Original Research
Frontiers in Industrial Microbiology ·
Volume 4 - 2026 | https://doi.org/10.3389/finmi.2026.1811171
FERMENZA ®: a patented natural alternative to ketoconazole and zinc pyrithione for managing dandruff and scalp disorders
- Sudip Ghosh1
- Munna Bhattacharya1*corresponding author
- Research and Development Department, Breww Therapeutics Private Limited, Punjab, India
Abstract
Dandruff and related scalp disorders, often caused by Malassezia furfur, pose significant dermatological challenges. This study evaluates FERMENZA ®, a novel, patented formulation derived from pomegranate and beetroot fermented extracts, as a natural alternative to synthetic agents like ketoconazole and zinc pyrithione. In this study, mixed substrate fermentation was utilized for synergistic effect on bioactivity of natural fruits and vegetable extracts. The pomegranate-beetroot fermented extract demonstrated impressive antifungal efficacy against Malassezia furfur MTCC 1374, with percent inhibition of 96 ± 0.01% and 86.7 ± 0.03% compared to active pharmaceutical ingredients ketoconazole and zinc pyrithione, respectively. The investigational formulation FERMENZA ® derived from pomegranate and beetroot fermented extract exhibited the highest zone of inhibition (ZOI) of 28 ± 0.12 mm, outperforming marketed products A and B lotions by achieving 93 ± 0.04% and 60 ± 0.09% inhibition, respectively. Minimum inhibitory concentration (MIC) studies revealed potent activity of the formulation at 8mg/ml. The MIC 90 value was observed at 20mg/mL achieving 92 ± 0.01% inhibition compared to ketoconazole. A time-kill assay highlighted the superior antifungal potency of FERMENZA ® against M. furfur, significantly reducing fungal growth intensity within 48 hours, starting from 6 hours of incubation. The antifungal activity of FERMENZA ® is likely due to the result of synergistic interactions among multiple bioactive metabolites produced during fermentation such as gallic acid along with other phytochemicals, which are known to disrupt fungal cell structures. Efficacy and sensory evaluation confirmed FERMENZA ® ’s ability to alleviate dandruff, itching, flaking, and scaling within 10 days, with moderate benefits in reducing hairfall. These results underscore FERMENZA ® ’s potential as a comprehensive, natural solution for managing dandruff and scalp disorders, providing a safer alternative to synthetic antifungal agents.
Keywords
1 Introduction
Dandruff, a prevalent scalp condition caused by the overgrowth of Malassezia species, particularly Malassezia furfur, affects individuals worldwide (Rhimi et al., 2020b; Saunte et al., 2020). Current treatments rely heavily on antifungal agents such as ketoconazole, climbazole, zinc pyrithione, and selenium sulphide (Billamboz and Jawhara, 2023). These pharmaceutical ingredients are commonly formulated in shampoos to alleviate dandruff by reducing flakes and oiliness. However, regular use of these products often leads to side effects, including scalp dryness, sensitivity, microbiome disruption, and even antimicrobial resistance (Chen et al., 2015; Schward, 2016). Such drawbacks have driven consumer interest toward natural, holistic scalp care solutions. The global anti-dandruff market reflects this shift, with projections indicating a compound annual growth rate (CAGR) exceeding 5% between 2024 and 2033. Consumers increasingly demand products that align with wellness trends and environmental values, creating opportunities for innovative cosmeceutical formulations derived from natural ingredients (Deb, 2024; Singh et al., 2025).
Natural alternatives, such as herbal extracts and essential oils, have been extensively researched for their anti-dandruff properties (Ayatollahi et al., 2021; Tessema et al., 2024). Plant-based bioactives are prized for their low toxicity and cost- effectiveness but face challenges such as degradation, limited stability, and reduced shelf life due to their high water content (Tessema et al., 2025). Traditional extraction methods, like solvent and hydro extraction, have been used to isolate these bioactives but often pose concerns regarding toxicity, environmental impact, and efficacy (Khwaza and Aderibigbe, 2023; Tessema et al., 2025). To address these issues, eco-friendly extraction techniques, including microwave-assisted and supercritical fluid extraction, have emerged. While these methods are environmentally sustainable, their complexity and high cost limit widespread adoption (Chemat et al., 2017). Most existing natural formulations rely on conventional plant extracts without leveraging fermentation-mediated biotransformation that may enhance bioavailability and generate additional bioactive metabolites.
