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Vacuole and mitochondria patch protein Mcp1 of Saccharomyces boulardii impairs the oxidative stress response of Candida albicans by regulating 2-phenylethanol

Microbial Cell Factories volume 24, Article number: 92 (2025) Cite this article

Abstract

Background

Vacuole and mitochondria patch (vCLAMP) protein Mcp1 is crucial in eukaryotic cells response to environmental stress, but the mechanism of Mcp1 in Saccharomyces boulardii (S. boulardii) against pathogenic fungi is unclear.

Results

This work first explored the role of Mcp1 in S. boulardii against Candida albicans (C. albicans). The results showed that Mcp1 located on the vacuolar and mitochondrial membrane of S. boulardii. Overexpression of Mcp1 inhibited the adhesion and hyphal formation of C. albicans in vitro. The mice model of intestinal infection revealed that WT-pGK1-MCP1 mutant enhanced the ability of S. boulardii antagonize C. albicans infecting gut. High performance liquid chromatography-mass spectrometry analysis demonstrated that overexpressing Mcp1 promoted the production of 2-phenylethanol. The latter is a secondary metabolite of S. boulardii, and can inhibit the adhesion and biofilm formation of C. albicans. The reverse transcription polymerase chain reaction and western blotting results confirmed Mcp1 promoted the production of 2-phenylethanol by regulating the expression level of Aro10. Notably, RNA-sequencing and Gene Ontology enrichment analyses showed that 2-phenylethanol impaired the oxidative stress response of C. albicans.

Conclusion

This work reveals the critical role of Mcp1 in S. boulardii against C. albicans by regulating 2-phenylethanol metabolism, which provide a theoretical basis for S. boulardii as antifungal biologic therapy to prevent and treat of Candida infection.

Background

Candida albicans (C. albicans), a conditionally pathogenic fungus in clinic [1], can cause host systemic infections and even mortality in immunocompromised individuals by invading submucosal tissues of the intestine [2]. The adhesion and hyphal development of C. albicans play a significant role in host invasion [3]. Hitherto, drug therapy has been the classic prevention and treatment strategy for Candida infections. However, the limited antifungal drugs and the emergence of drug-resistant strains bring the great challenge to the clinical treatment of Candida infection [4, 5]. Many studies and our previous work found that the probiotic fungi-Saccharomyces boulardii (S. boulardii) can effectively inhibit virulence of C. albicans [6,7,8], and is considered as a safe and effective alternative therapy for Candida infections.

Recently, membrane contact site, commonly found in mammalian and yeast cells, has caused wide concern owing to their important roles in cell growth and response to environmental stresses [9, 10]. Our previous work found that vacuole and mitochondria patch (vCLAMP) proteins possess the ability to maintain mitochondrial functional stability [11,12,13]. Notably, overexpression of the vCLAMP protein Mcp1 in S. boulardii significantly enhanced the ability of probiotic fungi against C. albicans [6], however, the detailed mechanism has not been elucidated.

Herein, the location of vCLAMP protein Mcp1 was observed by fluorescent labeling and staining techniques. To study the functions of Mcp1 in S. boulardii against C. albicans, firstly, the adhesion and hyphal formation of C. albicans on polystyrene microplate were detected. Secondly, a mice gut infection model was conducted to validate the function of Mcp1 in S. boulardii against C. albicans infecting host intestines. Subsequently, the secondary metabolites of S. boulardii were analyzed using high performance liquid chromatography-mass spectrometry (HPLC-MS). Moreover, the reverse transcription polymerase chain reaction (RT-PCR) and western blotting were used to analyze 2-phenylethanol related gene and protein expression levels. Furthermore, RNA-sequencing (RNA-Seq) and Gene Ontology (GO) enrichment analyses were carried out to study the mechanism of 2-phenylethanol inhibiting the adhesion and biofilm formation of C. albicans.

Method

Construction of strains

S. boulardii, C. albicans, and plasmids mentioned in this work were listed in Table S1. S. boulardii (ura3::ura3) was derived from Shandong Agricultural University (Shandong Province, China). The mutants were constructed using S. boulardii (ura3::ura3) as the parental strain. WT-pGK1-MCP1 mutant was obtained by transferring pSP-G1-MCP1-URA3 plasmid into S. boulardii [14]. Amplifying MCP1-URA3 gene knockout cassette using the primers of MCP1-5DR and MCP1-3DR, and then transformed it into S. boulardii to construct mcp1Δ/Δ mutant strain (Figure S1). The relevant primer sequences were listed in Table S2.

