Enzymatic disruption of pathogenic biofilms by thermostable endoglucanase from Bacillus cereus ARA-12: Insights for pharmaceutical and industrial use

This study focused on the partial purification and characterization of endoglucanase produced by Bacillus cereus ARA-12, with the special emphasis on its anti-biofilm potential against pathogenic bacteria. Partial purification was achieved through ammonium sulfate precipitation, resulting in a 6.97 purification fold with a specific activity of 91.519 U/mg. The optimum temperature and pH of the enzyme were found to be 50°C and 8.0 respectively. The enzyme retained over 80% of its residual activity after 30 minutes of incubation at high temperatures. The kinetic studies revealed a Km of 3.25 mg/mL and a Vmax of 2500 ?mol/min, indicating high substrate affinity and catalytic efficiency. The enhanced enzyme activity was observed with metal ions and the enzyme also demonstrated stability in the presence of detergents as well. The partially purified endoglucanase showed significant anti-biofilm potential. Inhibition studies revealed a maximum biofilm reduction of 72% for Pseudomonas aeruginosa after 36 hours of incubation. Eradication studies demonstrated biofilm detachment rates of 87% for Pseudomonas aeruginosa, 72% for Staphylococcus aureus, and 56% for Escherichia coli after 36 hours. Scanning electron microscopy confirmed structural degradation of biofilms. These findings underscore the enzyme's dual potential for industrial biocatalysis and therapeutic applications, particularly in combating biofilm-associated infections.