116 © 2025 by the author; licensee Asian Online Journal Publishing Group Agriculture and Food Sciences Research Vol. 12, No. 2, 116-122, 2025 ISSN(E) 2411-6653/ ISSN(P) 2518-0193 DOI: 10.20448/aesr.v12i2.7530 © 2025 by the author; licensee Asian Online Journal Publishing Group Fermentation characteristics of yeasts isolated from apple and kiwi Hiroko Seki Department of Local Produce and Food Sciences, Faculty of Life and Environmental Sciences, University of Yamanashi, Takeda, Kofu, Yamanashi, Japan. Email: hiroko.sk@yamanashi.ac.jp Abstract This study investigated the fermentation capacity of two natural yeast strains, Metschnikowia pulcherrima and Candida apicola, isolated from apples and fermented kiwi syrup, respectively. Natural yeasts are increasingly valued for imparting complexity, unique aroma, and distinctive flavor to alcoholic beverages and baked goods. However, their fermentation performance is often weaker and less stable than that of commercial dry yeast. To evaluate optimal conditions, each strain was cultured on agar and inoculated into liquid media containing yeast extract, peptone, and glucose. Bread dough was then fermented with each strain under varying temperatures, and sugar types and concentrations. Fermentation efficiency was assessed by measuring dough height. Statistical analysis was performed using t-tests and one-way ANOVA on triplicate data following Fisher’s three principles. Results showed that M. pulcherrima performed best at 25 °C with 5.0% glucose, while C. apicola showed optimal fermentation at 35 °C with 5.0% sucrose. These findings indicate that tailoring fermentation conditions to specific natural yeast strains can significantly enhance the efficiency of alcohol and bread production, offering a promising alternative to commercial yeast in both artisanal and industrial applications. Keywords: Bread production, Candida apicola, Fermentation temperature, Fermentation, Metschnikowia pulcherrima, Sugar type. Citation | Seki , H. (2025). Fermentation characteristics of yeasts isolated from apple and kiwi. Agriculture and Food Sciences Research, 12(2), 116–122. 10.20448/aesr.v12i2.7530 History: Received: 21 August 2025 Revised: 16 September 2025 Accepted: 26 September 2025 Published: 10 October 2025 Licensed: This work is licensed under a Creative Commons Attribution 4.0 License Publisher: Asian Online Journal Publishing Group Funding: This study received no specific financial support. Institutional Review Board Statement: Not applicable. Transparency: The author confirms that the manuscript is an honest, accurate, and transparent account of the study; that no vital features of the study have been omitted; and that any discrepancies from the study as planned have been explained. This study followed all ethical practices during writing. Competing Interests: The author declares that there are no conflicts of interests regarding the publication of this paper. Contents 1. Introduction .................................................................................................................................................................................... 117 2. Materials and Methods ................................................................................................................................................................. 117 3. Results .............................................................................................................................................................................................. 118 4. Discussion ........................................................................................................................................................................................ 120 5. Conclusion ....................................................................................................................................................................................... 121 References ............................................................................................................................................................................................ 