DETERMINATION OF THE ACTIVITY LEVEL OF THE GALACTOSIDASE ENZYME ISOLATED FROM CHICKPEAS

522 Volume 5, Issue 10: Special Issue (EJAR) ISSN: 2181-2020 MPHAPP THE 6TH INTERNATIONAL SCIENTIFIC AND PRACTICAL CONFERENCE “MODERN PHARMACEUTICS: ACTUAL PROBLEMS AND PROSPECTS” TASHKENT, OCTOBER 17, 2025 in-academy.uz DETERMINATION OF THE ACTIVITY LEVEL OF THE GALACTOSIDASE ENZYME ISOLATED FROM CHICKPEAS Po’latova Khurriyat Tashkent Pharmaceutical Institute Uzbekistan, Tashkent, oybek 45 e-mail: [email protected], tel: +998-99 697-41-32 https://doi.org/10.5281/zenodo.17343868 Relevance: According to the Decree of the President of the Republic of Uzbekistan No. PF4947 dated February 7, 2017 "On the Strategy of Actions for the Further Development of the Republic of Uzbekistan", general goals are set, such as developing agriculture, ensuring food security, and increasing export potential. Within the framework of this strategy, measures are being implemented to expand and support the cultivation of legumes, including peas. Galactosidases are hydrolytic enzymes capable of breaking down galactose residues, which are found in many microorganisms, plants and animal cells. Their biological and industrial importance is currently increasing. In biotechnology, enzymes are widely used in processes such as bioconversion, synthesis of prebiotic oligosaccharides, and development of pharmaceuticals. The galactosidase enzyme is gaining great relevance in modern biotechnology, pharmaceuticals and the food industry. It plays an important role not only in eliminating lactose intolerance, but also in the production of bioactive substances with high added value. Therefore, obtaining this enzyme in a highly active form based on genetic modification and putting it into practice is one of the urgent tasks. Galactosidase, in particular β - galactosidase, is an important glycosidase that catalyzes the hydrolysis of β - galactosidase bonds in various carbohydrates, including oligosaccharides and polysaccharides. It plays a crucial role in lactose metabolism, facilitating its breakdown into glucose and galactose, thereby improving digestion, and is especially abundant in dairy products. Propose of study: It consists of determining the activity level of the galactosidase enzyme isolated from the chickpea plant. Methods and methodology: The spectrophotometric method is a common and relatively simple method for determining enzyme activity. This method is based on measuring the ability of an enzyme to convert a substrate into a product. The spectrophotometer is adjusted to the wavelength of maximum absorption of the substrate or product. For example, lactose absorbs most strongly at a wavelength of 280 nm. The "zero" point of the spectrophotometer is adjusted with a buffer. In scientific research, p-nitrophenyl-β-D-galactoside (pNPG) is used as the substrate, the buffer solution is 50 mM sodium phosphate buffer (pH 6.8 or a suitable medium), the stop reagent is 1 M Na₂CO₃ (to stop the reaction), and the standard p-nitrophenol (pNP) solution is used in the spectrophotometer (for measurement at 400 or 405 nm). Results: The methodology used showed that the yield of galactosidase was highest at pH 7.0 and 40°C, resulting in a significant enzymatic activity of 1.5 U/ml. In addition, the enzyme extraction methods yielded up to 50% higher galactosidase levels compared to traditional aqueous methods, demonstrating the effectiveness of the optimized conditions in enhancing enzyme recovery. The best result of the galactosidase enzyme isolated from chickpea was recorded at pH=7.0 and t°C=40°C. Result: 1.5 U/ml. Conclusion: To conclusion, it can be said that as a result of the galactosidase enzyme in the extract prepared from chickpea (Cicer arietinum) seeds has been proven. Analyses conducted on the basis of the ONPG substrate showed that the enzyme works with high activity. The active functioning of this enzyme indicates its promising use in bioindustry and food biotechnology.