Fermentation offers a promising alternative for developing cosmeceuticals. This process utilizes microorganisms to break down complex compounds, enhancing the safety, stability, and bioavailability of bioactives (Majchrzak et al., 2022). Fermentation also improves the therapeutic efficacy of natural ingredients and produces new active compounds like organic acids, peptides, and antioxidants, making it a sustainable and cost-effective option (Chaiyana et al., 2022; Pérez-Rivero and López-Gómez, 2023). Fermentation-mediated biotransformation can enhance the bioavailability of phytochemicals and generate additional bioactive metabolites, including phenolic compounds and organic acids that may contribute to antimicrobial activity. In addition to academic research, industry practices increasingly support the use of fermentation-derived bioactives in cosmeceutical formulations, for enhancing the efficacy, stability, and bioavailability of natural ingredients, particularly in skin and scalp care products. Fermented extracts such as yeast-derived filtrates, fermented plant extracts, and probiotic-based actives are widely incorporated into commercial formulations due to their improved penetration and bioactivity (Yin et al., 2024). Reports from the cosmetic industry indicate that fermentation-based ingredients are among the fastest-growing segments in personal care, driven by consumer demand for microbiome-friendly and sustainable solutions, with an estimated CAGR of 8-10% from 2023 to 2030 (The Business Research Company, 2024). These industry trends substantiate the hypothesis that fermentation-mediated biotransformation can significantly enhance the antifungal and therapeutic potential of botanical ingredients for dandruff management. Despite growing interest in fermented cosmeceuticals, very few studies have evaluated fermented botanical extracts targeting hair care segment specifically for anti-dandruff applications targeting Malassezia furfur. Though limited, but there are reports on clinical studies of probiotic shampoos containing Bifidobacterium lactis, fermented botanical extracts such as Lactobacillus plantarum-fermented plant ingredients which showed significant improvements in dandruff reduction, scalp health, and hair growth parameters (Reygagne et al., 2017; de Jesus et al., 2023). However, comparative studies assessing fermented natural formulations against established antifungal agents such as ketoconazole and zinc pyrithione remain limited. Therefore, the development of fermentation-derived botanical formulations with validated antifungal activity represents an important research direction.
To address this gap, our research team developed a fermentation-derived botanical formulation, referred to as FERMENZA® which was evaluated in this study as potential natural alternative to synthetic antifungals for Malessezia related scalp conditions. The novelty of this work lies in the use of mixed fruit and vegetable-derived substrates subjected to microbial fermentation using Saccharomyces cerevisiae to enhance the production of bioactive metabolites with antifungal potential (Fermented cider compositions and method of preparation thereof. Indian patent, 2023). In contrast to conventional plant extract-based anti-dandruff formulations, this study investigates fermentation-driven enhancement of antifungal bioactivity and evaluates its efficacy against Malassezia furfur, a key microorganism associated with dandruff. The study further integrates antifungal susceptibility testing, MIC analysis, time-kill assays, physicochemical characterization, and preliminary consumer use evaluation, providing a comprehensive assessment of the fermented formulation. To further validate its effectiveness, FERMENZA®, the investigational formulation was tested against two pharmaceutical agents (ketoconazole and zinc pyrithione) and two leading anti-dandruff products in the Indian market. This is the first time report intregating fermentation biotechnology, antifungal assays, and consumer-based product evaluation. This innovative research aligns with growing consumer demand for natural, sustainable, and effective scalp care solutions. By offering proven results, FERMENZA® addresses Malassezia- associated skin conditions while meeting the expectations of a rapidly evolving anti-dandruff market.
2 Materials and methods
2.1 Materials
Pomegranate (Punica granatum), Beetroot (Beta vulgaris), Banana (Musa acuminata and Musa balbisiana) and Papaya (Carica papaya) were purchased from a local market in Mohali, Punjab. GRAS (Generally regarded as safe) status excipients such as propylene glycol, glycerine, sodium benzoate, hydroxyethylcellulose were procured from M/s E-Merck, India. Microbial strain Malassezia furfur (MTCC 1374) was procured from Microbial Type Culture Collection (MTCC), Chandigarh. Sabourad Dextrose Agar and broth media was procured from HiMedia Laboratories, India. Ketoconazole and Zinc pyrithione API were purchased respectively from Aarti Drugs Pvt. Ltd. and Kumar Organics Pvt. Ltd., Punjab. Marketed product A (2% Ketokonazole lotion), marketed product B (2% Ketoconazole and 1% Zinc pyrithione lotion) and commercially available yeast (Saccharomyces cerevisiae) powder were purchased from local drug store in Mohali, Punjab, India.
2.2 Preparation of fermented extracts and their formulation
Whole fruits and vegetables were washed thoroughly with clean water to remove surface impurities. Three substrate combinations were prepared:
- S1: pomegranate and beetroot (1:1),
- S2: banana and papaya (1:1),
- S3: pomegranate, beetroot, banana, and papaya (1:1:1:1).