121 mailto:hiroko.sk@yamanashi.ac.jp https://creativecommons.org/licenses/by/4.0/ https://creativecommons.org/licenses/by/4.0/ https://www.doi.org/10.20448/aesr.v12i2.7530 https://orcid.org/0000-0003-2426-7554 Agriculture and Food Sciences Research, 2025, 12(2): 116-122 117 © 2025 by the author; licensee Asian Online Journal Publishing Group Contribution of this paper to the literature This study contributes to existing literature by investigating the fermentation characteristics of two natural yeasts. It provides new insights into the effects of temperature, sugar type, and sugar concentration on the fermentation capacity of these yeasts, thereby deepening our understanding of bread making and sake brewing. 1. Introduction Alcohol production is primarily carried out through fermentation using Saccharomyces cerevisiae, although other natural yeasts are also commonly utilized. For instance, Torulaspora delbrueckii is used in beer production [1]. Additionally, Metschnikowia pulcherrima is applied in wine production [2], Schizosaccharomyces pombe in rum production, and Kluyveromyces marxianus in the fermentation of whey from cheese production to produce vodka and gin [3]. In recent years, bread production with natural yeast has garnered considerable interest [4]. Natural yeast is a type of fungus that thrives under diverse environmental conditions. Unlike bread made with commercial dry yeast, bread produced using natural yeast has a complex taste, as different yeasts impart unique aromas and flavors [5]. Currently, natural yeast bread made with strains isolated from dried grapes [6], sudachi petals, and fruits and flowers collected in Tokachi Region, Hokkaido, Japan, is commercially available [7]. Moreover, bread has been successfully produced using Lachancea fermentati, Lachancea kluyveri, and Torulaspora species isolated from rose cultivars grown in Fukuyama City, Hiroshima, Japan [8]. Saccharomyces rouxii, Saccharomyces bisporus, and Saccharomyces exigus have been isolated from grape juice, pineapple juice, and rice obtained from local markets in Savar, Bangladesh [9]. In bread production, yeast fermentation generates carbon dioxide, which causes wheat gluten to expand, and differences in yeast fermentation capacity directly affect bread quality. Commercial yeast has a strong fermentation capacity, resulting in a softer texture. In contrast, natural yeast is less stable, ferments slowly, and generally shows weaker activity [10]. Therefore, when producing bread with natural yeast, it is essential to adjust the sugar type and concentration as well as temperature to stabilize fermentation and improve efficiency. For example, bread can be successfully produced with Hanseniaspora meyeri when monosaccharides such as glucose and fructose are provided at fermentation temperatures of 25–28°C [11]. Kluyveromyces delphensis ferments dextrose and sucrose but not fructose or lactose, as demonstrated by sugar-specific fermentation tests [12]. Furthermore, S. cerevisiae has an optimum growth temperature of 25–30°C and a fermentation temperature of 45°C; it can utilize glucose, fructose, and sucrose but not in the presence of maltose and amylose [13]. Its ethanol production capacity is maximized at a sugar concentration of 125 g/L rather than 100 g/L. However, ethanol productivity decreases at concentrations above 150 g/L [14], suggesting that both sugar type and concentration substantially influence the fermentation efficiency of yeast. As different natural yeasts respond differently during fermentation, appropriate conditions should be selected for each yeast to ensure successful bread production. In this study, I focused on Metschnikowia pulcherrima [15], isolated from fermented apple syrup, and Candida apicola [16], isolated from fermented kiwi syrup. Metschnikowia pulcherrima has previously been isolated from the spontaneous fermentation of Tannat and Marselan grape musts in Concordia (Entre Rios, Argentina) [17], Cabernet Sauvignon wine in Ningxia, China [18], and Marastina wine [19]. Candida apicola has been isolated from naturally fermented Alorena green table olives [20], wine yeasts that ferment grapes with high sugar content [21], and cachaca (sugarcane wine) [22, 23]. Here, the effects of fermentation temperature as well as the sugar type and concentration were assessed to determine the optimal fermentation conditions required by each species in bread production. The results of this study may have potential applications in the production of alcoholic beverages. 