Microscopic analyses

WT-pGK1-MCP1-GFP cells were incubated in YPD medium for 12 h. Adjusting the density of cells (OD600 nm = 0.1) with SC-Uri liquid medium, and then incubating cells at 30 °C for 4 h. The collected cells were stained by MitoTracker Red (dissolved in dimethyl sulfoxide, 1 mM) and observed by a fluorescence microscope. In addition, the collected cells were stained by N-(3-triethylammoniumpropyl)-4-(6-(4- (diethylamino)phenyl)hexatrienyl)pyridinium dibromide (FM4-64) (50 µg/mL, Sigma, St. Louis, MO, USA) to confirm the location of mcp1-GFP [15].

RNA extraction

C. albicans were cultured at 30 °C for 12 h with 160 rpm, and then transferred into YPD medium and cultured continuously for 4 h. Adjusting to an OD600 of 0.2 in RPMI-1640 medium (RP MI-1640 powder 1.04%, MOPS 0.418%, 80 µg/mL uridine, pH 7.0). After adding 2-phenylethanol with final concentration of 0.02 mg/mL, C. albicans were cultured for 4 h at 37 °C. RNA was extracted via Trizol method according to the previously described method [16], and reverse transcribed into single-stranded complementary DNA (cDNA) by using M-MLV reverse transcriptase. This diluted cDNA was used to the following studies.

Filamentous growth assays

C. albicans were cultured for 10 h and transferred it into YPD medium (80 µg/mL uridine) for 4 h to log phase [17]. The cells were collected by centrifugation (12000 rpm) and washed three times by PBS buffer, and suspended in RPMI-1640 medium (80 µg/mL uridine). Then, C. albicans were incubated with 2-phenylethanol (0.002, 0.02 and 0.2 mg/mL, respectively) at 37 °C for 4 h, and centrifuged at 12,000 rpm for 10 min to collect cells. The obtained cells were stained with fluorescent dye (calcofluor white, CFW) at a concentration of 10 mg/mL for 5 min at 30 °C, and observed using a fluorescence microscope (Leica DM2500).

Biofilm formation of C. albicans

The activated C. albicans were cultured in YPD medium (80 µg/mL uridine) for 4 h at 30 °C. Adjusting to an OD600 of 0.1 in RPMI-1640 medium, and transferring the cells into polystyrene microplate. The samples were cultured at 37 °C for 24 h, and stained by crystal violet to assay the biofilm formation of C. albicans by multi-functional enzyme-linked immunosorbent assay reader (TECAN).

ROS assay

2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA) was used to detect the intracellular ROS level in C. albicans [18]. After cultured at 30 °C for 6 h with 160 rpm, C. albicans were transferred into YPD medium containing 2-phenylethanol with final concentration of 0.02 mg/mL at 30 °C for 4 h. The cells were washed by PBS buffer and collected at 12,000 rpm for 2 min, and then stained with DCFH-DA dye (20 µg/mL) for 30 min at 30 °C. Finally, C. albicans were assayed by multi-functional enzyme-linked immunosorbent assay reader (Ex = 488 nm, Em = 520 nm).

Determination of intracellular ATP and cell activity

The cells cultured in YPD medium were collected for protein extraction. The intracellular ATP content in cells was detected using ATP kits. The cells cultured for 4 h was collected and adjusted to OD600 nm = 0.5. MTT reagent was added to catalyze the formation of crystalline formazan from succinate dehydrogenase inside mitochondria. The latter was dissolved by dimethyl sulfoxide, and detected the absorbance of 570 nm (OD570 nm) by a multifunctional enzyme-linked immunosorbent assay reader.

Immunoblotting

The cells cultured in YPD medium for 12 h were collected to extract protein. A sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting were performed to analyze the cells extracts. The luminescence intensity of substrate was boosted using the specific antibody, anti-Rabbit IgG (PROMEGA W4018), anti-Tubulin (MBL PM054), and anti-GFP (MBL598). The protein imprint was imaged by a western blotting exposure device (Tanon, 4800) [17].

HPLC-MS

The supernatant from cultures of yeasts cells was collected by centrifugation, and filtered with a Millipore 0.22 μm filter. Then, the samples were analyzed by HPLC-MS (Thermo Scientific) using a C18 column.

