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For the first time, the dual effect of coffee cherry sanitization methods to control the microbial load in processing and the influence of fermentation time on coffee quality parameters was evaluated. Two assays were carried out by wet processing: I) Sanitization of the coffee cherry (ST1: Unclassified processed cherries; ST2: Classified and sanitized cherries with drinking water; ST3: Classified and sanitized cherries with a chemical agents and II) Fermentation times (FT1: 12 h; FT2: 24 h; FT3: 48 h; FT4: 72 h and FT5: 96 h). pH, temperature, and dissolved oxygen were monitored during fermentation. Counts of Lactic Acid Bacteria - LAB, mesophiles, and yeasts were carried out on the coffee mass before and after fermentation. Caffeine and chlorogenic acid contents were determined by HPLC-DAD and the sensory profile by methodology for specialty coffees (SCA). The main findings showed that: sanitization with Timsen® did not significantly influence the evolution of pH during fermentation (p > 0.05), but it can reduce to a small extent the action of LAB at the end of the process. It was observed that the temperature of the coffee mass tends to balance with the ambient temperature, with significant effects (p < 0.05) of sanitization (ST2 and ST3) on the stability of this variable during fermentation. Timsen® as a disinfectant affected microbial populations and improved the sensory profile in the cup. In prolonged coffee fermentations (FT3, FT4 and FT5), the pH of the coffee mass tended to stabilize after 36 h, regardless of the process time. Likewise, a correlation was evident between a higher microbial load correlated with better sensory profiles in FT4 and FT5. Neither the sanitization process nor the fermentation time significantly affected the caffeine and chlorogenic acid contents of the coffee, both in its green and roasted states. Consequently, the sanitization of cherry coffee with Timsen® and prolonged fermentation times favor the safety and coffee final quality in the cup.

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Research into microbial interactions during coffee processing is essential for developing new methods that adapt to climate change and improve flavor, thus enhancing the resilience and quality of global coffee production. This study aimed to investigate how microbial communities interact and contribute to flavor development in coffee processing within humid subtropical climates. Employing Illumina sequencing for microbial dynamics analysis, and high-performance liquid chromatography (HPLC) integrated with gas chromatography-mass spectrometry (GC-MS) for metabolite assessment, the study revealed intricate microbial diversity and associated metabolic activities. Throughout the fermentation process, dominant microbial species included Enterobacter, Erwinia, Kluyvera, and Pantoea from the prokaryotic group, and Fusarium, Cladosporium, Kurtzmaniella, Leptosphaerulina, Neonectria, and Penicillium from the eukaryotic group. The key metabolites identified were ethanol, and lactic, acetic, and citric acids. Notably, the bacterial community plays a crucial role in flavor development by utilizing metabolic versatility to produce esters and alcohols, while plant-derived metabolites such as caffeine and linalool remain stable throughout the fermentation process. The undirected network analysis revealed 321 interactions among microbial species and key substances during the fermentation process, with Enterobacter, Kluyvera, and Serratia showing strong connections with sugar and various volatile compounds, such as hexanal, benzaldehyde, 3-methylbenzaldehyde, 2-butenal, and 4-heptenal. These interactions, including inhibitory effects by Fusarium and Cladosporium, suggest microbial adaptability to subtropical conditions, potentially influencing fermentation and coffee quality. The sensory analysis showed that the final beverage obtained a score of 80.83 ± 0.39, being classified as a specialty coffee by the Specialty Coffee Association (SCA) metrics. Nonetheless, further enhancements in acidity, body, and aftertaste could lead to a more balanced flavor profile. The findings of this research hold substantial implications for the coffee industry in humid subtropical regions, offering potential strategies to enhance flavor quality and consistency through controlled fermentation practices. Furthermore, this study contributes to the broader understanding of how microbial ecology interplays with environmental factors to influence food and beverage fermentation, a topic of growing interest in the context of climate change and sustainable agriculture.

