Barnaul, Russian Federation
Barnaul, Russian Federation
Barnaul, Russian Federation
Barnaul, Russian Federation
Barnaul, Russian Federation
Barnaul, Russian Federation
employee
Barnaul, Barkaul, Russian Federation
The thermal stability and parameters of the Michaelis-Menten kinetics of engineered variant of recombinant reindeer chymosin (Rangifer tarandus) with a point amino acid substitution Lys53→Glu were studied. The values of the threshold of thermal inactivation of the studied enzyme and commercial recombinant cow chymosin coincided and amounted to 50 °C. The engineered reindeer enzyme showed the greatest affinity to the used chromogenic substrate, its Michaelis constant (Km) was 1,5–4,7 times lower than the same indicator of commercial recombinant chymosins. The catalytic rate constants (kcat) of the engineered variant of deer chymosin and recombinant camel chymosin were comparable. In terms of specificity (kcat/Km), the chymosins of reindeer and cow surpassed the genetically engineered analogue of the chymosin of the single-humped camel by 1,7 times.
chymosin, reindeer, engineered variant of recombinant chymosin, thermal stability, parameters of Michaelis-Menten kinetics
1. Bansal, N. Factors affecting the retention of rennet in cheese curd / N. Bansal, P. F. Fox, P. L. McSweeney // J. Agric. Food. Chem. 2007. V. 55 (22). P. 9219-9225.
2. Bansal, N. Comparison of the level of residual coagulant activity in different cheese varieties / N. Bansal, P. F. Fox, P. L. McSweeney // J. Dairy Res. 2009. V. 76. P. 290-293.
3. D`Incecco, P. Impact of Extending Hard-Cheese Ripening: A Multiparameter Characterization of Parmigiano Reggiano Cheese Ripened up to 50 Months / P. D»Incecco [et al.] // Foods. 2020. V. 9. P. 268.
4. Masotti, F. Proteolysis indices related to cheese ripening and typicalness in PDO Grana Padano cheese / F. Masotti [et al.] // Int. Dairy J. 2010. V. 20. № 5. P. 352-359.
5. Mane, A. Proteolysis in Irish farmhouse Camembert cheese during ripening / A. Mane, P. L. H. McSweeney // J. Food Biochem. 2020. V. 44. № 1.
6. Gumus, P. Effects of blends of camel and calf chymosin on proteolysis, residual coagulant activity, microstructure, and sensory characteristics of Beyaz peynir / P. Gumus, A. A. Hayaloglu // J. Dairy Sci. 2019. V. 102. N. 7. P. 5945-5956.
7. Lamichhane, P. Microstructure and fracture properties of semi-hard cheese: Differentiating the effects of primary proteolysis and calcium solubilization / P. Lamichhane [et al.] // Food Res Int. 2019. V. 125. P. 108-525.
8. Kappeler, S. R. Characterization of recombinant camel chymosin reveals superior properties for the coagulation of bovine and camel milk / S. R. Kappeler [et al.] // Biochem. Biophys. Res. Comm. 2006. V. 2. N. 342. P. 647-654.
9. Vallejo, J. A. Short communication: A comparative analysis of recombinant chymosins / J. A. Vallejo [et al.] // J. Dairy Sci. 2012. V. 95 (2). P. 609-613.
10. Jensen, J. L. The function of the milk-clotting enzymes bovine and camel chymosin studied by a fluorescence resonance energy transfer assay / J. L. Jensen [et al.] // J. Dairy Sci. 2015. V. 98. P. 2853-2860.
11. Pushkarev, V. A. Molokosvertyvayuschaya i obschaya proteoliticheskaya aktivnost' inzhenernogo varianta rekombinantnogo himozina severnogo olenya (Rangifer tarandus) / V. A. Pushkarev [i dr.] // Syrodelie i maslodelie. 2023. № 1. S. 22-25.
12. Pushkarev, V. A. Vliyanie koncentracii ionov kal'ciya i rN na molokosvertyvayuschuyu aktivnost' inzhenernogo varianta rekombinantnogo himozina severnogo olenya (Rangifer tarandus) / V. A. Pushkarev [i dr.] // Syrodelie i maslodelie. 2023. № 2. S. 31.
13. El'chaninov, V. V. Issledovanie molokosvertyvayuschego fermenta iz sychugov se¬vernyh oleney: dis. … kand. tehn. nauk: 05.18.04: zaschischena 23.01.06: utv. 07.04.06 / El'chaninov Vadim Valentinovich. - Kemerovo, 2006. - 172 s. - Bibliogr.: S. 121-147.
14. Belenkaya, S. V. Characterization of the Altai Maral Chymosin Gene, Production of a Chymosin Recombinant Analog in the Prokaryotic Expression System, and Analysis of Its Several Biochemical Properties / S. V. Belenkaya [et al.] // Biochemistry (Moscow). 2020. V. 85. № 7. P. 781-791.
15. Johnson, K. A. The original Michaelis constant: translation of the 1913 Michaelis - Menten paper / K. A. Johnson, R. S. Goody // Biochemistry. 2011. V. 50 (39). P. 8264-8269.
16. Belenkaya, S. V. Basic Biochemical Properties of Recombinant Chymosins (Review) / S. V. Belenkaya [et al.] // Appl. Biochem. Microbiol. 2020. V. 56. № 4. P. 363-372.