Fermentation of the substrate mixtures was carried out using Saccharomyces cerevisiae under controlled conditions. The fermentation process was conducted at 35 °C, pH 7.0 for 72 hours under aerobic conditions using either a laboratory fermentor or shake flask system. Prior to fermentation, the substrate mixtures were homogenized to ensure uniformity. After completion of fermentation, the mixtures were filtered to obtain fermented extracts which were subsequently used for antifungal evaluation and formulation development (Ghosh and Bhattacharya, 2025).All the experiments were done in triplicates. The resulting fermented extracts were labeled as S1, S2, and S3.
FERMENZA®, a water-based formulation, was developed at Breww Therapeutics Pvt. Ltd. Mohali, Punjab, India, as disclosed in the granted Indian Patent No. 459674 (Ghosh and Bhattacharya, 2023). Formulations containing the fermented extracts were prepared using a consistent excipient composition. Formulations with fermented extracts S1, S2, and S3 were labeled S4, S5, and S6, respectively. Ingredients were weighed accurately, and the aqueous base, comprising water, propylene glycol, and glycerin, was heated to 80-90 °C. Fermented extracts and sodium benzoate were then added and stirred for one hour to ensure uniform mixing. The resulting formulations were cooled and packaged in bottles. These formulations were designed for use as hair care products, including but not limited to hair oils, tonics, and serums.
2.3 Preparation of test microorganisms
Malassezia furfur (MTCC 1374) was cultured on SDA supplemented with 1% (v/v) olive oil, which provided essential lipids for fungal growth. Cultures were incubated at 32 °C for 48 hours. A standardized fungal suspension was prepared in sterile saline, adjusting the optical density to match the 0.5 McFarland standard (approximately 1 × 106 CFU/mL). Sterile SDA plates with 1% olive oil were inoculated with this suspension using a sterile cotton swab. Plates were dried under aseptic conditions for 10 minutes before introducing test samples.
2.4 Antifungal susceptibility assay of fermented extracts
The antifungal activity of the fermented extracts (S1, S2, S3) was evaluated using the agar well diffusion method. This method was chosen for its effectiveness against M. furfur (MTCC 1374). Wells of 6 mm diameter were punched into SDA plates using a sterile cork borer. Each well received 50 µL of the test sample (fermented extracts S1, S2, or S3). Positive controls included 50 µL of standard antifungal agents (2% ketoconazole or zinc pyrithione). Plates were incubated at 32 °C for 48 hours in a humidified chamber. All the studies were done in triplicates. Zones of inhibition (ZOI) were measured using a digital caliper, and the growth inhibition percentage was calculated using the formula:
%Growth Inhibition=Zc−Zt/Zc×100
Where Zc is the ZOI for the control and Zt is the ZOI for the test sample.
2.5 Comparative inhibition of FERMENZA® with marketed products
The inhibitory activity of FERMENZA® formulations (S4, S5, S6) against M. furfur (MTCC 1374) was compared with two marketed products: Product A and Product B. Agar well diffusion was used, as described previously. 50 µL of each formulation and marketed product were introduced into the wells. All the studies were done in triplicates. Growth inhibition percentages were calculated for the formulations relative to the marketed products using the same formula mentioned above. Since control ZOI differs between standards, scaling factors vary accordingly.
2.6 Evaluation of physico-chemical and microbial properties of FERMENZA®
Based on optimal growth inhibition against M. furfur (MTCC 1374), the formulation S4, containing fermented pomegranate and beetroot extracts, was selected for further studies and developed into the final product, FERMENZA® hair tonic (Table 1). Physico-chemical and microbial evaluations were conducted at DN Laboratory, Panchkula, Haryana, India, an FDA-approved and ISO 9001:2015, GLP- certified facility. Using pharmacopoeial and in-house methods, the product was analyzed for pH, specific gravity, polyphenol and gallic acid content, preservative levels, synthetic color, heavy metals, and microbial counts (total aerobic and fungal).
Table 1. Investigational product FERMENZA® as anti-dandruff formulation.