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Background: Mild Colombian coffees are recognized worldwide for their high-quality coffee cup. However, there have been some failures in post-harvest practices, such as coffee grain fermentation. These failures could occasionally lead to defects and inconsistencies in quality products and economic losses for coffee farmers. In Colombia, one of the fermentation methods most used by coffee growers is wet fermentation, conducted by submerging the de-pulped coffee beans for enough time in water tanks to remove the mucilage. Objectives: We evaluated the effect of the water (g)/de-pulped coffee (g) ratio (I: 0/25, II: 10/25, III: 20/25) and final fermentation time (24, 48, and 72 hours) on the total number of microbial groups. We also identified microorganisms of interest as starter cultures. Methods: We used a completely randomized experimental design with two factors; the effect of the water (g)/de-pulped coffee (g) ratio (I: 0/25, II: 10/25, III: 20/25) and final fermentation time (24, 48, and 72 hours), for 9 treatments with two replicates. During the coffee fermentation (1,950 g), the pH and °Brix were monitored. Total counts of different microbial groups (mesophiles, coliforms, lactic-acid bacteria, acetic-acid bacteria, and yeasts) were performed. Various isolates of microorganisms of interest as starter cultures (lactic-acid bacteria and yeasts) were identified using molecular sequencing techniques. Results: 21 lactic-acid bacteria (LAB) isolates and 22 yeasts were obtained from the different mini-batch fermentation systems. The most abundant lactic-acid bacteria species found were Lactiplantibacillus plantarum (46%) and Levilactobacillus brevis (31%). Pichia kluivery (39%) and Torulaspora delbrueckii (22%) were the most abundant yeast species. Conclusion The studied factors did not have effect over the microorganism's development. The identified bacterial and yeasts species have potential as starter cultures for better-quality coffees and in fermentation-related applications.

Antecedentes: Los cafés suaves lavados colombianos son reconocidos a nivel mundial por su buena puntuación sensorial; sin embargo, se han detectado fallas en las prácticas de postcosecha, como lo es la fermentación de los granos de café. Dichas fallas pueden causar defectos y carecer de consistencia en la calidad del producto, ocasionando pérdidas económicas para los caficultores. En Colombia, uno de los métodos más usados por los caficultores es la fermentación húmeda, la cual consiste en sumergir los granos de café despulpado en tanques con agua por un período de tiempo que permita la remoción del mucílago. Objetivos: La presente investigación evaluó la incidencia que tienen la proporción agua/granos despulpados de café (I: 0/25, II: 10/25, III: 20/25) y el tiempo final de fermentación (24, 48 y 72 horas) en el recuento final de grupos microbianos. Por otra parte, se identificaron taxonómicamente microorganismos de interés para su uso como cultivos iniciadores. Métodos: Mini-lotes consistieron en café despulpado (1950 g) puesto en recipientes de plástico abiertos y sumergidos en agua. Se aplicó un diseño experimental completamente aleatorizado de dos factores (proporción agua/ granos de café despulpado y tiempo) a tres niveles, para un total de nueve tratamientos con dos replicas. Durante las fermentaciones de café (1,950 g), el pH y los grados ºBrix, fueron monitoreados. Se realizaron los recuentos totales de los diferentes grupos microbianos: mesófilos, coliformes, bacterias ácido-lácticas, bacterias ácido-acéticas y levaduras. Se identificaron molecularmente diferentes aislados con potencial para ser usados como cultivos iniciadores (bacterias ácido-lácticas y levaduras). Resultados: Los resultados obtenidos mostraron que no hubo diferencia estádisticamente significativa entre los tratamientos aplicados y el recuento final de microorganismos. Un total de 21 aislados de bacterias ácido-lácticas (BAL) y 22 levaduras lograron obtenerse a partir de los diferentes sistemas de fermentación en mini-lote. Las especies de bacterias ácido-lácticas con mayor porcentaje acorde a su identificación taxonómica, corresponden a Lactiplantibacillus plantarum (46%), Levilactobacillus brevis (31%). Las especies de levaduras con mayor porcentaje acorde a su identificación taxonómica corresponden a Pichia kluivery (39%) y Torulaspora delbrueckii (22%). Conclusión Los factores estudiados no afectaron el crecimiento de ninguno de los grupos microbianos presentes en la fermentacion del café. Las especies de microorganismos identificados tienen potencial para se usados como cultivos starter o en aplicaciones dentro de las ciencias de fermentación.