2. Materials and Methods Metschnikowia pulcherrima (NRBC0863) was purchased from the NITE Biological Resource Center, and C. apicola was isolated from kiwi syrup (accession number LC878464, DNA Data Bank of Japan). To prepare each yeast solution, yeast monocultured on agar medium was inoculated into a liquid medium containing 0.25% yeast extract, 0.50% peptone, and 0.10% glucose (for M. pulcherrima) or 5.0% glucose (for C. apicola). The inoculum was incubated at 30 °C for approximately 1 week. After incubation, the medium was removed using centrifugation (890 × g, 5 min, 25 °C), and the yeast was weighed. Subsequently, the yeast was suspended in a 1% glucose solution to obtain a yeast solution with a final concentration of 0.78%–3.9%. 2.1. Examination of the Optimal Fermentation Temperature for Each Yeast Yeast solution (3.0 g) and strong flour (3.0 g; Tomizawa Shouten Inc., Tokyo, Japan) were mixed in a test tube (1.5 cm φ × 18 cm) and placed in a hot water bath adjusted to 20–35 °C in a low-temperature cooker (Anova; Axia International Co., Ltd., Tokyo, Japan). The increase in dough height was measured every hour for 5 hours and calculated as the increase in dough height per 0.1 g of yeast. 2.2. Investigation of the Optimal Sugar Type for Each Yeast Yeast solution (3.0 g), strong flour (3.0 g), and sugar (0.15 g; glucose, sucrose, starch, fructose, and maltose) were mixed in a test tube (1.5 cm φ × 18 cm) and placed in a hot water bath at 25°C (M. pulcherrima) and 35°C (C. apicola) for 12 hours, and the fermentation capacity was calculated as the increase in dough height per 0.1 g of yeast. 2.3. Examination of the Effect of Sugar Concentration Yeast solution (3.0 g), strong flour (3.0 g), and sugar (0.03–0.15 g) were mixed in a test tube (1.5 φ × 18 cm) and placed in a hot water bath at 25°C (M. pulcherrima) and 35°C (C. apicola) for 12 hours. The fermentation capacity was calculated as the increase in dough height per 0.1 g of yeast. 2.4. Statistical Analysis Data were obtained in triplicate using the same sample based on Fisher's three principles. The mean differences between groups were assessed using t-tests, and those among three groups were assessed using a one-way analysis Agriculture and Food Sciences Research, 2025, 12(2): 116-122 118 © 2025 by the author; licensee Asian Online Journal Publishing Group of variance. All statistical analyses were performed using Microsoft Excel (Microsoft Corporation, Redmond, WA). The significance level was set at p < 0.05. 3. Results 3.1. Examination of the Optimal Fermentation Temperature for Each Yeast Figure 1 shows the effect of different temperatures on M. pulcherrima fermentation. At 35°C, no change was observed in dough height after 1 hour; however, the dough height increased to 1.3 cm after 5 hours (p < 0.05). At 30°C, the height increased to 1.6 cm after 5 hours (p < 0.05). At 25°C, no change was observed in dough height after 2 hours; however, the dough height increased to 1.7 cm after 5 hours (p < 0.05). At 20°C, no change was observed in dough height after 1 hour, but the height slightly increased to 0.16 cm after 2 hours (p > 0.05). Thereafter, no changes were observed up to 5 hours. Figure 1. Effect of temperature on Metschnikowia pulcherrima fermentation. ● Indicates 35°C, ■ indicates 30°C, ◆ indicates 25°C, and ▲ indicates 20°C. Note: The fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as an integrated value. Measurements were obtained in triplicate. The error bars indicate the standard deviation. Figure 2 shows the effect of temperature on C. apicola fermentation. At 35°C, no change was observed in dough height after 1 hour; however, the dough height increased to 2.0 cm after 5 hours (p < 0.05). At 30°C, the height increased to 1.9 cm after 5 hours (p < 0.05). At 25°C, no change was observed in dough height up to 4 hours, and the height slightly increased to 0.49 cm after 5 hours (p > 0.05). At 20°C, no change was observed up to 2 hours; nevertheless, the height slightly increased to 0.25 cm after 3 hours (p > 0.05) and remained unchanged for up to 5 hours (p > 0.05). Figure 2. Effect of temperature on Candida apicola fermentation. ● Indicates 35 °C, ■ Indicates 30 °C, ◆ Indicates 25 °C, and ▲ Indicates 20 °C. Note: The fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. Measurements were obtained in triplicate. The error bars indicate the standard deviation. Agriculture and Food Sciences Research, 2025, 12(2): 116-122 119 © 2025 by the author; licensee Asian Online Journal Publishing Group 3.2. Investigation of the Optimal Sugar Type for Each Yeast Figure 3 shows the effects of various sugars on M. pulcherrima fermentation. The addition of glucose increased the dough height to 8.1 cm after 12 