Functional microbial diversity in arid soils of Uzbekistan: discovery of a biotechnologically valuable Bacillus amyloliquefaciens strain

Authors

  • Barno Alimova Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Ozodakhon Pulatova Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Shokhrukh Sadullaev Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Mirshod Sailiev Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Mubina Makhsumkhanova Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Rustam Rakhmonov Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Alonurkhon Abdurakhimova Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Mukhammadjon Ismoilov Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan
  • Akhmadzhan Makhsumkhanov Institute of Microbiology of the Academy of Sciences of the Republic of Uzbekistan, Tashkent, Uzbekistan

DOI:

https://doi.org/10.5281/zenodo.18208937

Keywords:

Arid soils, microbial diversity, Bacillus amyloliquefaciens, milk-clotting activity, protease, dairy biotechnology

Abstract

In this study, we assessed the functional microbial diversity of arid and anthropogenically impacted soils of Uzbekistan to identify bacterial strains with biotechnologically important enzymatic activities. A total of 20 Bacillus isolates obtained from oil-sludge–contaminated areas and intensively irrigated agricultural fields were screened for their ability to produce proteases with milk-clotting activity (MCA). Preliminary screening on skim-milk agar revealed three isolates exhibiting strong proteolytic activity (hydrolysis zones > 20 mm). Quantitative assays performed in a lactose-based fermentation medium identified isolate 6/4/2 as the most active strain. MALDI-TOF mass spectrometry and 16S rRNA gene sequencing confirmed this isolate as Bacillus amyloliquefaciens UzRSMMT-413. Under optimal cultivation conditions (35–40 °C; pH 7.5), UzRSMMT-413 reached a maximum MCA of 400 U mL⁻¹ after 48–72 h and demonstrated an MCA/protease activity ratio of 5.18, indicating high specificity toward casein with minimal nonspecific proteolysis. These findings highlight the arid soils of Uzbekistan as an ecologically rich yet understudied reservoir of microbial resources with significant biotechnological potential. The functional diversity identified in these soils offers promising enzymatic candidates for cheese making, fermented dairy production, and other low-energy bioprocesses. Overall, the study underscores the conservation value and applied importance of microbial biodiversity within the semi-natural landscapes of the Palearctic–Oriental transition zone.

References

Alahmad Aljammas, H., Yazji, S., & Azizieh, A. (2022). Optimization of protease production from Rhizomucor miehei Rm4 isolate under solid-state fermentation. Journal of Genetic Engineering and Biotechnology, 20, 82. https://doi.org/10.1186/s43141-022-00358-9

Mohsin, A. Z., Norsah, E., Marzlan, A. A., Abd Rahim, M. H., & Meor Hussin, A. S. (2024). Exploring the applications of plant-based coagulants in cheese production: A review. International Dairy Journal, 148, 105792. https://doi.org/10.1016/j.idairyj.2023.105792

Anson, M. L. (1938). The estimation of pepsin, trypsin, papain and cathepsin with hemoglobin. Journal of General Physiology, 22, 79–89.

Arima, K., Iwasaki, S., & Tamura, G. (1967). Milk clotting enzyme from microorganisms: Part I. Screening test and identification of the potent fungus. Agricultural and Biological Chemistry, 31(5), 540–551.

Dobozi, R., Jákói, Z. P., Csanádi, J., & Beszédes, S. (2023). Investigating the acid- and enzyme-induced coagulation of raw milk using dielectric and rheological measurements. Applied Sciences, 13(10), 6185. https://doi.org/10.3390/app13106185

Dutt, K., Gupta, P., Saran, S., Misra, S., & Saxena, R. K. (2009). Production of milk-clotting protease from Bacillus subtilis. Applied Biochemistry and Biotechnology, 158(3), 761–772. https://doi.org/10.1007/s12010-008-8504-9

Karam, E. A., Hassan, M. E., Elattal, N. A., Kansoh, A. L., & Esawy, M. A. (2024). Cell immobilization for enhanced milk-clotting enzyme production from Bacillus amyloliquefaciens and cheese quality. Microbial Cell Factories, 23(1), 283. https://doi.org/10.1186/s12934-024-02521-y

Lemes, A. C., Pavón, Y., Lazzaroni, S., Rozycki, S., Brandelli, A., & Kalil, S. J. (2016). A new milk-clotting enzyme produced by Bacillus sp. P45 applied in cream cheese development. LWT – Food Science and Technology, 66, 217–224. https://doi.org/10.1016/j.lwt.2015.10.038

Liburdi, K., Emiliani Spinelli, S., Benucci, I., Lombardelli, C., & Esti, M. (2018). A preliminary study of continuous milk coagulation using Cynara cardunculus flower extract and calf rennet immobilized on magnetic particles. Food Chemistry, 239, 157–164. https://doi.org/10.1016/j.foodchem.2017.06.093

Lowry, O. H., Rosebrough, N. J., Farr, A. L., & Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry, 193(1), 265–275. https://doi.org/10.1016/S0021-9258(19)52451-6

Meng, F., Chen, R., Zhu, X., Lu, Y., Nie, T., Lu, F., & Lu, Z. (2018). Newly effective milk-clotting enzyme from Bacillus subtilis and its application in cheese making. Journal of Agricultural and Food Chemistry, 66(24), 6162–6169. https://doi.org/10.1021/acs.jafc.8b01697

Naveed, M., Tianying, H., Wang, F., Yin, X., Chan, M. W. H., Ullah, A., & Khan, A. M. (2022). Isolation of lysozyme-producing Bacillus subtilis strains, identification of a new strain Bacillus subtilis BSN314 with the highest enzyme production capacity, and optimization of culture conditions for maximum lysozyme production. Current Research in Biotechnology, 4, 290–301.

Yamazaki, H., Ohnishi, Y., Takeuchi, K., Mori, N., Shiraishi, N., Sakata, Y., Suzuki, H., & Horinouchi, S. (1999). Genetic transformation of a Rhizomucor pusillus mutant defective in asparagine-linked glycosylation: Production of a milk-clotting enzyme in a less-glycosylated form. Applied Microbiology and Biotechnology, 52(3), 401–409. https://doi.org/10.1007/s002530051538

Zhang, Y., Hu, J., Wang, J., Liu, C., Liu, X., Sun, J., Song, X., & Wu, Y. (2023). Purification and characteristics of a novel milk-clotting metalloprotease from Bacillus velezensis DB219. Journal of Dairy Science, 106(10), 6688–6700. https://doi.org/10.3168/jds.2023-23450

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Published

2026-01-13

How to Cite

Alimova, B., Pulatova, O., Sadullaev, S., Sailiev, M., Makhsumkhanova, M. ., Rakhmonov, R. ., Abdurakhimova, A., Ismoilov, M. ., & Makhsumkhanov, A. . (2026). Functional microbial diversity in arid soils of Uzbekistan: discovery of a biotechnologically valuable Bacillus amyloliquefaciens strain. Journal of Wildlife and Biodiversity, 9(4), 123–133. https://doi.org/10.5281/zenodo.18208937