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Variation in fermentation time may be an essential alternative to provide coffee beverages with different and unique sensory profiles. This work investigated the microbiological, chemical, and sensory changes in coffees submitted to different fermentation durations (0, 24, 48, 72, and 96 h). Self-induced anaerobiosis fermentation (SIAF) was used, and two treatments were performed: spontaneous fermentation and inoculation with S. cerevisiae CCMA0543. Microbiological analyses were performed, and the permanence of the inoculum was monitored. Chromatography (sugars, organic acids, and volatile compounds) was analyzed, and sensory analysis (temporal dominance of sensations - TDS) was performed. A total of 228 isolates were identified during spontaneous fermentation. The dominant bacteria and yeasts were Leuconostoc mesenteroides, Lactiplantibacillus plantarum, Staphylococcus warneri, Bacillus sp., Torulaspora delbrueckii, Hanseniaspora uvarum, and Meyerozyma caribbica. High concentrations of citric (18.67 mg.g- 1) and succinic (5.04 mg.g- 1) acids were detected at 96 h in SIAF fermentation. One hundred twenty-one volatile compounds were detected, but 22 were detected only in inoculated coffees. In spontaneous fermentation, 48 h of fermentation showed woody notes, while 72 h showed chestnuts. However, in the inoculated coffee, 72 h of fermentation showed high fruity dominance, and 96 h of fermentation was the only one with herbaceous notes. In addition, yeast inoculation increased the intensity of caramel notes in the first 48 h and increased the fruity flavor after 72 h of fermentation. Therefore, the type of fermentation (with or without inoculation) and the chosen fermentation time will depend on the sensorial profile the producer intends to obtain.

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Fermentation, a critical post-harvest process, can be strategically manipulated to augment coffee quality. This enhancement is achieved through the activity of microorganisms, which generate metabolites instrumental in the formation of distinct sensory profiles. This study investigated the impact of different fermentation methods on the quality of coffee beverages, specifically utilizing the Catiguá MG2 variety. The experimental setup involved fermenting the coffee in 200 L bioreactors, employing both natural and pulped coffee beans. The fermentation process utilized was self-induced anaerobic fermentation (SIAF), conducted in either a solid-state or submerged medium over a 96 h period. Analytical sampling was conducted initially and at 24 h intervals thereafter to quantify the concentration of sugars, alcohols, and organic acids. Sensory evaluation was performed using the established protocols of the Specialty Coffee Association (SCA). The outcomes of this investigation reveal that fermentation substantially enhances the quality of coffee, with each treatment protocol yielding divergent profiles of acids and alcohols, thereby influencing the sensory characteristics of the resulting beverage. Notably, superior quality beverages were produced from naturally processed coffee subjected to solid-state fermentation for durations exceeding 24 h. These findings underscore the significant influence of fermentation techniques and duration on the sensory attributes and overall quality of coffee.

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This study has innovative aspects related to the use of sequential inoculation technique in the coffee bean fermentation process: the inoculation of Lactiplantibacillus plantarum followed by Saccharomyces cerevisiae, in the fermentation of coffee fruit for the production of specialty natural coffees. The objective was to evaluate the effect of this technique and of the total fermentation time on the sensory attributes of the coffee beverage and on the organic acid profile, bioactive compounds, and fatty acid profile of the beans. The fermentation of coffee fruit with sequential inoculation resulted in greater acidity of the beverage and contributed to increases of up to 2 points in coffee fermented. The total fermentation time was directly related to the organic acid content, and the longer the total fermentation time was, the greater the organic acid content. The fatty acid content and bioactive compound content showed little variation among treatments.

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This study explores the variances in the organic, chemical, and sensory attributes of fermented coffee beans, specifically examining how post-harvest processes influence cup quality. Coffee fruits from the Catuaí IAC-144 variety were processed using both natural coffee (NC) and pulped coffee (PC) methods. The fruits were then subjected to self-induced anaerobic fermentation (SIAF) using one of the following fermentation methods: solid-state fermentation (SSF) or submerged fermentation (SMF). Within these methods, either spontaneous fermentation (SPF) or starter culture fermentation (SCF) was applied. Each method was conducted over periods of 24, 48, and 72 h. For this purpose, two-hundred-liter bioreactors were used, along with two control treatments. Numerous parameters were monitored throughout the fermentation process. A comprehensive chemical profiling and sensory analysis, adhering to the guidelines of the Specialty Coffee Association, were conducted to evaluate the influence of these fermentation processes on the flavor, aroma, and body characteristics of the coffee beverage across multiple dimensions. Data analysis and predictive modeling were performed using machine learning techniques. This study found that NC exhibited a higher production of acids (citric, malic, succinic, and lactic) compared to PC, resulting in distinct chemical and sensory profiles. The decision tree showed that fructose and malic and succinic acids were identified as the main factors enhancing sensory notes during cupping. SMF promoted higher concentrations of lactic acid, while SSF led to increased ethanol content. Consequently, the SIAF process enhances the sensory quality of coffee, adding value to the product by generating diverse sensory profiles.