h (p < 0.05). The addition of sucrose did not alter the dough height until 3 h, but it increased to 0.17 cm after 4 h (p < 0.05). Thereafter, no change was observed in dough height until 6 h (p > 0.05), followed by an increase to 0.34 cm after 7 h (p < 0.05). No further change was observed until 12 h (p > 0.05). With starch addition, the height increased to 3.2 cm after 8 h (p < 0.05), then slightly increased to 3.3 cm after 12 h (p > 0.05). With fructose addition, the height increased to 6.7 cm after 11 h (p < 0.05), but slightly decreased after 12 h (p > 0.05). With maltose addition, the height increased to 2.5 cm after 7 h (p < 0.05); however, no change was observed after 8 h (p > 0.05). A slight increase to 2.9 cm was observed at 9 h (p > 0.05). No further change was observed until 12 h. Figure 3. Effect of various sugars on Metschnikowia pulcherrima fermentation. ● Indicates glucose, ■ Indicates sucrose, ◆ Indicates starch, ▲ Indicates fructose, and 〇 Indicates maltose. Note: The fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. Measurements were obtained in triplicate. The error bars indicate the standard deviation. Figure 4 shows the effects of various sugars on C. apicola fermentation. The addition of glucose increased the dough height to 4.9 cm after 10 h (p < 0.05); however, no significant change was observed after 11 h (p > 0.05), followed by a slight increase to 5.2 cm after 12 h (p > 0.05). Sucrose increased the height slightly to 1.6 cm after 2 h (p > 0.05); the height remained unchanged until 3 h (p > 0.05), but increased to 7.7 cm after 12 h (p < 0.05). With starch addition, the height increased to 0.25 cm after 2 h (p < 0.05), remained unchanged until 6 h (p > 0.05), and then increased to 0.81 cm after 8 h (p < 0.05). Thereafter, it remained unchanged until 11 h (p > 0.05) and increased to 1.3 cm after 12 h (p < 0.05). With fructose addition, no change was observed until 4 h (p > 0.05), and the height increased to 1.7 cm after 9 h (p < 0.05). Thereafter, no change was observed until 11 h (p > 0.05). Subsequently, the height increased to 2.2 cm after 12 h (p < 0.05). With maltose addition, the height increased to 0.50 cm after 2 h (p < 0.05), but did not change until 6 h (p > 0.05). Thereafter, it increased slightly to 1.2 cm after 9 h (p > 0.05), remained unchanged until 10 h (p > 0.05), and increased to 2.0 cm after 12 h (p < 0.05). Figure 4. Effect of various sugars on Candida apicola fermentation. ● Indicates glucose, ■ Indicates sucrose, ◆ Indicates starch, ▲ Indicates fructose, and 〇 Indicates maltose. Note: The fermentation capacity is calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. Measurements were obtained in triplicate. The error bars indicate the standard deviation. Agriculture and Food Sciences Research, 2025, 12(2): 116-122 120 © 2025 by the author; licensee Asian Online Journal Publishing Group 3.3. Examination of the Effect of Sugar Concentration Figure 5 shows the effect of glucose concentration on M. pulcherrima fermentation. When 1.0% glucose was added, no change was observed in dough height after 1 hour, but the dough height increased to 6.9 cm after 11 hours (p < 0.05) and remained unchanged until 12 hours (p > 0.05). At a concentration of 2.0%, no change was observed in dough height after 1 hour; however, the height increased to 7.3 cm after 12 hours (p < 0.05). At a concentration of 3.0%, no change was observed after 1 hour (p > 0.05), but after 12 hours, the height increased to 8.7 cm (p < 0.05). At a concentration of 4.0%, the height slightly increased to 0.12 cm after 3 hours (p > 0.05), followed by an increase to 8.7 cm after 12 hours (p < 0.05). Following the addition of 5.0% glucose, the height increased to 10 cm after 12 hours (p < 0.05). Figure 5. Effect of glucose concentration on Metschnikowia pulcherrima fermentation. ● Indicates 1.0%, ■ Indicates 2.0%, ◆ Indicates 3.0%, ▲ Indicates 4.0%, and 〇 Indicates 5.0%. Note: The fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. Measurements were obtained in triplicate. The error bars indicate the standard deviation. Figure 6 shows the effect of sugar concentration on C. apicola fermentation. When sucrose was added at a concentration of 1.0%, no change in dough height was observed after 12 hours. Furthermore, at 2.0%, no change was observed in dough height up to 2 hours, but it increased slightly to 0.57 cm after 3 hours (p > 0.05) and remained unchanged after 12 hours (p < 0.05). At 