Characterization of a novel lipase bound to the Geobacillus stearothermophilus AH22 membrane

3 Biotech. 2025 Dec;15(12):417. doi: 10.1007/s13205-025-04584-z. Epub 2025 Nov 20.

Abstract

Lipases (EC 3.1.1.3), used in many industrial applications, hydrolyze tri-, di-, and monoglycerides to free fatty acids and glycerol. This study aimed to determine the capacity of Geobacillus stearothermophilus AH22 for membrane-bound lipase production and to identify the optimal medium for lipase production. Following the determination of certain parameters such as pH and temperature, where the produced membrane-bound lipase exhibits the best activity, the kinetic data, as K m and V max, were obtained in the presence of p-NPA (0.03 ± 0.008 mM and 0.7 ± 0.18 U), p-NPB (0.7 ± 0.18 mM and 3.4 ± 0.40 U), p-NPO (0.12 ± 0.010 mM and 5.2 ± 0.62 U), and p-NPL (0.01 ± 0.003 mM and 0.8 ± 0.15 U) substrates. The highest activity of AH22 membrane-bound lipase was observed at pH 9.0 in the presence of substrates, at temperatures ranging from 20 to 40 °C for substrates with low carbon chains fatty acids and from 50 to 60 °C for substrates with long carbon chains. Also, it was observed that the cells dried by lyophilization showed lipase activity after they were pulverized. However, in terms of reuse, it was determined that wet cells were more efficient than powdered cells. It was found that the activity of the AH22 membrane-bound lipase had not changed much in the presence of anions and cations but decreased significantly in the presence of detergents and β-mercaptoethanol. The AH22 membrane-bound lipase can be directly used in other industrial areas where lipase is used, except for the detergent industry, without requiring additional and expensive processing such as purification or immobilization.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-025-04584-z.

Keywords: Geobacillus stearothermophilus; Industrial enzymes; Membrane-bound lipase; Thermophilic bacteria.