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Statistical Optimization of Milk Clotting Enzyme Biosynthesis by Mucor mucedo Kp736529 and its Further Application in Cheese Production
Open accessAbstract
Microbial sources of bioagents are coinciding with the attempts to find cheap alternative substrates for high production.Rennet is the golden element in the dairy industry, particularly cheese making.Commonly, the use of statistical models to optimize culture medium components and conditions has increased in present-day biotechnology, due to its ready applicability and aptness.The present study adopted statistical approach for Milk Clotting Enzyme (MCE) production by Mucor mucedo KP736529 which was isolated, selected and molecularly identified, as the most potent active strain in MCE biosynthesis.Screening of various medium constituents using the statistical Plackett-Burman design showed glucose and KH PO as the significant factors affecting 2 4 MCE biosynthesis.Optimizing MCE production by the central composite design led to 6.12 fold increase in MCE production, compared with initial activity.Domiati cheese was manufactured using the M. mucedo KP736529 enzyme (E-cheese) or using commercial calf rennet (C-cheese) as control.During two months under pickling conditions, the yield and chemical properties of E-cheese were better than those of C-cheese.The fungal enzyme (MCE) showed proteolytic activity higher than calf rennet with the absence of taste defects such as bitter taste.Organoleptically, E-cheese scored the highest points.
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The availability of rennets is gradually decreasing, primarily because of rising demand for cheese production and increasing consumption.
“The availability of rennets is gradually decreasing, primarily because of the rising demand for cheese production and increasing consumption (Ayana et al., 2015;Ahmed et al., 2016).”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Ayana et al., 2015
The Introduction (page not provided) says animal-derived coagulants cannot meet cheese-production demand and describes mammalian rennet as having limited availability. It does not say rennet availability is gradually decreasing or link such a decline to increasing consumption. (LLM, conf=0.84)
Experimental — accuracy not yet measuredBitterness in cheese primarily results from the buildup of small hydrophobic peptides formed through the action of coagulants, especially rennet alternatives or microbial enzymes.
“The high MCA/PA ratio results in the formation of short peptides, which impart sour and bitter flavors, with bitterness in cheese primarily resulting from the buildup of small hydrophobic peptides formed through the action of coagulants, especially rennet alternatives or microbial enzymes (Ayana et al., 2015;D'Incecco et al., 2022;Di Rosa et al., 2024).”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Ayana et al., 2015
The Organoleptic evaluation section (page unavailable) says microbial rennet may cause bitterness during storage, while the Additional information section (page unavailable) describes the fungal enzyme’s greater proteolytic activity. Neither section identifies a buildup of small hydrophobic peptides as the primary cause of bitterness. The study also reports no bitterness in the cheese made with its fungal enzyme. (LLM, conf=0.86)
Experimental — accuracy not yet measuredA high MCA/PA ratio results in the formation of short peptides, which impart sour and bitter flavors.
“The high MCA/PA ratio results in the formation of short peptides, which impart sour and bitter flavors, with bitterness in cheese primarily resulting from the buildup of small hydrophobic peptides formed through the action of coagulants, especially rennet alternatives or microbial enzymes (Ayana et al., 2015;D'Incecco et al., 2022;Di Rosa et al., 2024).”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Ayana et al., 2015
The Organoleptic evaluation section says microbial rennet might cause bitterness during storage, but it does not link bitterness or sourness to short peptides or a high MCA/PA ratio. It also reports no bitterness or flavor defects in the cheese made with the studied fungal enzyme. The Additional information section discusses protein degradation into peptides, without establishing the claimed ratio-to-flavor relationship; page numbers are unavailable in our copy. (LLM, conf=0.84)
Experimental — accuracy not yet measuredThere is a growing number of scientific reports on the production and extraction of new alternatives for milk-clotting enzymes, driven by growing demand for cheese and limited availability of rennet.
“There is a growing number of scientific reports on the production and extraction of new alternatives for milk-clotting enzymes (MCE), as a result of the growing demand for cheese and the limited availability of rennet.”
From Invited review: Milk-clotting enzymes of microbial origin and their role in cheesemaking-A review. (2025) · cites this paper as Ayana et al., 2015
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References 10 · Cited by 18
Metadata
- Authors
- Ayana, I. A. A. A., Ibrahim, A. E., & Saber, W. I. A.
- Affiliations
- Agricultural Research Center (EG)
- Venue
- International Journal of Dairy Science
- Publisher
- Science Alert
- Published
- 2015-02-15
- Volume
- 10
- Issue
- 2
- Pages
- 61-76
- ISSN
- 1811-9743 (ISSN-L)
- OpenAlex
- W2546458532
- Cited by
- 18
- References
- 10
- Citations by year
- 2025: 1 · 2023: 3 · 2022: 3 · 2021: 2 · 2020: 4 · 2019: 1 · 2018: 1 · 2017: 1 · 2016: 2
- FWCI
- 1.11
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- International Journal of Dairy Science (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Agricultural and Biological Sciences › Food Science › Advanced Scientific Research Methods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
- Keywords
- milk-clotting enzyme, Mucor mucedo, cheese production, fungal proteases, Plackett-Burman design, central composite design, culture medium optimization, Domiati cheese, proteolytic activity, cheese sensory evaluation
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