| Formulation S4 composition | Formulation S5 composition | Formulation S6 composition | Quantity % |
|---|---|---|---|
| Fermented extract S1 Pomegranate: Beetroot (1:1) | _ | _ | 50 to 70 |
| _ | Fermented extract S2 Banana: Papaya (1:1) | _ | 50 to 70 |
| _ | _ | Fermented extract S3 Pomegranate: Beetroot: Banana: Papaya (1:1:1:1) | 50 to 70 |
| Propylene Glycol | Propylene Glycol | Propylene Glycol | 5 to 20 |
| Glycerin | Glycerin | Glycerin | 5 to 10 |
| Menthol | Menthol | Menthol | 0.1 to 5 |
| Camphor | Camphor | Camphor | 0.1 to 5 |
| Sodium Benzoate | Sodium Benzoate | Sodium Benzoate | 0.01 to 0.1 |
| Water | Water | Water | 20 to 50 |
2.7 Minimum inhibitory concentration assay
The formulation S4 (pomegranate and beetroot extract) was selected for further development into the final product, FERMENZA®, based on its superior growth inhibition. MIC values against M. furfur (MTCC 1374) were determined using a modified method based on Gonelimali et al (Gonelimali et al., 2018). Serial dilutions of FERMENZA® (2–20 mg/mL) were tested. Ketoconazole (2% w/v in propylene glycol) was used as a positive control, and propylene glycol alone served as a negative control. Wells of 6 mm diameter were loaded with varying concentrations of FERMENZA®, and plates were incubated at 32 °C for 48 hours. All the studies were done in triplicates. The MIC50 and MIC90 values were defined as the lowest concentrations achieving >50% and >90% growth inhibition, respectively.
2.8 Time-kill assay
The time-kill assay assessed the fungistatic or fungicidal effects of FERMENZA® on M. furfur (MTCC 1374). A fungal suspension (1 × 106 CFU/mL) was exposed to FERMENZA® at 2× MIC50 concentrations, Marketed product A and Marketed product B respectively. Fungal suspension without any antifungals was served as control. Samples were collected at intervals (0, 6, 12, 18, 24, 30, 36, and 48 hours), serially diluted, and plated on SDA plates, which were incubated at 32 °C for 48 hours, and colony counts (CFU/mL) were recorded (Table 2). All the studies were done in triplicates. Growth reduction over time was plotted to determine the antifungal kinetics.
Table 2. Time-killing assay findings.
| Time (h) | Control (Log CFU/mL) | FERMENZA® (Log CFU/mL) | Marketed product A (Log CFU/mL) | Marketed product B (Log CFU/mL) |
|---|---|---|---|---|
| 0 | 6.0 ± 0.10 | 6.0 ± 0.10 | 6.0 ± 0.10 | 6.0 ± 0.10 |
| 6 | 6.5 ± 0.15 | 5.5 ± 0.15 | 5.0 ± 0.15 | 4.5 ± 0.20 |
| 12 | 8.5 ± 0.20 | 4.2 ± 0.15 | 4.0 ± 0.15 | 3.5 ± 0.20 |
| 18 | 9.0 ± 0.15 | 3.1 ± 0.20 | 3.0 ± 0.15 | 2.5 ± 0.20 |
| 24 | 9.5 ± 0.15 | 2.3 ± 0.15 | 2.0 ± 0.10 | 1.5 ± 0.15 |
| 30 | 9.7 ± 0.10 | 1.8 ± 0.10 | 1.5 ± 0.10 | 1.0 ± 0.15 |
| 36 | 9.8 ± 0.10 | 1.6 ± 0.10 | 1.5 ± 0.10 | 0.5 ± 0.10 |
| 48 | 9.8 ± 0.10 | 1.5 ± 0.10 | 1.4 ± 0.10 | 0.4 ± 0.10 |
Values represent mean ± standard deviation (SD) from three independent experimental replicates (n = 3). CFU, Colony Forming Units.
2.9 FERMENZA® efficacy assessment
An open-label exploratory consumer use assessment was conducted involving 25 volunteers aged between 25–45 years presenting with moderate to severe dandruff symptoms. Participants were provided with 100 mL of FERMENZA® hair tonic and instructed to apply the product daily for 10 days while avoiding other anti-dandruff treatments. The study was designed as a preliminary observational assessment to evaluate user experience and potential scalp health benefits. Product performance was evaluated using a simple 0–5 observational scoring scale assessing parameters such as dandruff severity, itching, flaking, scaling, and hair fall. Since the consumer use assessment was designed as a small exploratory observational study, formal statistical comparisons were not performed. The study did not include randomization, blinding, placebo control, or comparator treatment groups.
2.10 FERMENZA® sensory and safety assessment
FERMENZA® was evaluated for sensory attributes and safety using an eight-parameter scoring system. Parameters included appearance, odor, stickiness, oiliness, ease of spreading, dryness, allergic reactions, and burning sensation. Each parameter was rated on a 0–5 scale, where [0] indicated no effect and [5] indicated the highest intensity. Sensory assessment focused on product color, consistency, and user experience, while safety evaluation captured adverse reactions such as redness, swelling, or discomfort.
2.11 Statistical analysis
All experiments were performed in triplicate (n = 3) unless otherwise specified. Results are expressed as mean ± standard deviation (SD). Statistical analysis was conducted using one-way analysis of variance (ANOVA) followed by Tukey’s post-hoc test to compare differences among groups. A p-value < 0.05 was considered statistically significant.