3.0%, no change was observed after 1 hour, but the height increased slightly to 0.71 cm after 3 hours (p > 0.05) and remained unchanged until 12 hours (p > 0.05). Similarly, no change was observed in dough height at 4.0% until 1 hour; however, after an increase in dough height to 0.71 cm after 2 hours (p < 0.05), no further change was observed until 4 hours (p > 0.05). After 10 hours, the height increased to 6.0 cm (p < 0.05), and no change was observed after 12 hours (p > 0.05). At 5.0%, the height increased to 3.3 cm after 3 hours (p < 0.05), but no change was observed after 4 hours (p > 0.05). Thereafter, it increased to 11 cm after 12 hours (p < 0.05). Figure 6. Effect of sucrose concentration on Candida apicola fermentation. ● Indicates 1.0%, ■ Indicates 2.0%, ◆ Indicates 3.0%, ▲ Indicates 4.0%, and 〇 Indicates 5.0%. Note: The fermentation capacity was calculated as the increase in dough height per 0.10 g of yeast per hour; the results are shown as integrated value. Measurements were obtained in triplicate. The error bars indicate the standard deviation. 4. Discussion 4.1. Examination of the Optimal Fermentation Temperature for Each Yeast In this study, the optimal temperature, sugar type, and concentration required for fermentation using M. pulcherrima and C. apicola as natural yeasts in bread production were investigated. After 5 hours of fermentation, M. pulcherrima and C. apicola showed high fermentation capacities at 25°C and 35°C, respectively; however, after 5 hours, they exhibited relatively high fermentation capacities at 30°C and 35°C, respectively. Metschnikowia pulcherrima Agriculture and Food Sciences Research, 2025, 12(2): 116-122 121 © 2025 by the author; licensee Asian Online Journal Publishing Group grows efficiently at temperatures in the range of 15–20°C [2]; whereas, C. apicola grows efficiently at 25–37 °C [12], indicating that it does not grow well in low-temperature environments. Metschnikowia pulcherrima has been isolated as a dominant fermentation species during the fermentation of Fiano di Avellino grapes at 9 °C in wine production [24], suggesting that it can survive at low temperatures and show high fermentation potential. In addition, C. apicola can survive at low temperatures while marginally maintaining its fermentation potential. Detailed reports on the optimal fermentation temperature of C. apicola are lacking. As a species similar to C. apicola, C. tropicalis has been reported to be heat-resistant and capable of producing ethanol from xylose [25]. Candida species may have a high fermentation capacity at relatively high temperatures, suggesting that it is difficult for C. apicola to ferment substrates at lower temperatures. 4.2. Investigation of the Optimal Sugar Type for Each Yeast In this study, M. pulcherrima and C. apicola showed high fermentation capacities following the addition of glucose and sucrose, respectively. Glucose, a monosaccharide, is used by yeast in ethanol fermentation. In a previous study, 31 types of S. cerevisiae isolates from palm wine were provided with glucose, galactose, fructose, sucrose, maltose, trehalose, and raffinose, either in combination or individually, to ferment a substrate, and glucose was used for fermentation in all tests [26]. Among the aforementioned sugars, glucose, galactose, and fructose are monosaccharides; sucrose, maltose, and trehalose are disaccharides; and raffinose is the only trisaccharide. As these disaccharides and the trisaccharide contain glucose, they can be fermented if the yeast produces enzymes that can break them down into glucose. Reportedly, the strength of sucrase activity varies among different species of S. cerevisiae [27]. Hence, the type of sugar available and the fermentation rate may differ among different yeast species. In this study, the fermentation capacity of M. pulcherrima for sucrose was weaker than that for maltose and starch (Figure 3). Metschnikowia pulcherrima uses glucose and not sucrose in the fermentation of soy whey and produces ethanol as a product [28-30], consistent with the findings of the present study, suggesting that M. pulcherrima does not produce sucrase or possesses weak sucrase activity. Alcohol fermentation has been reported to be slow with maltose [31], and as starch is maltose-bound, M. pulcherrima may have weak maltase and amylase activities. Candida apicola has weak maltase or amylase activity (Figure 4) and shows high fermentation efficiency for sucrose. In