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Can Recombinant Tree Shrew (Tupaia belangeri chinensis) Chymosin Coagulate Cow (Bos taurus) Milk? / Balabova, D. V.; Belenkaya, S. V.; Volosnikova, E. A. et al.

In: Applied Biochemistry and Microbiology, Vol. 58, No. 6, 12.2022, p. 761-770.

Research output: Contribution to journal › Article › peer-review

Harvard

Balabova, DV, Belenkaya, SV, Volosnikova, EA, Hermes, T, Chirkova, VY, Sharlaeva, EA, Shcherbakov, DN, Belov, AN, Koval, AD & Elchaninov, VV 2022, 'Can Recombinant Tree Shrew (Tupaia belangeri chinensis) Chymosin Coagulate Cow (Bos taurus) Milk?', Applied Biochemistry and Microbiology, vol. 58, no. 6, pp. 761-770. https://doi.org/10.1134/S0003683822060023

APA

Balabova, D. V., Belenkaya, S. V., Volosnikova, E. A., Hermes, T., Chirkova, V. Y., Sharlaeva, E. A., Shcherbakov, D. N., Belov, A. N., Koval, A. D., & Elchaninov, V. V. (2022). Can Recombinant Tree Shrew (Tupaia belangeri chinensis) Chymosin Coagulate Cow (Bos taurus) Milk? Applied Biochemistry and Microbiology, 58(6), 761-770. https://doi.org/10.1134/S0003683822060023

Vancouver

Balabova DV, Belenkaya SV, Volosnikova EA, Hermes T, Chirkova VY, Sharlaeva EA et al. Can Recombinant Tree Shrew (Tupaia belangeri chinensis) Chymosin Coagulate Cow (Bos taurus) Milk? Applied Biochemistry and Microbiology. 2022 Dec;58(6):761-770. doi: 10.1134/S0003683822060023

Author

Balabova, D. V. ; Belenkaya, S. V. ; Volosnikova, E. A. et al. / Can Recombinant Tree Shrew (Tupaia belangeri chinensis) Chymosin Coagulate Cow (Bos taurus) Milk?. In: Applied Biochemistry and Microbiology. 2022 ; Vol. 58, No. 6. pp. 761-770.

BibTeX

@article{81277aec31f34aeab88e3a7f045e195c,
title = "Can Recombinant Tree Shrew (Tupaia belangeri chinensis) Chymosin Coagulate Cow (Bos taurus) Milk?",
abstract = "Genetically engineered chymosin from the tree shrew (Tupaia belangeri chinensis) has been obtained and partially characterized for the first time. The target enzyme was produced in Escherichia coli, strain BL21(DE3). It was shown that tree shrew recombinant chymosin coagulates cow milk (Bos taurus). The total and specific milk-clotting activity of the obtained enzyme was 0.7–5.3 IMCU/mL and 8.8–16.6 IMCU/mg. The nonspecific proteolytic activity of tree shrew recombinant chymosin in relation to total bovine casein was 30 and 117% higher than that of recombinant chymosin of cow and of single-humped camel respectively. It was found that in comparison with most of the known genetically engineered chymosins, the tree shrew enzyme showed exceptionally low thermal stability. After heating at 45°C, the coagulation ability of tree shrew recombinant chymosin decreased by more than 40%, and at 50°C the enzyme lost more than 90% of the initial milk-clotting activity. The Michaelis constant (Km), enzyme turnover number (kcat), and catalytic efficiency (kcat/Km) for genetically engineered chymosin from the tree shrew were 6.3 ± 0.1 µM, 11 927 ± 3169 s–1 and 1968 ± 620 µM–1 s–1, respectively. Comparative analysis showed that the primary structure of the chymosin-sensitive site of cow kappa-casein and the supposed similar sequence of tree shrew kappa-casein differed by 75%. The ability of tree shrew recombinant chymosin to coagulate cow{\textquoteright}s milk, along with a low thermal stability and high catalytic efficiency with respect to the substrate, imitating the chymosin-sensitive site of cow kappa-casein, suggests that this enzyme is of potential interest for cheese making.",
keywords = "bovine milk, cheese making, milk-clotting activity, proteolytic activity, recombinant chymosin, thermal stability, tree shrew",
author = "Balabova, {D. V.} and Belenkaya, {S. V.} and Volosnikova, {E. A.} and T. Hermes and Chirkova, {V. Y.} and Sharlaeva, {E. A.} and Shcherbakov, {D. N.} and Belov, {A. N.} and Koval, {A. D.} and Elchaninov, {V. V.}",
note = "Funding Information: The work was carried out within the framework of the state task of the Ministry of Science and Higher Education of the Russian Federation (FZMW-2020-0002, Development of producers of recombinant enzymes for cheese making). Publisher Copyright: {\textcopyright} 2022, The Author(s).",
year = "2022",
month = dec,
doi = "10.1134/S0003683822060023",
language = "English",
volume = "58",
pages = "761--770",
journal = "Applied Biochemistry and Microbiology",
issn = "0003-6838",
publisher = "Maik Nauka-Interperiodica Publishing",
number = "6",

}

RIS

TY - JOUR

T1 - Can Recombinant Tree Shrew (Tupaia belangeri chinensis) Chymosin Coagulate Cow (Bos taurus) Milk?