3 Results
3.1 Inhibitory activity of fermented extracts against M. furfur (MTCC 1374)
The antifungal potential of the fermented extracts was evaluated using the Zone of Inhibition (ZOI) in comparison to marketed antifungal agents Ketoconazole and Zinc Pyrithione. The results are presented in Table 3. Among the tested samples, Sample S1 (Pomegranate and Beetroot fermented extract) exhibited the highest activity, with ZOI values of 24 ± 0.15 mm against Ketoconazole and 26 ± 0.19 mm against Zinc Pyrithione. Sample S2 (Banana and Papaya fermented extract) demonstrated moderate activity, with ZOI values of 21 ± 0.09 mm and 22 ± 0.14 mm, respectively, whereas Sample S3 (pomegranate, beetroot, banana, and papaya mixture) showed strong but slightly reduced activity compared with S1. Statistical analysis using one-way ANOVA followed by Tukey’s post-hoc test indicated that the antifungal activity of Sample S1 was significantly higher (p < 0.05) than that of Samples S2 and S3. The percent inhibition of Sample S1 was found to be 96 ± 0.01% and 86.7 ± 0.03% relative to Ketoconazole and Zinc Pyrithione, respectively (Figure 1). These results suggest that fermentation of pomegranate and beetroot extracts produces metabolites with enhanced antifungal activity against M. furfur.
Table 3. Fermented extracts as natural active for M.furfur (MTCC 1374) growth inhibition.
| Fermented extract | Zone of inhibition in mm(w.r.t Ketoconazole) | %ZOI inhibition(w.r.t Ketoconazole) | Zone of inhibition in mm(w.r.t Zinc pyrithione) | %ZOI inhibition(w.r.t Zinc pyrithione) |
|---|---|---|---|---|
| S1 | 24 ± 0.15 | 96 ± 0.01 | 26 ± 0.19 | 86.7 ± 0.03 |
| S2 | 21 ± 0.09 | 84 ± 0.04 | 22 ± 0.14 | 73.4 ± 0.05 |
| S3 | 23 ± 0.11 | 92 ± 0.02 | 24 ± 0.07 | 80 ± 0.04 |
Values are presented as mean ± standard deviation (SD) from three independent experiments (n = 3). ZOI, Zone of inhibition; SD, Standard deviation.
3.2 Comparative activity of investigational formulation on M. furfur (MTCC 1374) growth inhibition
The investigational formulations (S4, S5, and S6) exhibited varied antifungal efficacy represented in Table 4. S4 (derived from Pomegranate and Beetroot) displayed the highest ZOI of 28 ± 0.12 mm, achieving a 93 ± 0.04% inhibition compared to Marketed Product A lotion. S5 (Banana and Papaya- based) and S6 (combination of all four extracts) showed ZOIs of 22 ± 0.05 mm and 26 ± 0.15 mm, with 73 ± 0.09% and 86 ± 0.06% inhibition, respectively (Figure 2). When benchmarked against Marketed Product B lotion, S4 again demonstrated superior activity with a ZOI of 27 ± 0.14 mm, followed by S6 (24 ± 0.05 mm) and S5 (21 ± 0.08 mm). Statistical analysis revealed that the inhibitory activity of formulation S4 was significantly greater than that of S5 (p < 0.05) and moderately higher than that of S6. These results further indicate pomegranate–beetroot fermented formulation possesses enhanced antifungal efficacy relative to other tested formulations.
Table 4. Efficacy of investigational formulation vs. marketed products.
| FERMENZA®formulation samples | Zone of inhibition in mm(w.r.t Marketed product A lotion) | %ZOI inhibition(w.r.t Marketed product A lotion) | Zone of inhibition in mm(w.r.t Marketed product B lotion) | %ZOI inhibition(w.r.t Marketed product B lotion) |
|---|---|---|---|---|
| Formulation S4 | 28 ± 0.12 | 93 ± 0.04 | 27 ± 0.14 | 60 ± 0.09 |
| Formulation S5 | 22 ± 0.05 | 73 ± 0.09 | 21 ± 0.08 | 48 ± 0.11 |
| Formulation S6 | 26 ± 0.15 | 86 ± 0.06 | 24 ± 0.05 | 54 ± 0.04 |
Values are expressed as mean ± standard deviation (SD) obtained from triplicate experiments (n = 3).
ZOI, Zone of inhibition; SD, Standard deviation.
