Production of highly active fungal milk-clotting enzyme by solid-state fermentation
Abstract
Cheese production is projected to reach 20 million metric tons by 2020, of which 33% is being produced using calf rennet (EC 3.4.23.4). There is shortage of calf rennet, and use of plant and microbial rennets, hydrolyze milk proteins non-specifically resulting in low curd yields. This study reports fungal enzymes obtained from cost effective medium, with minimal down streaming, whose activity is comparable with calf and Mucor rennet. Of the fifteen fungi that were screened, Mucor thermohyalospora (MTCC 1384) and Rhizopus azygosporus (MTCC 10195) exhibited the highest milk-clotting activity (MCA) of 18,383 ± 486 U/ml and 16,373 ± 558 U/ml, respectively. Optimization exhibited a 33% increase in enzyme production (30 g wheat bran containing 6% defatted soy meal at 30 °C, pH 7) for M. thermohyalospora. The enzyme was active from pH 5–10 and temperature 45–55 °C. Rhizopus azygosporus exhibited 31% increase in enzyme production (30 g wheat bran containing 4% defatted soy meal at 30 °C, pH 6) and the enzyme was active from pH 6–9 at 50 °C. Curd yields prepared from fungal enzyme extract decreased (5–9%), when compared with calf rennet and Mucor rennet. This study describes the potential of fungal enzymes, hitherto unreported, as a viable alternative to calf rennet
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Rhizopus azygosporus exhibits higher protease and lipase activities during milk clotting.
“Chinmayee et al. (2019) demonstrated that Rhizopus azygosporus exhibits higher protease and lipase activities during milk clotting.”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Chinmayee et al., 2019
Curd yield obtained using fungal R. azygosporus enzyme extract was 5% to 9% lower compared with that obtained with calf rennet and Mucor thermohyalospora.
“Chinmayee et al. (2019) showed that curd yield obtained using fungal R. azygosporus enzyme extract was 5% to 9% lower compared with that obtained with calf rennet and Mucor thermohyalospora.”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Chinmayee et al., 2019
Most microbiological MCE have high PA and low MCA/PA, which results in a lower cheese yield and bitter taste.
“Most microbiological MCE have high PA and low MCA/PA, which results in a lower cheese yield and bitter taste (Chinmayee et al., 2019;Liu et al., 2021).”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Chinmayee et al., 2019
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- Authors
- Chinmayee, C. V., Vidya, C., Rani, A., & Singh, S. A.
- Affiliations
- Central Food Technological Research Institute (IN)
- Department of Protein Chemistry and Technology, CSIR-Central Food Technological Research Institute, Mysuru, India;
- Corresponding author
- Singh, Sridevi Annapurna
- Venue
- Preparative Biochemistry & Biotechnology
- Publisher
- Informa UK Limited
- Published
- 2019-06-20
- Volume
- 49
- Issue
- 9
- Pages
- 858-867
- ISSN
- 1082-6068 (ISSN-L)
- 1532-2297
- OpenAlex
- W2953038309
- Cited by
- 12
- References
- 40
- Citations by year
- 2026: 1 · 2025: 3 · 2023: 3 · 2022: 1 · 2021: 4
- FWCI
- 0.69
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Preparative Biochemistry & Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Funders
- University Grants Commission
- Department of Science and Technology, Govt. of India
- 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
- milk-clotting enzyme, solid-state fermentation, cheese production, curd yield, wheat bran substrate, enzyme production optimization, enzyme activity
- MeSH terms
- Fermentation, Hydrogen-Ion Concentration, Industrial Microbiology, Mucor, Rhizopus, Substrate Specificity, Temperature, Aspartic Acid Endopeptidases
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