Adhesion assay of C. albicans

To investigate the antagonistic effect of S. boulardii on C. albicans, polystyrene microplate was used to observe the adhesion of C. albicans. Using RPMI-1640 medium (pH 7.0) as culture medium, a 1:1 mixture of S. boulardii and C. albicans was inoculated into polystyrene well plate, and cultured at 37 °C. The unattached C. albicans was washed away by PBS buffer. The adhesion of C. albicans was determined using crystal violet staining method.

Virulence assay

The mice were placed at a constant temperature of 24 °C, and subjected to a 12 h light/dark cycle. They were fed and drank freely at the Experimental Animal Center of Shandong First Medical University. We tried our best to reduce the suffering and the number of animals in experiments.

Four-week-old C57 BL/6 female SPF mice (Jinan Pengyue Experimental Animal Breeding Co., Ltd) were randomly divided into several groups with 10 rats each. The C. albicans (BWP17a) infection was developed according to previous report with some modifications [19]. Briefly, Penicillin (1.5 mg/mL) and Streptomycin (2 mg/mL) in drinking water was given to mice beginning 3 days before infection, and continued in the entire experiment. Gastric infusion was performed to inoculate C. albicans (a cell suspension of 1 × 108 cells/mL). After the second day of C. albicans infection in mice, 200 uL of S. boulardii (WT, mcp1Δ/Δ, and WT-pGK1-MCP1) were continuously conveyed to the stomach for 7 d, with a cell concentration of 1 × 108 cells/mL. Eight days after Candida infection, the colon in mice were taken and fixed overnight in 4% formalin for observation of sections stained by hematoxylin and eosin.

Results

Deletion of Mcp1 damaged the mitochondrial morphology and functions of S. boulardii

In our previous reported, the vacuole and mitochondria patch (vCLAMP) proteins locate at the membrane junction of the vacuole and mitochondrion. Herein, green fluorescent labelled-Mcp1 (Mcp1-GFP) was constructed to observe the intracellular location of Mcp1. The fluorescent results suggested that Mcp1-GFP was localized on the FM4-64 labelled vacuolar membrane, as well as localized on the MitoTracker Red labelled mitochondrial membrane (Fig. 1A). In normal cells, mitochondria were in a filamentous shape. Deletion of Mcp1 caused the mitochondrial morphology changing from linear to fragmented (Fig. 1B). Moreover, ATP and MTT results further showed that Mcp1 deficiency led to mitochondrial dysfunction in S. boulardii (Fig. 1C and D). In addition, to evaluate the damage degree of mitochondrion, the expression level of mitochondrial DNA (mtDNA) encoding genes (COX2, CIM1, and ATP6), an important indicator for mitochondrial function, were detected [20]. As shown in Figure S2, deletion of MCP1 gene decreased the mtDNA quantity of S. boulardii.

Fig. 1
figure 1

Delection of MCP1 damaged mitochondrial functions of S. boulardii. (A) Mcp1-GFP both located on vacuolar and mitochondrial membrane. After cultured in SC-Uri, the cells were collected and stained by Mito Tracker Red and FM4-64 and observed by fluorescence microscope. Bar = 5 μm. (B) The morphology of mitochondria in S. boulardii (WT, mcp1Δ/Δ, and WT + pGK1-MCP1). The samples were stianed by Mito Tracker Red. Bar = 5 μm. Deletion of MCP1 decreased (C) cellar activity and (D) ATP production of S. boulardii. After the stains were mixed with MTT at 30℃ for 1 h, then the samples were resuspended in DMSO, and measured at OD570nm by Multi functional enzyme-linked immunosorbent assay reader. The intracellular ATP contents were detected using an ATP kit. This value represents the mean ± SD from three replicates. Statistical significance was defined based on the different p-values: *p < 0.05, **p < 0.01, and ***p < 0.001, ns: The comparison between the two sets of data was not statistically significant

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Mcp1 is essential in S. boulardii against C. albicans

Recent studies found that probiotic fungi can inhibit the invasion of C. albicans. Herein, overexpression of Mcp1 of S. boulardii effectively reduced the adhesion and biofilm formation ability of C. albicans in vitro (Fig. 2A and B). To demonstrate the biosafety of S. boulardii used in vivo, an oral-gavage method was applied to deliver S. boulardii to mice. As shown in Fig. 2C, the heart, liver, lung, and kidney tissues of mice after oral-gavage treatment were similar to that of normal groups, indicating that S. boulardii is harmless to the body.