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Using starter culture in liquid form is not economically viable in the coffee fermentation process. This work aimed to compare the fermentative performances of fresh and microencapsulated yeasts in coffee under self-induced anaerobic fermentation (SIAF). The inoculum permanence was monitored, and sugars, alcohols, acids, and volatile compounds were analyzed by chromatography. In addition, sensory analysis was performed on roasted beans. After 180 h of fermentation in the natural process, microencapsulated Torulaspora delbrueckii (MT) (7.97 × 107 cells/g) showed a higher population thanfresh Torulaspora delbrueckii (FT) (1.76 × 107 cells/g). The same acids and volatile compounds were detected in coffees with fresh and microencapsulated yeast. However, the yeast state influenced the concentration of the compounds. In pulped coffee, the coffee inoculated withmicroencapsulated Saccharomyces cerevisiae (MS) obtained the highest concentration of alcohols, esters, pyrazines, and others compared with fresh Saccharomyces cerevisiae (FS), with an increase of up to 47%. Furthermore, the coffee inoculated with MT obtained the highest concentration in almost all chemical classes in both processes compared with FT. These differences ranged up to 55%. Regarding sensory analysis, coffees inoculated with MS showed dominant notes of fruity, caramel, and nuts in the natural process. Otherwise, in pulped process, coffees inoculated with MT showed caramel, honey, and nuts. Therefore, the microencapsulated yeasts were metabolically active and may be considered with commercial potential. Considering the parameters analyzed, the most suitable yeast for natural and pulped processing would be MS and MT, respectively.

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Altitude changes the coffee fruits and beans composition before and after harvesting. We aimed to evaluate the effect of altitude on the microbial community structure associated with pulped coffee fruits under self-induced anaerobic fermentation (SIAF) and their acids, volatiles, and antioxidants biochemical profiles. The most abundant bacterial genera were Gluconobacter (800 m), Weissella (1,000 m), and Leclercia (1,200 and 1,400 m). Yeasts dominated the pulped natural fermentations within the fungal species, containing high abundances of Cystofilobasidium infirmominiatum, Wickerhamomyces anomalus, and Meyerozyma caribbica. Citric, alcohols, and caffeine were the most dominant compounds in SIAF among acids, volatiles, chemical groups, and antioxidants. High altitude coffees favor alcohols, aldehydes, and esters groups, while low altitude coffees favor phenols.

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The storage of microorganisms in liquid form is the main drawback of commercializing epiphytic coffee yeasts. This work aimed to evaluate the fermentative performance of microencapsulated yeasts by spray drying in a coffee peel and pulp media (CPM). The yeasts, Saccharomyces cerevisiae CCMA 0543, Torulaspora delbrueckii CCMA 0684, and Meyerozyma caribbica CCMA 1738, were microencapsulated using maltodextrin DE10 (MD), high maltose (MA), and whey powder (WP) as wall materials. A Central Composite Rotational Design (CCRD) was used to investigate the effect of operating parameters on the microcapsules' cell viability, drying yield, and water activity. Yeasts reached cell viability and drying yields above 90 and 50 %, respectively. WP maintained the cell viability of the three yeasts over 90 days of storage at room temperature (25 °C) and was selected as a wall material for the three yeasts. M. caribbica showed to be more sensitive to spray drying and less resistant to storage. Some differences were found in the fermentation of the CPM medium, but the microencapsulated yeasts maintained their biotechnological characteristics. Therefore, the microencapsulation of epiphytic coffee yeasts by spray drying was promising to be used in the coffee fermentation process.

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In this study, an investigation of the microbial community structure and chemical changes in different layers of a static coffee beans fermentation tank (named self-induced anaerobic fermentation-SIAF) was conducted at different times (24, 48, and 72 h). The microbial taxonomic composition comprised a high prevalence of Enterobacteriaceae and Nectriaceae and low prevalence of lactic acid bacteria and yeast, which greatly differs from the traditional process performed in open tanks. No major variation in bacterial and fungal diversity was observed between the bottom, middle, and top layers of the fermentation tank. On the other hand, the metabolism of these microorganisms varied significantly, showing a higher consumption of pulp sugar and production of metabolites in the bottom and middle layers compared to the top part of the fermentation tank. Extended processes (48 and 72 h) allowed a higher production of key-metabolites during fermentation (e.g., 3-octanol, ethyl acetate, and amyl acetate), accumulation in roasted coffee beans (acetic acid, pyrazine, methyl, 2-propanone, 1-hydroxy), and diversification of sensory profiles of coffee beverages compared to 24 h of fermentation process. In summary, this study demonstrated that SIAF harbored radically different dominant microbial groups compared to traditional coffee processing, and diversification of fermentation time could be an important tool to provide coffee beverages with novel and desirable flavor profiles.