fermentation capacity tests by sugar type, C. apicola was reportedly able to ferment sucrose but not fructose [12]. Similar results were obtained in this study. Hence, C. apicola has high sucrase activity; however, it was also able to ferment the dough with fructose in this study. The glucose and fructose produced from sucrose through sucrase were used for fermentation. However, the slow onset of fermentation of glucose and fructose suggests that sucrose may promote fermentation by C. apicola. In addition, as C. apicola has been reported to tolerate high ethanol concentrations [21], it is possible that ethanol produced through fermentation promoted the fermentation of glucose and fructose. However, baking bread was the focus of this study, and therefore, the fermentation mechanism of these yeasts in baking is a subject for future research. 4.3. Examination of the Effect of Sugar Concentration Metschnikowia pulcherrima and C. apicola showed the maximum fermentation efficiency at a sugar concentration of 5.0%. During fermentation, the fermentation capacity of C. apicola was affected by sugar concentration. In a previous study, fermentation by S. cerevisiae increased ethanol concentration with increasing glucose concentrations from 2.0% to 8.0% [32]. Furthermore, as Starmerella zemplinina (synonym Candida zemplinina) has been reported to grow faster when glucose is added at concentrations of 2.0% and 20% [21], fermentation efficiency may have increased with increasing glucose concentrations up to 20%. In contrast, the fermentation efficiency of Hanseniaspora guilliermondii reportedly decreases at sugar concentrations >300 g/L [14]. These results indicate that the effect of sugar concentration on fermentation varies depending on the yeast type. The yeasts investigated in this study showed the highest fermentation efficiency at a 5.0% sugar concentration, and it is highly probable that the efficiency can be increased by increasing the sugar concentration. However, as a considerable increase in sugar concentration significantly affects the flavor of bread, the effect of sugar concentration on flavor must be considered. This study has some limitations. As both wheat and water are used in bread production, the effects of wheat and the ions in water should be considered. However, these factors were not examined in this study; therefore, their influence on the fermentation capacity of M. pulcherrima and C. apicola should be investigated in future studies to clarify the conditions suitable for bread production. Moreover, the fermentation capacity of natural yeasts could change depending on the environment. The present study did not focus on this aspect, warranting future studies on the effects of environmental factors on the fermentation capacity of these yeasts. 5. Conclusion The effects of temperature, sugar type, and concentration on yeast fermentation capacity were investigated in this study. The optimal fermentation temperature was 25°C for M. pulcherrima and 35°C for C. apicola. The concentration and type of sugar that most enhanced the fermentation efficiency of each yeast were 5.0% glucose for M. pulcherrima and 5.0% sucrose for C. apicola. Overall, fermentation using M. pulcherrima and C. apicola under appropriate temperature and sugar concentration conditions could potentially enhance the efficiency of bread production. Given the limited research examining the use of these natural yeasts in food, further studies are required to ensure their safety, effectiveness, and hygienic handling. References [1] F. Kayadelen, B. Agirman, N. P. Jolly, and H. Erten, "The influence of Torulaspora delbrueckii on beer fermentation," FEMS Yeast Research, vol. 23, pp. 1-13. 2023. https://doi.org/10.1093/femsyr/foad006 [2] A. Morata, I. Loira, C. Escott, J. M. del Fresno, M. A. Bañuelos, and J. A. Suárez-Lepe, "Applications of Metschnikowia pulcherrima in wine biotechnology," Fermentation, vol. 5, no. 3, p. 63, 2019. https://doi.org/10.3390/fermentation5030063 [3] G. M. Walker, P. Lappe-Oliveras, R. Moreno-Terrazas C, M. Kirchmayr, M. Arellano-Plaza, and A. C. Gschaedler-Mathis, Yeasts associated with the production of distilled alcoholic beverages, Yeasts in the Production of Wine. Berlin, Germany: Springer Nature, 2019, pp. 477-512. https://doi.org/10.1093/femsyr/foad006 https://doi.org/10.3390/fermentation5030063 Agriculture and Food Sciences Research, 2025, 12(2): 116-122 122 © 2025 by the author; licensee Asian Online Journal Publishing Group [4] M. Yamada, K. Koizumi, N. Akaishi, and M. Mineki, "Effect of different yeasts on bread quality," Katei Gahou, vol. 72, no. 12, pp. 796- 807, 2021. [5] M. Heitmann, E. Zannini, C. Axel, and E. Arendt, "Correlation of flavor profile to sensory analysis of bread produced with different Saccharomyces cerevisiae originating from the baking and beverage industry," Cereal Chemistry, vol. 94, no. 4, pp. 746-751. 