Subsequently, a mice gut infection model was conducted to validate the function of Mcp1 in S. boulardii against C. albicans. The mice colon was stained using hematoxylin and eosin staining (H&E) staining method. HE staining results showed that overexpression of Mcp1 in S. boulardii significantly reduced the intestinal invasion ability of C. albicans (Fig. 2D). These results demonstrated that Mcp1 of S. boulardii plays an important role in the antagonism process of C. albicans infecting host.

Fig. 2
figure 2

Overexpression of Mcp1 of S. boulardii inhibited the (A) adhesion and (B) biofilm formation of C. albicans. (C) The heart, liver, lung, and kidney tissues of mice by H&E staining after oral-gavage treatment. Bar = 50 μm. (D) Overexpression of Mcp1 reduced the ability of C. albicans infecting the host gut. Bar = 50 μm. This value represents the mean ± SD from three replicates. Statistical significance was defined based on the different p-values: *p < 0.05, **p < 0.01, and ***p < 0.001, ns: The comparison between the two sets of data was not statistically significant

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Mcp1 plays an important role in S. boulardii against C. albicans by regulating 2-phenylethanol

As reported, the secondary metabolites of probiotics play a major role in the antagonism of pathogenic microorganisms. In present work, we found that the secondary metabolites of S. boulardii inhibited the adhesion and biofilms formation of C. albicans (Fig. 3A, B, and C). After co-cultured with C. albicans, the secondary metabolites of S. boulardii were analyzed using HPLC-MS (Figs. 3D and S3). The results suggested that overexpression of Mcp1 significantly increased the production of 2-phenylethanol, and deletion of Mcp1 declined the production of 2-phenylethanol of S. boulardii (Fig. 3E).

Further studies found that 2-phenylethanol effectively inhibited the adhesion and hyphal formation of C. albicans (Fig. 3F and G). Notably, the hyphal length of C. albicans in the treated and untreated groups were analyzed by Image J. As shown in Figure S4, the hyphal length of treated samples gradually decreased as the concentration of 2-phenylethanol increased from 0.002 to 0.2 mg/mL. The hyphal length of control group was more than five times that of the treated samples which co-cultured with 2-phenylethanol at 0.2 mg/mL. Based on these results, we speculate that Mcp1 of S. boulardii may antagonize C. albicans by regulating 2-phenylethanol metabolism. To verify our hypothesis, RT-PCR and western blotting were used to detect the expression levels of key proteins involved in 2-phenylethanol metabolism. RT-PCR results suggested that overexpression of Mcp1 increased the expression level of ARO10 (Fig. 3H). Many literatures reported that Aro10 is a key catalytic protein in 2-phenylethanol production [21, 22]. Western blotting results further showed that the protein expression level of Aro10 was enhanced by overexpressing Mcp1 (Fig. 3I and J and S5). These results suggested that Mcp1 plays an important role in S. boulardii against C. albicans by regulating 2-phenylethanol.

Fig. 3
figure 3

The secondary metabolites of S. boulardii inhibited the (A and B) adhesion and (C) biofilm formation of C. albicans. (D) The characteristic peak of 2-phenylethanol is 105.0698 (m/z). (E) Characteristic of 2-phenylethanol content in secondary metabolites of S. boulardii. Cells (WT, mcp1Δ/Δ, and WT-pGK1-MCP1) cultured medium were collected, measured and analyzed by HPLC-MS. (F) 2-phenylethanol inhibited hyphal development of C. albicans. After C. albicans cultured in YPD at 30℃, the cells were collected and resuspended in RPMI-1640 medium with 2-phenylethanol (0.002 mg/mL, 0.02 mg/mL, and 0.2 mg/mL) at 37℃ for 4 h. Then, the samples were stained by CFW, and observed by microscope. Bar = 10 μm. (G) 2-phenylethanol reduced the adhesion ability of C. albicans. (H) Deletion of MCP1 decreased the gene expression level of ARO10. The genes related to the metabolism of 2-phenylethanol (ARO1, PHA2, and ARO10) were analyzed by RT-PCR using ACT1 as the normalization genes. (I) Deletion of MCP1 led to abnormal expression of Aro10 in S. boulardii. The cells were collected, and the total protein was extracted. After the samples were detected by western blotting using anti-GFP and anti-tubulin antibodiers. The results were presented through an exposure device. (J) Gray value analysis of Aro10 by Image J. This value represents the mean ± SD from three replicates. Statistical significance was defined based on the different p-values: *p < 0.05, **p < 0.01, and ***p < 0.001, ns: The comparison between the two sets of data was not statistically significant

