Mujica MÁ, Matute-Almeida L, Rodas-Espinoza S, Román-Rojas H, Vargas-Guzmán D

Language: Spanish
References: 26
Page: 3201-3207
PDF size: 325.18 Kb.

ABSTRACT

Chymosin is a protease (EC 3.4.23.4) that is required in the manufacture of cheese. Due to the high production costs of this enzyme, its obtention has been investigated in microbial fermentation processes, such as those generated by Rhizomucor miehei. Researchers have pointed out that this enzyme has a high milk coagulation activity and its production is affected by some operational parameters. Therefore, in this work, the effect of the agitation speed, the initial pH of the culture medium and the fermentation time in obtaining the microbial rennet produced by Rhizomucor miehei was studied. For this, a factorial 22 design completely random was selected, the initial pH (4.0 and 6.0), the agitation speed (200 and 300 rpm), the variation of pH, the glucose consumption and the enzymatic activity at 0, 24, 48 and 72 hours of fermentation were determined. The results showed that the highest enzymatic activity was obtained after 72 hours of fermentation with an initial pH of 4.0 and agitation of 300 rpm (p < 0.05). Likewise, it was determined that the interaction of the agitation speed, initial pH and fermentation time had a highly significant influence on the enzymatic activity (p < 0.01) and that there is no linear correlation between the rate of glucose consumption in the medium and the enzymatic activity obtained. Finally, it was observed that supplementing the medium with lactose decreased the coagulant activity of the milk’s protease enzyme.

REFERENCES

1. García Garibay M, Quintero Ramírez R, Lóez-Mungía Canales A. Biotecnología Alimentaria. México: Editorial Limusa, S.A.; 1999.

2. Morillo Piña OT, García Lugo PJ, Ruiz B, Guerrero Cardena G, Torres Vielma Y, Castañeda Ruiz RF. Evaluación de la producción experimental de enzimas coagulantes de leche utilizando cepas de Rhizomucor spp. Rev Colombiana Biotecnol. 2015;17(1):54-60.

3. Merheb-Dini C, Gomes E, Boscolo M, Da Silva R. Production and characterization of a milk-clotting protease in the crude enzymatic extract from the newly isolated Thermomucorindicae-seudaticae N31 (Milk-clotting protease from the newly isolated Thermomucorindicae-seudaticaeN31). Food Chem. 2010;120:87-93.

4. Cheng Q, Bai X, Wang Y. Isolation and fermentation condition of milkclotting enzyme producing strain from glutinous rice wine. Chinese J Biotechnol. 2008; 24(6):999-1003.

5. Khademi, F, Abachi S, Mortazavi A, Ehsani MA, Tabatabaei MR, Malekzadeh FA. Optimization of fungal rennet production by local isolate of Rhizomucor nainitalensis. Under Solid Substrate Fermentation system. IOSR J Pharm Biol Sci. 2013;5(2):115-21.

6. Silveira G, Moneteiro de Oliveira G, Ribeiro EJ, Monti R, Contiero J. Microbial rennet produced by Mucor miehei in solidstate and submerged fermentation. Braz Arch Biol Technol. 2005;48(6):931-7.

7. Nouani A, Belhamiche N, Slamani R, Belbraquet S, Fazouane F, Bellal MM. Extracellular protease from Mucor pusillus: purification and characterization. Int J Dairy Technol. 2009;62(1):112-7.

8. AmerAbeer EA, Hashem MI, Amer ME, Gomaa AM. Using sweet whey for production of milk clotting enzyme by Mucor miehei NRRL 3420 in production of white soft cheese. Middle East J Appl Sci. 2015;5(4):1068-81.

9. Kazemi-Vaysari A, Kheirolomoom A, Arjmand M, Habibollahi M. Optimization of Mucor-miehei rennin production and recovery. Scientia Iranica. 2002;9(1):99-104.

10. Matute L, Bertsch A, Díaz I. Evaluación de la actividad amilolítica de Aspergillus niger ANM-1 en fermentaciones en estado sólido y sumergido para la obtención y caracterización de aditivos enzimáticos. Rev Fac Agron Universidad Central de Venezuela. 2012;38(1):9-17.

11. Sánchez-Henao CP, Escobar Kousen JM, Rodríguez de Stouvenel A. Características de la producción de la renina microbiana de Mucormiehei en un proceso de alimentación por lote. Rev Colombiana Biotecnol. 1999;2(1):28-34.

12. Miller GL. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem. 1959;31(3):426-8.

13. Otani H, Mitsuhiro I, Akiyoshi H. The screening of trees having milk activity. Animal Sci Technol. 1991;62:417-23.

14. Kawai M, Mukai N. Studies on milkclotting enzyme produced by Basidomycetes. 1. Screening tests of Basidiomycetes for the production of milk clotting enzymes. Agric Biol Chem. 1970;34:159-63.

15. Yasni S, Maulidya A. Development of Corn Milk Yoghurt Using Mixed Culture of Lactobacillus delbruekii, Streptococcus salivarus, and Lactobacillus casei. J Biosci. 2014;21(1):1-7.

16. Association of Analytical Communities. AOAC 931.12. Refractometría. In: 16. Association of Analytical Communities International. Official methods of analysis. (18.a ed.). 2005.

17. Camacho-Ruiz L, Pérez-Guerra N, Pérez Roses R. Factors affecting the growth of Saccharomyces cerevisiae in batch culture and in solid state fermentation. Electron J Environ Agric Food Chem. 2003;2:531-42.

18. Gutiérrez H, de la Vara R. Análisis y Diseño de Experimentos. Segunda edición. México: McGraw-Hill Interamericana; 2008.

19. IBM Corp. Released. IBM SPSS Statistics for Windows, Version 20.0. Armonk, New York: IBM Corporation; 2011.

20. Yuqiu L, Hua T, Da L, Zhoulin L, Yanping C, Yuanyuan J, et al. Screening and characterization of a mutant fungal aspartic proteinase from Mucor pusillus. Open Biotechnol J. 2015;9:119-26.

21. Areces L, Biscoglio de Jiménez M, Parry M, Fraile E, Fernández H, Cascone O. Purification and characterization of a milk clotting protease from Mucor bacilliformis. Appl Biochem Bio-technol. 1992;37:283-94.

22. Thakur MS, Karanth NG, Nand K. Production of fungal rennet by Mucor miehei using solid state fermentation. Appl Microbiol Biotechnol. 1990;32:409-13.

23. Escobar J, Barnnet S. Effect of agitation speed on the synthesis of Mucor miehei acid protease. Enzyme Microbiol Technol. 1993;15:1009-13.

24. Alves MH, de Campos-Takaki G, Okada K, Ferreira Pessoa I, Milanez AI. Detection of extracelular protease in Mucor species. Rev Iberoam Micol. 2005;22:114-7.

25. De Lima CJB, Cortez M, Lovaglio RB, Ribeiro EJ, Contiero J, De Araújo EH. Production of rennet in submerged fermentation with the filamentous fungus Mucor miehei NRRL 3420. World Appl Sci J. 2008;4(4):578-85.

26. Rocha da Silva C, BoechatDelatorre A, Leal Martins ML. Effect of the culture conditions on the production of an extracellular protease by thermophilic Bacillus sp and some properties of the enzymatic actibity. Braz J Microbiol. 2007;38:253-8.