2017. https://doi.org/10.1094/CCHEM-03-17-0044-R [6] S. Kato, "A case of wild yeast starter from raisins, in which red koji mold Monascus ruber was identified along with Saccharomyces cerevisiae," New Food Industry, vol. 66, no. 2, pp. 77–85, 2024. [7] Y. Oda, H. Yamauchi, and M. Tamura, "Development of baker's yeast" Tokachino" through collaboration of industry, academia and government," Journal of the Japanese Society for Food Science and Technology, vol. 59, no. 1, pp. 1–5, 2012. [8] T. Hisatomi and K. Toyomura, "Isolation, identification, and characterization of wild budding yeasts from rose flowers in Fukuyama city, Hiroshima, Japan, and their application in bread and wine production," Mycoscience, vol. 62, no. 6, pp. 382-389. 2021. https://doi.org/10.47371/mycosci.2021.10.003 [9] A. N. M. Mamun-Or-Rashid, T. T. Lucy, and M. K. Pramanik, "Isolation, identification, optimization of baker’s yeast from natural sources, scale-up production using molasses as a cheap carbohydrate source, and evaluation for bread production," Applied Microbiology, vol. 2, no. 3, pp. 516-533, 2022. https://doi.org/10.3390/applmicrobiol2030040 [10] Y. Aoki, "Research report on natural yeast," Research Bulletin, vol. 42, pp. 45–50, 2017. https://doi.org/10.20582/nfcc.42.0_45 [11] M. Kamakura and M. Mayama, "Bread making property of Hanseniaspora meyeri, one of the wild yeast isolates from petal of Citrus sudachi," Bulletin of Shikoku University, vol. 34, pp. 37-46, 2012. [12] Z. Tsegaye, G. Tefera, B. Gizaw, and E. Abatenh, "Characterization of yeast species isolated from local fruits used for bakery industrial application," Journal of Applied Microbiological Research, vol. 1, no. 1, pp. 21-26, 2018. [13] A. Ohashi, K. Fukuyama, and S. Ohba, "A measurement of the optimum temperature for alcoholic fermentation," Hiyoshi Review of Natural Science, vol. 45, pp. 1-13, 2009. [14] V. M. Vučurović, V. S. Puškaš, and U. D. Miljić, "Bioethanol production from sugar beet molasses and thick juice by free and immobilised Saccharomyces cerevisiae," Journal of the Institute of Brewing, vol. 125, no. 1, pp. 134-142. 2019. https://doi.org/10.1002/jib.536 [15] M. Nakatani and H. Seki, "Cancer cell growth inhibitory, anti-oxidant, and α-glucosidase inhibitory effects of enzyme syrup obtained from apple fermentation," Functional Food Research, vol. 16, pp. 65–74, 2020. [16] R. Hashimoto and H. Seki, "The study of functionality in syrup obtained from kiwi fruit fermentation," Functional Food Research, vol. 17, pp. 118–125, 2021. [17] L. M. Gerard, M. B. Corrado, C. V. Davies, C. A. Soldá, M. G. Dalzotto, and S. Esteche, "Isolation and identification of native yeasts from the spontaneous fermentation of grape musts," Archives of Microbiology, vol. 205, no. 9, p. 302. 2023. https://doi.org/10.1007/s00203-023-03646-1 [18] R. Li, D. Feng, H. Wang, Z. Zhang, N. Li, and Y. Sun, "Genetic diversity of non-Saccharomyces yeasts associated with spontaneous fermentation of Cabernet Sauvignon wines from Ningxia, China," Frontiers in Microbiology, vol. 14, p. 1253969. 2023. https://doi.org/10.3389/fmicb.2023.1253969 [19] A. Boban, V. Milanović, M. V. Bratinčević, C. Botta, I. Ferrocino, F. Cardinali, S. Ivić, G. Rampanti, and I. Budić-Leto, "Spontaneous fermentation of Maraština wines: The correlation between autochthonous mycobiota and phenolic compounds," Food Research International, vol. 180, p. 114072. 2024. https://doi.org/10.1016/j.foodres.2024.114072 [20] H. Abriouel, N. Benomar, R. Lucas, and A. Gálvez, "Culture-independent study of the diversity of microbial populations in brines during fermentation of naturally-fermented Aloreña green table olives," International Journal of food Microbiology, vol. 144, no. 3, pp. 487-496. 2011. https://doi.org/10.1016/j.ijfoodmicro.2010.11.006 [21] R. Tofalo, C. Chaves-López, F. Di Fabio, M. Schirone, G. E. Felis, S. Torriani, A. Paparella, and G. Suzzi, "Molecular identification and osmotolerant profile of wine yeasts that ferment a high sugar grape must," International Journal of Food Microbiology, vol. 130, no. 3, pp. 179-187. 