3.3 Physico-chemical properties of FERMENZA® hair tonic
The FERMENZA® hair tonic appeared as a transparent, light brownish, viscous liquid with a mild and pleasant fragrance. The pH and specific gravity were measured at 6.7 and 1.14, respectively. High levels of total polyphenols (5.05% w/w) and gallic acid (19.90 ppm) were determined using an HPLC method (in-house). No traces of artificial color or heavy metals (lead, arsenic, mercury, and cadmium) were detected in the tonic, making it a safe and effective choice for hair care with minimal risk of adverse effects. The total viable aerobic count and fungal count were recorded as <10 cfu/ml, which is well within the permissible limits set by official pharmacopoeial standards (Table 5).
Table 5. Physicochemical evaluation of FERMENZA® hair tonic.
| Parameter | Result | Limit |
|---|---|---|
| Description | Pale yellow colour oily liquid filled in plastic bottle | – |
| pH Value | 6.71 | – |
| Specific Gravity | 1.136841 gm/ml | – |
| Polyphenol | 5.05% | – |
| Sodium Benzoate | 0.73 ppm | – |
| Gallic Acid | 19.90 ppm | – |
| Ethyl Alcohol | 0.0078% | – |
| Heavy Metals | ||
| - Lead (as Pb) | <10 ppm (LDL 0.4) | NMT 10 ppm |
| - Arsenic (as As) | Not Detected (LDL 0.4) | NMT 3 ppm |
| - Mercury (as Hg) | Not Detected (LDL 0.4) | NMT 1 ppm |
| - Cadmium (as Cd) | Not Detected (LDL 0.4) | NMT 0.3 ppm |
| Total Parabens | Not Detected | – |
| Microbiological Test | ||
| - Total Viable Aerobic Count | <10 cfu/ml | NMT 10 cfu/ml |
| - Total Fungal Count | <10 cfu/ml | NMT 10 cfu/ml |
| - E. Coli/ml | Absent | Should be absent |
| - Salmonella/ml | Absent | Should be absent |
| - S. Aureus/ml | Absent | Should be absent |
| - P. Aeruginosa/ml | Absent | Should be absent |
All analytical measurements were conducted according to standard laboratory procedures, and microbial analyses were performed in compliance with pharmacopoeial microbiological limits for cosmeticproducts.
LDL, Lower Detectable Limit; NMT, Not More Than; CFU, Colony Forming Units.
3.4 FERMENZA® potency findings from MIC
The Minimum Inhibitory Concentration (MIC) of FERMENZA® demonstrated a concentration- dependent antifungal activity against M. furfur (MTCC 1374). At 8 mg/mL, the MIC threshold was observed with a ZOI of 9.2 ± 0.11 mm (36 ± 0.08% inhibition relative to Ketoconazole) as represented in Table 6. Higher concentrations further increased activity, with the MIC50 value observed at 10 mg/mL (ZOI: 12.7 ± 0.08 mm, 51 ± 0.08% inhibition) and the MIC90 value at 20 mg/mL (ZOI: 23.0 ± 0.02 mm, 92 ± 0.01% inhibition). Statistical analysis confirmed that the increase in antifungal activity with increasing concentration was statistically significant (p < 0.05), supporting the dose-dependent antifungal activity of the formulation.
3.5 FERMENZA® mechanism of action
Time-kill assays (Figure 3) revealed a rapid and sustained reduction in fungal growth by FERMENZA®. Starting at 6 log CFU/mL, fungal growth was significantly reduced to 1.5 log CFU/mL within 48 hours. Statistical analysis indicated that the reduction in fungal growth observed after 24 hours and 48 hours was statistically significant (p < 0.05) compared with the initial fungal load, demonstrating sustained antifungal activity of the formulation. Compared to Marketed Products A and B, FERMENZA® consistently outperformed the former while showing comparable efficacy to the later. Minor variations in results reflect expected biological and experimental variability.
3.6 Efficacy of FERMENZA® in addressing dandruff and scalp concerns
The exploratory consumer use assessment (Figure 4) study of FERMENZA® demonstrated significant improvements in dandruff and related scalp issues following daily application of FERMENZA® for 10 days. Dandruff severity scores decreased from 5 (Day 0) to 0.2 (Day 10), reflecting a 96% reduction. Itching severity dropped from 4.5 to 0.4 (91% improvement), while flaking and scaling were reduced by 85% and 77%, respectively. Hairfall showed a moderate improvement, decreasing by 50% over the study period. Although these observations suggest beneficial effects of the formulation on scalp condition, the results should be interpreted as preliminary exploratory findings, as the study was conducted without randomization, blinding, or placebo control.
