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Production and Characterization of a Milk-clotting Protease Produced in Submerged Fermentation by the Thermophilic Fungus Thermomucor indicae-seudaticae N31

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Abstract

Proteases are some of the most important industrial enzymes, and one of their main applications is for the production of cheese in the dairy industry. Due to a shortage of animal rennet, microbial coagulant proteases are being sought. In this work, the production of microbial rennet from Thermomucor indicae-seudaticae N31 was studied in submerged fermentation. The best enzyme production was obtained in a fermentation medium containing 4 % wheat bran as the substrate in 0.3 % saline solution, incubated for 72 h at 45 °C and 150 rpm. The value of the milk clotting activity (MCA) was 60.5 U/mL, and the ratio to proteolytic activity (MCA/PA) was 510. The crude enzyme showed optimum pH at 5.5 and two peaks of optimum temperature (MCA at 65 °C and PA at 60 °C). The MCA was stable in the pH range 4.0–4.5 for 24 h and up to 55 °C for 1 h. It was stable during storage at different temperatures (−20 to 25 °C) for 10 weeks. Based on these results, we conclude that microbial rennet from T. indicae-seudaticae N31 produced by submerged fermentation showed good prospects of replacing traditional rennet.

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Acknowledgments

We acknowledge the financial support for carrying out this study which was provided by CAPES and FAPESP (Brazil).

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Authors and Affiliations

  1. Laboratory of Biochemistry and Applied Microbiology, São Paulo State University–UNESP, Rua Cristóvão Colombo, 2265, São Jose do Rio Preto, São Paulo, 15054-000, Brazil

    B. L. Silva, F. M. Geraldes, C. S. Murari, E. Gomes & R. Da-Silva

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  1. B. L. Silva
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  2. F. M. Geraldes
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  3. C. S. Murari
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  4. E. Gomes
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  5. R. Da-Silva
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Correspondence to R. Da-Silva.

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Silva, B.L., Geraldes, F.M., Murari, C.S. et al. Production and Characterization of a Milk-clotting Protease Produced in Submerged Fermentation by the Thermophilic Fungus Thermomucor indicae-seudaticae N31. Appl Biochem Biotechnol 172, 1999–2011 (2014). https://doi.org/10.1007/s12010-013-0655-7

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  • DOI: https://doi.org/10.1007/s12010-013-0655-7

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