Production and characterization of a milk-clotting protease produced in submerged fermentation by the thermophilic fungus Thermomucor indicae-seudaticae N31
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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The enzyme produced by Thermomucor spp. exhibited high activity at pH 4.0 to 4.5 for 24 h.
“The enzyme produced by Thermomucor spp.exhibited high activity at pH 4.0 to 4.5 for 24 h (Silva et al., 2014),”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Silva et al., 2014
The highest MCE production was achieved in a medium with 4% wheat bran.
“Silva et al. (2014) used the following products as carbon sources: wheat bran (with or without added casein), corn residues, flour, soy milk, and others, with the highest MCE production achieved in a medium with 4% wheat bran (Silva et al., 2014).”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Silva et al., 2014
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- Authors
- Silva, B. L., Geraldes, F. M., Murari, C. S., Gomes, E., & Da-Silva, R.
- Affiliations
- Universidade Estadual Paulista (Unesp) (BR)
- Corresponding author
- Da-Silva, R.
- Venue
- Applied Biochemistry and Biotechnology
- Published
- 2013-12-08
- Volume
- 172
- Issue
- 4
- Pages
- 1999-2011
- ISSN
- 0273-2289 (ISSN-L)
- 1559-0291
- OpenAlex
- W1981419970
- Cited by
- 35
- References
- 37
- Citations by year
- 2026: 1 · 2025: 3 · 2024: 2 · 2023: 4 · 2022: 1 · 2021: 6 · 2020: 1 · 2019: 2 · 2018: 3 · 2017: 4 · 2016: 6 · 2015: 1 · 2014: 1
- FWCI
- 2.34
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Applied Biochemistry and Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Acervo Digital da Universidade Estadual Paulista (Universidade Estadual Paulista) (submittedVersion)
- UNESP Institutional Repository (São Paulo State University) (submittedVersion)
- Funders
- Fundação de Amparo à Pesquisa do Estado de São Paulo
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
- Life Sciences › Agricultural and Biological Sciences › Food Science › Advanced Scientific Research Methods
- Keywords
- microbial rennet, submerged fermentation, thermophilic fungi, wheat bran substrate, cheese coagulation, enzyme characterization, enzyme stability, proteolytic activity
- MeSH terms
- Dietary Fiber, Fermentation, Industrial Microbiology, Mucorales, Peptide Hydrolases, Chymosin
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