2009. https://doi.org/10.1016/j.ijfoodmicro.2009.01.024 [22] E. S. Oliveira, C. A. Rosa, M. A. Morgano, and G. E. Serra, "Fermentation characteristics as criteria for selection of cachaça yeast," World Journal of Microbiology and Biotechnology, vol. 20, no. 1, pp. 19-24. 2004. https://doi.org/10.1023/B:WIBI.0000013286.30695.4e [23] E. S. Oliveira, H. M. A. B. Cardello, E. M. Jeronimo, E. L. R. Souza, and G. E. Serra, "The influence of different yeasts on the fermentation, composition and sensory quality of cachaça," World Journal of Microbiology and Biotechnology, vol. 21, no. 5, pp. 707-715. 2005. https://doi.org/10.1007/s11274-004-4490-4 [24] E. Petruzziello, G. Blaiotta, E. Pittari, P. Piombino, and M. Aponte, "Isolation and characterization of cryotolerant yeasts from Fiano di Avellino grapes fermented at low temperatures," Foods, vol. 12, no. 3, p. 526. 2023. https://doi.org/10.3390/foods12030526 [25] J. E. Nweze, I. Ndubuisi, Y. Murata, H. Omae, and J. C. Ogbonna, "Isolation and evaluation of xylose-fermenting thermotolerant yeasts for bioethanol production," Biofuels, vol. 12, no. 8, pp. 961–970, 2021. https://doi.org/10.1080/17597269.2018.1564480 [26] C. Y. Tra Bi, C. A. Kouakou-Kouamé, F. K. N’guessan, M. K. Djè, and D. Montet, "Phenotypic characterization of indigenous Saccharomyces cerevisiae strains associated with sorghum beer and palm wines," World Journal of Microbiology and Biotechnology, vol. 37, no. 2, p. 24. 2021. https://doi.org/10.1007/s11274-020-02990-4 [27] C.-Y. Zhang, X. Lin, B. Feng, X.-E. Liu, X.-W. Bai, J. Xu, L. Pi, and D.-G. Xiao, "Enhanced leavening properties of baker’s yeast by reducing sucrase activity in sweet dough," Applied Microbiology and Biotechnology, vol. 100, no. 14, pp. 6375-6383. 2016. https://doi.org/10.1007/s00253-016-7449-0 [28] J.-Y. Chua, Y. Lu, and S.-Q. Liu, "Evaluation of five commercial non-Saccharomyces yeasts in fermentation of soy (tofu) whey into an alcoholic beverage," Food Microbiology, vol. 76, pp. 533-542. 2018. https://doi.org/10.1016/j.fm.2018.07.016 [29] R. Rodríguez Madrera, R. Pando Bedriñana, and B. Suárez Valles, "Evaluation of indigenous non-Saccharomyces cider yeasts for use in brewing," European Food Research and Technology, vol. 247, no. 4, pp. 819-828. 2021. https://doi.org/10.1007/s00217-020-03665- y [30] Y. Lu, D. Huang, P. R. Lee, and S. Q. Liu, "Assessment of volatile and non‐volatile compounds in durian wines fermented with four commercial non‐Saccharomyces yeasts," Journal of the Science of Food and Agriculture, vol. 96, no. 5, pp. 1511-1521. 2016. https://doi.org/10.1002/jsfa.7253 [31] D. Einfalt, "Barley-sorghum craft beer production with Saccharomyces cerevisiae, Torulaspora delbrueckii and Metschnikowia pulcherrima yeast strains," European Food Research and Technology, vol. 247, no. 2, pp. 385-393, 2021. https://doi.org/10.1007/s00217-020-03632-7 [32] S.-Y. Tsai, Y.-C. Hsu, C.-M. Shu, K.-H. Lin, and C.-P. Lin, "Synchronization of isothermal calorimetry and liquid cultivation identifying the beneficial conditions for producing ethanol by yeast Saccharomyces cerevisiae fermentation," Journal of Thermal Analysis and Calorimetry, vol. 142, no. 2, pp. 829-840. 2020. https://doi.org/10.1007/s10973-020-09629-4 Asian Online Journal Publishing Group is not responsible or answerable for any loss, damage or liability, etc. caused in relation to/arising out of the use of the content. Any queries should be directed to the corresponding author of the article. https://doi.org/10.1094/CCHEM-03-17-0044-R https://doi.org/10.47371/mycosci.2021.10.003 https://doi.org/10.3390/applmicrobiol2030040 https://doi.org/10.20582/nfcc.42.0_45 https://doi.org/10.1002/jib.536 https://doi.org/10.1007/s00203-023-03646-1 https://doi.org/10.3389/fmicb.2023.1253969 https://doi.org/10.1016/j.foodres.2024.114072 https://doi.org/10.1016/j.ijfoodmicro.2010.11.006 https://doi.org/10.1016/j.ijfoodmicro.2009.01.024 https://doi.org/10.1023/B:WIBI.0000013286.30695.4e https://doi.org/10.1007/s11274-004-4490-4 https://doi.org/10.3390/foods12030526 https://doi.org/10.1080/17597269.2018.1564480 https://doi.org/10.1007/s11274-020-02990-4 https://doi.org/10.1007/s00253-016-7449-0 https://doi.org/10.1016/j.fm.2018.07.016 https://doi.org/10.1007/s00217-020-03665-y https://doi.org/10.1007/s00217-020-03665-y https://doi.org/10.1002/jsfa.7253 https://doi.org/10.1007/s00217-020-03632-7 https://doi.org/10.1007/s10973-020-09629-4