Evaluation of the possibility of using whey to produce proteases of thermophilic bacteria of Bacillus genus
- Authors: Mirzalieva N.A.1, Beloded A.V.1, Romanova M.V.1
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Affiliations:
- D. Mendeleev University of Chemical Technology
- Issue: Vol 28, No 9 (2025)
- Pages: 39-44
- Section: Biological chemistry
- URL: https://journals.eco-vector.com/1560-9596/article/view/690136
- DOI: https://doi.org/10.29296/25877313-2025-09-05
- ID: 690136
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Abstract
Introduction. Proteolytic enzymes have great potential for introduction into modern industrial technologies, medical and scientific practice, agriculture and other spheres of human activity. Thermophilic bacteria are promising producers of proteases, since they are characterized by a wide variety, easy cultivation and the ability to use inexpensive substrates compared to sources of plant and animal origin, and the enzymes produced by thermophiles have high stability at elevated temperatures and extreme pH values. Milk whey is characterized by the presence of proteins, carbohydrates, and minerals, which makes it a valuable substrate for the cultivation of microorganisms.
The purpose of research was to assess the ability of the use of low-fat whey to obtain proteolytic enzymes of thermophilic strain Bacillus velezensis Kb.1.Gl.8.
Material and methods. Thermophilic strain Bacillus velezensis Kb.1.Gl.8 was used as a protease producer. The composition of the whey (i.e. proteins and carbohydrates contents) was determined by Lowry and Kjeldahl methods and HPLC. An analysis of the proteolytic activity of the culture supernatant was performed using casein as a substrate. To determine molecular weights of the proteolytic enzymes we used a zymographic analysis.
Results. The concentration of proteins in the whey was 7.11 g/L, crude protein – 10.63 g/L, lactose – 47.01 g/L. When culturing the thermophilic strain Bacillus velezensis Kb.1.Gl.8. on the low-fat whey, proteolytic activity of the enzymes in supernatant was 778.6 U/mL after 24 h and 212.2 U/mL after 48 h. The proteolytic activity of thermophilic bacteria grown on whey was higher than proteolytic activity on LB media. According to zymography, enzymes have molecular weights of 17, 32, 35, 55, 75 kDa.
Conclusions. Whey is a promising raw material for the cultivation of bacteria to produce enzymes with high yield. Further research may be focused on optimizing the nutrient media composition and growth conditions to increase the yield of proteases.
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About the authors
N. A. Mirzalieva
D. Mendeleev University of Chemical Technology
Author for correspondence.
Email: mirzalievanargiz02@gmail.com
ORCID iD: 0009-0000-7076-1794
Master`s Degree student of the Department of Biotechnology
Russian Federation, Geroyev Panfilovtsev Str., 20, Moscow, 125480A. V. Beloded
D. Mendeleev University of Chemical Technology
Email: beloded.a.v@muctr.ru
ORCID iD: 0000-0002-4425-8068
SPIN-code: 2852-6740
Ph.D. (Biol.), Associate Professor of the Department of Biotechnology
Russian Federation, Geroyev Panfilovtsev Str., 20, Moscow, 125480M. V. Romanova
D. Mendeleev University of Chemical Technology
Email: romanova.m.v@muctr.ru
ORCID iD: 0000-0003-3109-8445
SPIN-code: 6777-7438
Post-graduate Student, Assistant of the Department of Biotechnology
Russian Federation, Geroyev Panfilovtsev Str., 20, Moscow, 125480References
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