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Mcp1 of S. boulardii damage the oxidative stress response of C. albicans by regulating 2-phenylethanol

To investigate the mechanism of Mcp1 inhibited the adhesion and biofilm formation of C. albicans by regulating 2-phenylethanol, the intracellular RNA expression levels of C. albicans after 2-phenylethanol treatment were analyzed using RNA-sequencing (RNA-Seq). Transcriptomic analysis showed that 3162 genes (1458 upregulated and 1704 downregulated) were differentially expressed, suggesting that 2-phenylethanol treatment caused a substantial difference in gene expression of C. albicans (Fig. 4A). Gene Ontology (GO) enrichment analyses further suggested that 2-phenylethanol led to abnormalities in oxidative stress response of C. albicans (Fig. 4B). The expression level of genes related to oxidative stress response, such as TRR1, SOD1 and CAT1 were analyzed by RT-PCR. As shown in Figs. 2 and 4C-phenylethanol treatment resulted in an obvious decrease in the expression levels of TRR1, SOD1 and CAT1. These results suggested that 2-phenylethanol may damage the oxidative response process of C. albicans.

Fig. 4
figure 4

(A) Transcriptomic analysis of differential gene expression of C. albicans. After the C. albicans was cultured with 2-phenylethanol, the genes expression of C. albicans were detected by RNA-seq. (B) GO enrichment analyses of differential gene expression. (C) After cultured with 2-phenylethanol, the expression level of genes related to oxidative stress response in C. albicans, including TRR1, SOD1 and CAT1 were assayed by RT-PCR. (D) The expression level of hypha-associated genes including ALS1, ALS3 and HWP1 in C. albicans after cultured with 2-phenylethanol. (E) ROS level, (F) SOD activity, and (G) CAT activity in C. albicans after cultured with 2-phenylethanol. (H) The impact on intracellular ROS levels by deleting and overexpressing CAT1 in C. albicans. This value represents the mean ± SD from three replicates. Statistical significance was defined based on the different p-values: *p < 0.05, **p < 0.01, and ***p < 0.001, ns: The comparison between the two sets of data was not statistically significant

Full size image

Moreover, the expression level of hypha-associated genes including ALS1, ALS3 and HWP1 in C. albicans were also detected by RT-PCR. As expected, the addition of 2-phenylethanol resulted in a significant decrease in the expression levels of the aforementioned hypha-associated genes (Fig. 4D). Furthermore, 2’, 7’-dichlorodihydrofluorescein diacetate (DCFH-DA) staining revealed that the C. albicans exhibited higher intracellular reactive oxygen species (ROS) after treated by 2-phenylethanol (Fig. 4E). In addition, the determination of antioxidant enzyme activity showed that the activity of superoxide dismutases (SOD) and catalase (CAT) declined obviously (Fig. 4F and G). To verify that 2-phenylethanol generated the oxidative stress in C. albicans, the cat1Δ/Δ and pACT1-CAT1 strains were constructed. Compared with WT, the ROS level in cat1Δ/Δ increased, but overexpressing CAT1 decreased the ROS in C. albicans. Moreover, the addition of 2-phenylethanol intensified the level of intracellular ROS in cat1Δ/Δ (Fig. 4H). These results implied that 2-phenylethanol regulated by Mcp1 of S. boulardii damaged the oxidative stress response of C. albicans.

Discussion

C. albicans, a common conditionally pathogenic fungus in clinical practice, easily cause host systemic infections and even endanger life by invading submucosal tissues [23]. The adhesion and hyphal development capabilities of C. albicans play important roles in the process of invading the host [24]. Nowadays, the treatment of Candida infection mainly relies on medication. However, the limited clinical antifungals and the emergence of drug-resistant C. albicans highlight the challenges in the antifungal treatments [25].

