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Bio-energetics Investigation of Candida albicans Using Real-time Extracellular Flux Analysis
Published on: March 19, 2019
Physiological and transcriptome analysis of Candida albicans in response to X33 antimicrobial oligopeptide treatment
Qunlin Lu1,2,3, Yuanxiu Wang1,2,3, Xing Liao1,2,3
1College of Bioscience and Bioengineering, Jiangxi Agriculture University, Nanchang, China.
Introduction:
Candida albicans is an opportunistic pathogenic fungus, which frequently causes systemic or local fungal infections in humans. The evolution of its drug-resistant mutants necessitate an urgent development of novel antimicrobial agents.
Results:
Here, we explored the antimicrobial activity and inhibitory mechanisms of X33 antimicrobial oligopeptide (X33 AMOP) against C. albicans. The oxford cup test results showed that X33 AMOP had strong inhibitory activity against C. albicans, and its MIC and MFC were 0.625 g/L and 2.5 g/L, respectively. Moreover, SEM and TEM showed that X33 AMOP disrupted the integrity of cell membrane. The AKP, ROS, H2O2 and MDA contents increased, while the reducing sugar, soluble protein, and pyruvate contents decreased after the X33 AMOP treatment. This indicated that X33 AMOP could damage the mitochondrial integrity of the cells, thereby disrupting the energy metabolism by inducing oxidative stress in C. albicans. Furthermore, transcriptome analysis showed that X33 AMOP treatment resulted in the differential expression of 1140 genes, among which 532 were up-regulated, and 608 were down-regulated. These DEGs were related to protein, nucleic acid, and carbohydrate metabolism, and their expression changes were consistent with the changes in physiological characteristics. Moreover, we found that X33 AMOP could effectively inhibit the virulence attributes of C. albicans by reducing phospholipase activity and disrupting hypha formation.
Discussion:
These findings provide the first-ever detailed reference for the inhibitory mechanisms of X33 AMOP against C. albicans and suggest that X33 AMOP is a potential drug candidate for treating C. albicans infections.
Insights
X33 antimicrobial oligopeptide (X33 AMOP) effectively inhibits Candida albicans by disrupting cell membranes and energy metabolism. This novel peptide also reduces virulence factors, showing promise as a new antifungal agent.
Area of Science:
- Microbiology
- Mycology
- Biochemistry
Background:
- Candida albicans is a major human opportunistic fungal pathogen.
- Drug-resistant C. albicans strains necessitate novel antimicrobial therapies.
Purpose of the Study:
- To investigate the antimicrobial activity and inhibitory mechanisms of X33 antimicrobial oligopeptide (X33 AMOP) against C. albicans.
Main Methods:
- Oxford cup diffusion assay, Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC) determination.
- Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM) for cell integrity.
- Biochemical assays for cellular damage and oxidative stress markers.
- Transcriptome analysis to identify gene expression changes.
Main Results:
- X33 AMOP demonstrated potent antifungal activity with MIC of 0.625 g/L and MFC of 2.5 g/L.
- Microscopy revealed disruption of C. albicans cell membrane integrity.
- Treatment induced oxidative stress, altered cellular metabolism, and differential gene expression.
- X33 AMOP inhibited phospholipase activity and hyphal formation, reducing virulence.
Conclusions:
- X33 AMOP exhibits significant antimicrobial and anti-virulence properties against C. albicans.
- The findings elucidate the inhibitory mechanisms, including cell membrane damage and metabolic disruption.
- X33 AMOP represents a promising candidate for developing new treatments against C. albicans infections.