AU - Balabova, D. V.

AU - Belenkaya, S. V.

AU - Volosnikova, E. A.

AU - Hermes, T.

AU - Chirkova, V. Y.

AU - Sharlaeva, E. A.

AU - Shcherbakov, D. N.

AU - Belov, A. N.

AU - Koval, A. D.

AU - Elchaninov, V. V.

N1 - Funding Information: The work was carried out within the framework of the state task of the Ministry of Science and Higher Education of the Russian Federation (FZMW-2020-0002, Development of producers of recombinant enzymes for cheese making). Publisher Copyright: © 2022, The Author(s).

PY - 2022/12

Y1 - 2022/12

N2 - Genetically engineered chymosin from the tree shrew (Tupaia belangeri chinensis) has been obtained and partially characterized for the first time. The target enzyme was produced in Escherichia coli, strain BL21(DE3). It was shown that tree shrew recombinant chymosin coagulates cow milk (Bos taurus). The total and specific milk-clotting activity of the obtained enzyme was 0.7–5.3 IMCU/mL and 8.8–16.6 IMCU/mg. The nonspecific proteolytic activity of tree shrew recombinant chymosin in relation to total bovine casein was 30 and 117% higher than that of recombinant chymosin of cow and of single-humped camel respectively. It was found that in comparison with most of the known genetically engineered chymosins, the tree shrew enzyme showed exceptionally low thermal stability. After heating at 45°C, the coagulation ability of tree shrew recombinant chymosin decreased by more than 40%, and at 50°C the enzyme lost more than 90% of the initial milk-clotting activity. The Michaelis constant (Km), enzyme turnover number (kcat), and catalytic efficiency (kcat/Km) for genetically engineered chymosin from the tree shrew were 6.3 ± 0.1 µM, 11 927 ± 3169 s–1 and 1968 ± 620 µM–1 s–1, respectively. Comparative analysis showed that the primary structure of the chymosin-sensitive site of cow kappa-casein and the supposed similar sequence of tree shrew kappa-casein differed by 75%. The ability of tree shrew recombinant chymosin to coagulate cow’s milk, along with a low thermal stability and high catalytic efficiency with respect to the substrate, imitating the chymosin-sensitive site of cow kappa-casein, suggests that this enzyme is of potential interest for cheese making.

AB - Genetically engineered chymosin from the tree shrew (Tupaia belangeri chinensis) has been obtained and partially characterized for the first time. The target enzyme was produced in Escherichia coli, strain BL21(DE3). It was shown that tree shrew recombinant chymosin coagulates cow milk (Bos taurus). The total and specific milk-clotting activity of the obtained enzyme was 0.7–5.3 IMCU/mL and 8.8–16.6 IMCU/mg. The nonspecific proteolytic activity of tree shrew recombinant chymosin in relation to total bovine casein was 30 and 117% higher than that of recombinant chymosin of cow and of single-humped camel respectively. It was found that in comparison with most of the known genetically engineered chymosins, the tree shrew enzyme showed exceptionally low thermal stability. After heating at 45°C, the coagulation ability of tree shrew recombinant chymosin decreased by more than 40%, and at 50°C the enzyme lost more than 90% of the initial milk-clotting activity. The Michaelis constant (Km), enzyme turnover number (kcat), and catalytic efficiency (kcat/Km) for genetically engineered chymosin from the tree shrew were 6.3 ± 0.1 µM, 11 927 ± 3169 s–1 and 1968 ± 620 µM–1 s–1, respectively. Comparative analysis showed that the primary structure of the chymosin-sensitive site of cow kappa-casein and the supposed similar sequence of tree shrew kappa-casein differed by 75%. The ability of tree shrew recombinant chymosin to coagulate cow’s milk, along with a low thermal stability and high catalytic efficiency with respect to the substrate, imitating the chymosin-sensitive site of cow kappa-casein, suggests that this enzyme is of potential interest for cheese making.

KW - bovine milk

KW - cheese making

KW - milk-clotting activity

KW - proteolytic activity

KW - recombinant chymosin

KW - thermal stability

KW - tree shrew

UR - http://www.scopus.com/inward/record.url?scp=85143292678&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/7b6642ad-46c2-322b-885e-9a205b654e88/

U2 - 10.1134/S0003683822060023

DO - 10.1134/S0003683822060023

M3 - Article

AN - SCOPUS:85143292678

VL - 58

SP - 761

EP - 770

JO - Applied Biochemistry and Microbiology

JF - Applied Biochemistry and Microbiology

SN - 0003-6838

IS - 6

ER -

ID: 40356292