Transactions of the Institute of Molecular Biology & Biotechnologies, MSE AR, vol. 9, No 1, p. 12-16 (2025) 12 DOI: https://doi.org/10.62088/timbb/9.1.2 Identification and study of extracellular enzymatic activity of Bacillus velezensis isolated from saline soils of the Fergana Valley Mokhira Narmukhamedova * , Tokhir Khusanov, Gulchekhra Kadirova, Zakhro Akhmedova Institute of Microbiology, Academy of Sciences of the Republic of Uzbekistan, 7B A.Kadyri Str., Tashkent, Republic of Uzbekistan * For correspondence: narmuhamedovamohira@gmail.com Received: May 06, 2025; Reviewed: May 23, 2025; Accepted: June 05, 2025 This study focuses on the extracellular enzymatic activity of the dominant bacterial species Bacillus sp. isolated from saline soils of the Fergana region. Among more than 25 isolates studied, isolate No. 2, obtained from the topsoil layer (0–15 cm), and exhibited the highest hydrolytic activity. Notably high activity was observed for pectinase and amylase, with hydrolysis zones ranging from 38 to 45 mm. Moderate activity was shown by xylanase, cellulase, protease, and lipase, with hydrolysis zones between 24 and 34 mm. Based on molecular genetic analysis of the 16S rRNA gene sequence, the highly active dominant strain was identified as Bacillus velezensis TM. Keywords: Soil, isolates, bacteria, identification, Bacillus velezensis, hydrolases, pectinase, amylase, xylanase, protease, cellulase, lipase INTRODUCTION The selection of low-cost enzyme production methods and the mismatch between optimal operating temperatures are limiting factors. Microorganisms such as bacteria, filamentous fungi, and yeasts can be used for the production of hydrolases, such as amylases and cellulases. Under appropriate working conditions, the production of these enzymes, for example, using Bacillus velezensis – can be a cost-effective alternative to traditional enzyme production methods based on submerged or solid-state fermentation. This is because enzyme biosynthesis directly within microorganisms may help reduce overall process costs, as the use of commercial enzymes is associated with significant economic impact (about 15.7% of the total process cost) (Devos et al., 2024). Bacillus species have become common industrial microbial chassis, with approximately 60% of enzyme products on the market derived from Bacillus. Compared to the complexities of molecular-genetic manipulation in fungi and safety concerns related to endotoxins in Escherichia coli, Bacillus expression systems offer several distinct advantages. These include short expression cycles, high protein yields, the non-pathogenic nature of the host, absence of endotoxins in expressed proteins, excellent thermostability, lack of significant codon bias, and an efficient secretion signal peptide and chaperone system (Ye et al., 2024). Recent advances in microbial genomics and plant-microbe interactions have clarified the mechanisms through which B. velezensis contributes to plant health, making it a promising candidate for sustainable agricultural practices (Bach et al., 2025; Kang et al., 2025). Bacillus velezensis is widely found in various fermented products, such as fermented soybean paste (doenjang) and fermented vegetables like kimchi. The use of B.velezensis as a starter culture has increased and has been reported to affect the technological properties of salted seafood during fermentation. It has also been reported that B.velezensis SS360–1, isolated from traditional soy sauce, influences the production of aromatic compounds in vitro (Na et al., 2023; Peng et al., 2025; Revankar & Bagewadi, 2025). In addition, B.velezensis HF-14,109 was isolated from the intestine of healthy carp (Cyprinus carpio L.) and demonstrated tolerance to extreme environmental conditions, such as high temperatures, acidity, and bile salts. It was also sensitive to various antibiotics and capable of secreting amylase, proteases, and other digestive enzymes, as well as xylanase, β-mannanase, and other non-starch polysaccharide-degrading enzymes (Wang et al., 2024; Zhang et al., 2025). The isolated strain Bacillus velezensis NA16, in a submerged fermentation culture of poultry feathers, produced a large number of proteases, thereby overcoming the common obstacle of high substrate costs typically encountered in industrial enzyme production. During a 72-hour cultivation period, NA16 degraded chicken feathers with an efficiency of 99.3% and produced a high