3.7 Sensory and safety profile of FERMENZA®
FERMENZA® scored highly in sensory evaluations (Figure 5), with excellent ratings for appearance (5/5) and spreadability (4.5/5). Minimal stickiness (2/5) and oiliness (3/5) were noted, while dryness and burning sensation were negligible (scores of 0.5/5). No allergic reactions were reported, confirming its safety for regular use. These results affirm FERMENZA®’s suitability as a user-friendly product.
4 Discussion
Fermentation technology has gained increasing interest in cosmetic science due to its ability to enhance bioavailability of plant metabolites and produce microbiome-friendly bioactive ingredients. In addition, fermentation processes are generally considered environmentally sustainable, aligning with the growing demand for green biotechnology in cosmetic and dermatological product development. The results of this study demonstrate the potent antifungal activity of fermented botanical extracts, particularly those derived from Pomegranate and Beetroot. Sample S1’s superior ZOI values and percent inhibition underscore the therapeutic potential of fermentation-derived bioactives, which enhance the bioavailability of polyphenols and other antifungal compounds. This highlights the therapeutic potential of fermented botanicals as effective antifungal agents, in alignment with studies by Ziemlewska et al (Ziemlewska et al., 2021), which emphasized the enhanced bioactivity of fermentation- derived products. The superior activity observed in Sample S1 could be attributed to its unique mixed substrate composition pomegranate and beetroot, which are potentially antioxidant rich and fermentation, further led to the production of even more bioavailable therapeutically active compounds with enhanced antifungal potency. Li et al (Li et al., 2024). also demonstrated synergistic effect of plant extracts against M. fufur in their recent study.
The comparative analysis of investigational formulations highlights the efficacy of S4, suggesting that targeted substrate selection and fermentation processes can optimize antifungal potency. Notably, S4 derived from pomegranate and beetroot ferment consistently outperformed the other formulations, emphasizing its potential, FERMENZA® thus developed as a robust candidate in antifungal treatment. The minimum inhibitory concentration of FERMENZA® was measured using the produced inhibition zones of minimum 7 mm or larger in the agar well diffusion assay (Taye et al., 2011). The MIC findings reveal the dose-dependent activity of FERMENZA®, with significant inhibition achieved at relatively low concentrations. The dose-dependent antifungal activity of FERMENZA® highlights its potential as a natural alternative to synthetic antifungal agents.
Physicochemical evaluation of FERMENZA® found its high total phenolic content along with gallic acid content as reported in Table 6. The antifungal activity of FERMENZA® may be associated with phenolic compounds produced or enriched during fermentation, including gallic acid. Its antioxidant properties as reported in earlier study (Ghosh and Bhattacharya, 2025) induce oxidative stress, compromising fungal cell walls and membranes. Fermentation processes often generate a complex mixture of bioactive compounds, including phenolic acids, organic acids, and secondary metabolites, which may act synergistically to enhance antimicrobial efficacy. These results align with earlier studies emphasizing the role of bioactive secondary metabolites, such as gallic acid, in disrupting fungal cell walls and inhibiting growth (Nayeem et al., 2016; Rhimi et al., 2020a). The presence of gallic acid and other phenolic compounds in FERMENZA® likely contributes to its effectiveness, corroborating previous research which reported that phenolic compounds can exhibit antifungal activity even at low concentrations, particularly when present within complex phytochemical matrices (Borges et al., 2013; Kahkeshani et al., 2019; Atta et al., 2023). Furthermore, fermentation-mediated biotransformation can enhance the bioavailability and biological activity of phenolic compounds, potentially increasing their functional impact even at relatively low concentrations. Therefore, the precise mechanism of action remains hypothetical and requires further investigation through metabolomic profiling and targeted mechanistic studies.
Table 6. FERMENZA® minimum inhibitory concentration (MIC) assay findings.
| MIC assay parameters | Fermented extract concentration (mg/ml) in FERMENZA® | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| 2 | 4 | 6 | 8 | 10 | 12 | 14 | 16 | 18 | 20 | |
| ZOI (mm) | 3.0 ± 0.14 | 3.7 ± 0.02 | 6.1 ± 0.05 | 9.2 ± 0.11 | 12.7 ± 0.08 | 18.2 ± 0.04 | 20.2 ± 0.15 | 21.0 ± 0.15 | 22.5 ± 0.03 | 23.0 ± 0.02 |
| % ZOI(w.r.t. 2% Ketoconazole API) | 12 ± 0.02 | 14 ± 0.15 | 24 ± 0.04 | 36 ± 0.08 | 51 ± 0.08 | 72 ± 0.14 | 80 ± 0.12 | 84 ± 0.03 | 90 ± 0.04 | 92 ± 0.01 |
Values represent mean ± standard deviation (SD) from three independent experimental replicates (n = 3).
ZOI, Zone of inhibition; MIC, Minimum inhibitory concentration; SD, Standard deviation.