S. boulardii, a probiotic fungi isolated from fruits, can effectively inhibit the colonization of C albicans in host intestine [8]. As reported, the vCLAMP mediated by Mcp1 is a novel organelle membrane junction structure. It facilitates information exchange between organelles, and effectively restores the mitochondrial damage caused by ERMES deficiency. Our previous work constructed a S. boulardii bio-coating based on microbial confined growth technology [6]. Interestingly, we found that overexpression of vCLAMP protein Mcp1 of S. boulardii in bio-coating diminished the adhesion and biofilm formation abilities of C. albicans. Overexpression of Mcp1 enhanced antagonistic ability of S. boulardii against the process of C. albicans infecting mice intestine. These studies suggested that Mcp1 plays an important role in S. boulardii against C. albicans, but the detailed mechanism is unclear.

Latest studies found that the secondary metabolites of probiotics could inhabit the growth and reproduction of pathogenic microorganisms [26]. To clarify the mechanism of Mcp1 in S. boulardii against C. albicans, the secondary metabolites of S. boulardii (WT, mcp1Δ/Δ, and WT-pGK1-MCP1) were analyzed using HPLC-MS. The results found that overexpression of Mcp1 significantly enhanced the production of 2-phenylethanol in the secondary metabolites of S. boulardii. Most importantly, 2-phenylethanol reduced the adhesion and hyphal formation of C. albicans.

To validate Mcp1 plays an essential role in S. boulardii against C. albicans by regulating 2-phenylethanol, the expression levels of 2-phenylethanol metabolism related genes (ARO1, PHA2 and ARO10) were detected. RT-PCR results found that overexpression of Mcp1 enhanced the gene expression level of Aro10. As reported, Aro10 is one of the most widely available phenylpyruvate decarboxylase involved in 2-phenylethanol production [21, 22]. Western blotting results further showed that overexpression of Mcp1 enhanced the protein expression level of Aro10 in S. boulardii. These results suggested that Mcp1 may promote the production of 2-phenylethanol by regulating the expression level of Aro10, and then enhanced the ability of S. boulardii against C. albicans.

To further study the mechanism of 2-phenylethanol inhibit the adhesion and biofilm formation of C. albicans, RNA-Seq and GO enrichment analyses were used to analyze RNA expression levels of C. albicans after 2-phenylethanol treatment. The results suggested that 2-phenylethanol may damage the oxidative stress response in C. albicans. RT-PCR results showed that 2-phenylethanol treatment resulted in an obvious decrease in the expression levels of TRR1, SOD1 and CAT1. As well known, antioxidant enzymes (such as SOD and CAT) in organisms can protect cells from oxidative damage by removing ROS. Determination of antioxidant enzyme activity showed that the activity of SOD and CAT significantly reduced. Furthermore, compared with WT, the ROS level in cat1Δ/Δ increased, but overexpressing CAT1 decreased the ROS in C. albicans. Moreover, the addition of 2-phenylethanol intensified the level of intracellular ROS in cat1Δ/Δ. These results indicated that Mcp1 of S. boulardii damaged the oxidative stress response of C. albicans by regulating 2-phenylethanol.

Conclusions

This study revealed that vCLAMP protein Mcp1 promotes the production of 2-phenylethanol by regulating the expression level of Aro10. Interestingly, 2-phenylethanol can damage the oxidative stress response of C. albicans, and then inhibit the adhesion, hyphal formation, and intestinal infection of C. albicans. Therefore, we can infer that Mcp1 of S. boulardii damage the oxidative stress response of C. albicans by regulating 2-phenylethanol. This work enriches the function cognition of organelle membrane junction proteins, and provides new idea for the development of safe and economical antifungal strategy in clinic.

Data availability

No datasets were generated or analysed during the current study.

References

  1. Nishimoto AT, Sharma C, Rogers PD. Molecular and genetic basis of Azole antifungal resistance in the opportunistic pathogenic fungus Candida albicans. J Antimicrob Chemother. 2020;75:257–70.

    Article  CAS  PubMed  Google Scholar 

  2. Ghugari R, Tsao S, Schmidt M, Bonneil É, Brenner C, Verreault A. Mechanisms to reduce the cytotoxicity of Pharmacological nicotinamide concentrations in the pathogenic fungus Candida albicans. FEBS J. 2021;288:3478–506.

