Abstract
Background:
Candida albicans is an opportunistic pathogen that can form biofilms, thereby increasing resistance to antifungals such as fluconazole. The extracellular matrix of biofilms rich in β-1,3-glucans is the main barrier to the penetration of antifungal drugs.

Aim:
This study aimed to study the mechanism of recombinant β-1,3-glucanase activity as an antibiofilm agent to increase the effectiveness of antifungals in treating vulvovaginitis candidiasis (VVC).

Methods:
Recombinant β-1,3-glucanase was produced using recombinant DNA technology and tested for its antibiofilm activity against C. albicans biofilms. Tests were carried out at the adhesion stage, biofilm formation, and mature biofilm using the crystal violet assay, scanning electron microscope (SEM), confocal laser scanning microscope (CLSM) methods, and cell viability tests. The combination of β-1,3-glucanase with fluconazole was also evaluated to measure its synergistic effect on biofilm matrix degradation and cell viability.

Results:
The results showed that recombinant β-1,3-glucanase affected all stages of C. albicans biofilm formation. Recombinant β-1,3-glucanase was able to inhibit biofilm matrix formation by 86.05% and reduce C. albicans cell viability by 75.43% when combined with fluconazole. SEM and CLSM analyses revealed significant damage to the 3D structure of the biofilm, demonstrating the synergistic effectiveness of the combination of the two agents. The combination of recombinant β-1,3-glucanase with fluconazole in vitro was shown to be effective in inhibiting biofilm formation and eradicating mature C. albicans biofilms.

Conclusion:
These findings indicate the great potential of β-1,3-glucanase to be applied as an antifungal concomitant agent for overcoming antimicrobial resistance in biofilm infections such as VVC. Further studies are needed to evaluate the effectiveness of the drug in in vivo models and clinical applications.

Key words: Antimicrobial resistance, C. albicans biofilm, Fluconazole, Recombinant β-1,3-glucanase, Vulvovaginitis candidiasis