemical and technological properties of the moose rСhn argues for further study of this enzyme.

KW - Alces alces

KW - biochemical properties

KW - cheese-making

KW - milk-clotting activity

KW - moose

KW - recombinant chymosin

KW - Alces alces

KW - biochemical properties

KW - cheese-making

KW - milk-clotting activity

KW - moose

KW - recombinant chymosin

UR - http://www.scopus.com/inward/record.url?scp=85133094249&partnerID=8YFLogxK

UR - https://www.elibrary.ru/item.asp?id=48556101

U2 - 10.18699/VJGB-22-31

DO - 10.18699/VJGB-22-31

M3 - Article

C2 - 35774365

AN - SCOPUS:85133094249

VL - 26

SP - 240

EP - 249

JO - Вавиловский журнал генетики и селекции

JF - Вавиловский журнал генетики и селекции

SN - 2500-0462

IS - 3

M1 - 3

ER -