    Article  CAS  PubMed  Google Scholar 

  3. Sasaki H, Kurakado S, Matsumoto Y, Yoshino Y, Sugita T, Koyama K, Kinoshita K. Enniatins from a marine-derived fungus Fusarium sp. inhibit biofilm formation by the pathogenic fungus Candida albicans. J Nat Med. 2023;77:455–63.

    Article  CAS  PubMed  Google Scholar 

  4. Lee Y, Puumala E, Robbins N, Cowen LE. Antifungal drug resistance: molecular mechanisms in Candida albicans and beyond. Chem Rev. 2021;121:3390–411.

    Article  CAS  PubMed  Google Scholar 

  5. De Cesare GB, Hafez A, Stead D, Llorens C, Munro CA. Biomarkers of Caspofungin resistance in Candida albicans isolates: a proteomic approach. Virulence. 2022;13:1005–18.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Wei Y, Qiu J, Han Z, Wang X, Zhang H, Hou X, Lv X, Mao X. Antifungal bio-coating of endotracheal tube built by overexpressing the MCP1 gene of Saccharomyces boulardii and employing hydrogel as a house to antagonize Candida albicans. Biomaterials Res. 2023;27:97.

    Article  CAS  Google Scholar 

  7. Kunyeit L, K AA, Rao RP. Application of probiotic yeasts on Candida species associated infection. J Fungi. 2020;6:189.

    Article  CAS  Google Scholar 

  8. Bahuguna N, Venugopal D, Rai N. Biotherapeutic potential of probiotic yeast Saccharomyces boulardii against Candida albicans biofilm. Indian J Microbiol. 2024;10:1090501.

    Google Scholar 

  9. Santos HJ, Nozaki T. Interorganellar communication and membrane contact sites in protozoan parasites. Parasitol Int. 2021;83:102372.

    Article  CAS  PubMed  Google Scholar 

  10. Prinz WA. Bridging the Gap: membrane contact sites in signaling, metabolism, and organelle dynamics. J Cell Biol. 2014;205:759–69.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Mao X, Peng L, Yang L, Zhu M, Du J, Ma C, Yu Q, Li M. Vacuole and mitochondria patch (vCLAMP) and ER-mitochondria encounter structure (ERMES) maintain cell survival by protecting mitochondrial functions in Candida albicans. Biochem Biophys Res Commun. 2022;591:88–94.

    Article  CAS  PubMed  Google Scholar 

  12. Mao X, Yang L, Yu D, Ma T, Ma C, Wang J, Yu Q, Li M. The vacuole and mitochondria patch (vCLAMP) protein Vam6 is crucial for autophagy in Candida albicans. Mycopathologia. 2021;186:477–86.

    Article  CAS  PubMed  Google Scholar 

  13. Mao X, Yang L, Fan Y, Wang J, Cui D, Yu D, Yu Q, Li M. The vacuole and mitochondria patch (vCLAMP) protein Mcp1 is involved in maintenance of mitochondrial function and mitophagy in Candida albicans. Front Microbiol. 2021;12:633380.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Wang T, Sun H, Zhang J, Liu Q, Wang L, Chen P, Wang F, Li H, Xiao Y, Zhao X. The establishment of Saccharomyces boulardii surface display system using a single expression vector. Fungal Genet Biol. 2014;64:1–10.

    Article  PubMed  Google Scholar 

  15. Zhang B, Yu Q, Huo D, Li J, Liang C, Li H, Yi X, Xiao C, Zhang D, Li M. Arf1 regulates the ER-mitochondria encounter structure (ERMES) in a reactive oxygen species-dependent manner. FEBS J. 2018;285:2004–18.

    Article  CAS  PubMed  Google Scholar 

  16. de Souza MR, Teixeira RC, Daúde MM, Augusto ANL, Ságio SA, de Almeida AF, Barreto HG. Comparative assessment of three RNA extraction methods for obtaining high-quality RNA from Candida viswanathii biomass. J Microbiol Methods. 2021;184:106200.

    Article  PubMed  Google Scholar 

  17. Du J, Dong Y, Zhao H, Peng L, Wang Y, Yu Q, Li M. Transcriptional regulation of autophagy, cell wall stress response and pathogenicity by Pho23 in C. albicans. FEBS J. 2023;290:855–71.