The consumer use assessment of FERMENZA® further validates its potential as a comprehensive solution for scalp health. The observed reductions in dandruff, itching, and flaking are consistent with its antifungal properties, while the moderate improvement in hairfall suggests additional benefits beyond fungal inhibition. Sensory evaluations confirm its user acceptability, with minimal side effects and high satisfaction ratings. However, the consumer use study was conducted as an open-label exploratory assessment without randomization, blinding, or placebo control, involving a relatively small sample size (n = 25) over a short duration of 10 days. Therefore, the reported findings should be interpreted as preliminary observations rather than definitive evidence of therapeutic efficacy. Larger randomized controlled clinical trials are required to confirm the statistical validation of these findings.
The fermented botanical formulation evaluated in this study demonstrated strong antifungal activity against Malassezia furfur, highlighting the potential of fermentation-derived botanical metabolites for scalp care applications. Several commercially available anti-dandruff and scalp-care formulations now incorporate fermented ingredients to target microbial imbalance and improve scalp health (Reygagne et al., 2017; de Jesus et al., 2023; Yin et al., 2024). The findings of the present study align with these industry practices, supporting the translational potential of FERMENZA® as a scalable and market-relevant alternative to synthetic antifungal agents for other Malassezia related skin disorders.
5 Limitations of the study
The limitation of the study was assessed based on the data which mainly focused on in-vitro experiments using a single reference strain of Malassezia furfur MTCC 1374. Since antifungal susceptibility can vary among clinical isolates, future studies should evaluate the formulation against multiple clinical strains of Malassezia species to validate broader antifungal efficacy. Hence, further studies on clinical isolates and larger human samples are needed for long-term clinical data generation for better understanding of its clinical efficacy and safety.
6 Conclusion
The findings suggest that fermentation-derived botanical formulations may represent promising natural candidates for managing dandruff-associated fungal growth. This study highlights FERMENZA® as a groundbreaking natural alternative to synthetic antifungal agents like ketoconazole and zinc pyrithione for managing dandruff and scalp disorders. The formulation showcased superior antifungal efficacy, effectively inhibiting the growth of Malassezia furfur and outperforming widely used commercial products. Its potent activity stems from its unique composition of pomegranate and beetroot fermented extracts, rich in polyphenols and gallic acid, which disrupts fungal cell integrity and metabolic pathways. FERMENZA® also demonstrated remarkable effectiveness in addressing scalp issues such as dandruff, itching, flaking, and scaling, alongside noticeable hairfall reduction. With strong antifungal properties, excellent safety, and high user satisfaction, FERMENZA® offers a holistic and natural solution for scalp health and hair care. This study underscores its potential as a dependable alternative to conventional treatments, meeting the growing demand for safer, sustainable, and more effective therapeutic options in modern dermatological care.
Generative AI statement
The author(s) declared that generative AI was used in the creation of this manuscript. During the preparation of this work the author(s) used ChatGPT (https://openai.com/index/chatgpt/) in order to improve the language quality of the manuscript. After using this tool, the author(s) reviewed and edited the content as needed and take full responsibility for the content of the published article. The authors have seen and approved the manuscript and it has not been published or considered for publication elsewhere.
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Footnotes
Edited by: Romina Rodríguez-Sanoja, National Autonomous University of Mexico, Mexico
Reviewed by: Tegen Dagnew Tessema, University of Rovira i Virgili, Spain
Jocelin Rizo, National Autonomous University of Mexico, Mexico
Statements
- Conflict of interest
SG, MB were employed by company Breww Therapeutics Private Limited.
- Funding
The author(s) declared that financial support was not received for this work and/or its publication.
- Data availability
The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.
- Ethics approval
The product is a cosmetic/non-drug formulation, No invasive procedures were performed, No biological samples were collected, No clinical diagnosis or medical intervention was made. It was a product performance evaluation, not a clinical trial. Risk level was minimal. The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.
- Author contributions
SG: Data curation, Validation, Writing – review & editing. MB: Conceptualization, Methodology, Supervision, Writing – original draft.
- Acknowledgements
The authors would like to acknowledge Technology Business Incubator, IISER Mohali, Punjab, India for incubation support.
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Citation
Ghosh, S. & Bhattacharya, M. (2026). FERMENZA ®: a patented natural alternative to ketoconazole and zinc pyrithione for managing dandruff and scalp disorders. Frontiers in Industrial Microbiology, Volume 4. https://doi.org/
- Received
- Accepted
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- Volume
- 4 - 2026
Copyright
Copyright © 2026 Ghosh and Bhattacharya.
License: https://creativecommons.org/licenses/by/4.0/
*Correspondence: Munna Bhattacharya
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