    Article  CAS  PubMed  Google Scholar 

  18. Yu D, Zha Y, Zhong Z, Ruan Y, Li Z, Sun L, Hou S. Improved detection of reactive oxygen species by DCFH-DA: new insight into self-amplification of fluorescence signal by light irradiation. Sens Actuators B. 2021;339:129878.

    Article  CAS  Google Scholar 

  19. Takakura N, Sato Y, Ishibashi H, Oshima H, Uchida K, Yamaguchi H, Abe S. A novel murine model of oral candidiasis with local symptoms characteristic of oral thrush. Microbiol Immunol. 2003;47:321–6.

    Article  CAS  PubMed  Google Scholar 

  20. Feng J, Chen Z, Liang W, Wei Z, Ding G. Roles of mitochondrial DNA damage in kidney diseases: A new biomarker. Int J Mol Sci. 2022;23:15166.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Noda S, Mori Y, Ogawa Y, Fujiwara R, Dainin M, Shirai T, Kondo A. Metabolic and enzymatic engineering approach for the production of 2-phenylethanol in engineered Escherichia coli. Bioresour Technol. 2024;406:130927.

    Article  CAS  PubMed  Google Scholar 

  22. Kong S, Pan H, Liu X, Li X, Guo D. De Novo biosynthesis of 2-phenylethanol in engineered Pichia pastoris. Enzym Microb Technol. 2020;133:109459.

    Article  CAS  Google Scholar 

  23. Wagener J, MacCallum DM, Brown GD, Gow NA. Candida albicans Chitin increases arginase-1 activity in human macrophages, with an impact on macrophage antimicrobial functions. mBio. 2017;8:e01820–16.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. McCall AD, Pathirana RU, Prabhakar A, Cullen PJ, Edgerton M. Candida albicans biofilm development is governed by cooperative attachment and adhesion maintenance proteins. NPJ Biofilms Microbiomes. 2019;5:21.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Liang X, Chen D, Wang J, Liao B, Shen J, Ye X, Wang Z, Zhu C, Gou L, Zhou X, et al. Artemisinins inhibit oral candidiasis caused by Candida albicans through the repression on its hyphal development. Int J Oral Sci. 2023;15:40.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Suchodolski J, Derkacz D, Bernat P, Krasowska A. Capric acid secreted by Saccharomyces boulardii influences the susceptibility of Candida albicans to fluconazole and amphotericin B. Sci Rep. 2021;11:6519.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Funding

This study was financially supported by the National Natural Science Foundation of China (82402649), Natural Science Foundation of Shandong Province (ZR2024 MH168, ZR2023QH003 and ZR2022QC117).

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Authors and Affiliations

  1. Shandong First Medical University & Shandong Academy of Medical Sciences, No. 6699 Qingdao Road, Jinan, 250000, Shandong, China

    Yunyun Wei, Chuanqi Li, Jianhao Fu & Xiaolong Mao

  2. School of Radiology, Shandong First Medical University & Shandong Academy of Medical Sciences, Tai’an, China

    Yunyun Wei

  3. School of Laboratory Animal & Shandong Laboratory Animal Center, Shandong First Medical University & Shandong Academy of Medical Sciences, Jinan, Shandong, China

    Wanli Gao & Xiaolong Mao

  4. The Second Affiliated Hospital of Shandong First Medical University& Shandong Academy of Medical Sciences, Tai’an, China

    Xiaohui Zhao

Authors
  1. Yunyun Wei
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  2. Xiaohui Zhao
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  4. Jianhao Fu
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  6. Xiaolong Mao
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Contributions

Y.W. and X.Z. conducted the experiments. C.L., J.F. and W.G. analyzed the data. Y.W. and X.M. prepared the manuscript draft. Y.W. and X.M. designed the whole project.

Corresponding author

Correspondence to Xiaolong Mao.

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Ethics approval and consent to participate

The study was approved by the Ethics Committee of Shandong First Medical University & Shandong Academy of Medical Sciences (permit number W202302270104).

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Informed consent was obtained from all individual participants included in the study.

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The authors declare no competing interests.

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Wei, Y., Zhao, X., Li, C. et al. Vacuole and mitochondria patch protein Mcp1 of Saccharomyces boulardii impairs the oxidative stress response of Candida albicans by regulating 2-phenylethanol. Microb Cell Fact 24, 92 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12934-025-02721-0

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  • DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12934-025-02721-0

Keywords

Microbial Cell Factories

ISSN: 1475-2859

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