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Coffee harvested in the Caparaó region (Minas Gerais, Brazil) is associated with high-quality coffee beans resulting in high-quality beverages. We characterize, microbiologically and chemically, fermented coffees from different altitudes through target NGS, chromatography, and conventional chemical assays. The genera Gluconobacter and Weissella were dominant in coffee's fruits from altitudes 800 and 1,000 m. Among the Eukaryotic community, yeasts were the most dominant in all altitudes. The most dominant fungal genus was Cystofilobasidium, which inhabits cold environments and resists low temperatures. The content of acetic acid was higher at altitudes 1,200 and 1,400 m. Lactic acid and the genus Leuconostoc (Pearson: 0.93) were positively correlated. The relative concentration of volatile alcohols, especially of 2-heptanol, was high at all altitudes. Bacteria population was higher in coffees from 800 m, while at 1,000 m, fungi richness was favored. The altitude is an important variable that caused shifts in the microbial community and biochemical compounds content, even in coffees belonging to the same variety and cultivated in the same region under SIAF (self-induced anaerobic fermentation) conditions. Coffee from lower altitudes has higher volatile alcohols content, while high altitudes have esters, aldehydes, and total phenolics contents.

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Starter cultures during fermentation of Coffea arabica var. Mundo Novo processed in open stainless-steel vessels by natural and pulped natural methods were studied. The yeasts Meyerozyma caribbica (CCMA 0198), Saccharomyces cerevisiae (CCMA 0543), Candida parapsilosis (CCMA 0544), and Torulaspora delbrueckii (CCMA 0684) were inoculated separately in two different coffee processes: natural and pulped natural. The qPCR (real-time quantitative polymerase chain reaction) was used as a culture-independent method to monitor the inoculum's permanence. Changes in microbial metabolites (organic acids and volatile) production were evaluated by high-performance liquid chromatography (HPLC) and gas chromatograph-mass spectrometry (GC-MS), respectively. The sensory analysis was assessed in roasted beans. The fermentation lasted 27 h, and the coffee temperature ranged from 16.5 to 24.0 °C. The starter culture population was dominant throughout fermentation. S. cerevisiae (CCMA 0543) and T. delbrueckii (CCMA 0684) presented a higher population in natural processing. However, in pulped natural processing, M. caribbica (CCMA 0198) and C. parapsilosis (CCMA 0544) were the dominant populations. Citric, malic, and succinic acids were naturally present in coffee. Lactic, isobutyric, and isovaleric acids were detected at the end of the fermentation in different treatments. Lactic acid was detected in samples at the end of fermentation in Control and CCMA 0198 treatment. NAT coffee inoculated with CCMA 0684 presented isobutyric acid and isovaleric acid concentrations. Volatile compounds, such as 2,6-diethylpyrazine was detected in treatments inoculated with yeasts, but not in Controls. 2-acetoxymethylfuran was only detected in samples inoculated with CCMA 0198 from both NAT and PN methods. Samples fermented with S. cerevisiae (CCMA 0543) presented the highest sensorial scores in both processing (84.75 and 84.92). The inoculated coffee beans showed higher scores of sweetness, long aftertaste, and greater complexity. The starter cultures influenced the sensorial profiles through the synthesis of specific volatile constituents. However, considering all parameters analyzed, S. cerevisiae (CCMA 0543) would be the most suitable yeast for the var. Mundo Novo processed by both fermentation methods.

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The use of starter cultures during food fermentation aims to standardize the process and to obtain a higher quality product. The objectives were to study mesophilic bacteria (MB) and lactic acid bacteria (LAB) isolated from wet coffee processing and evaluate their performance in a pulped coffee medium. Eighty-six bacteria isolates (59 MB and 27 LAB) were assessed for pectinolytic activity, metabolite production, and pH value decrease in coffee-based culture (CPM). Seven bacteria strains (3 MB and 4 LAB) were selected and used as starter cultures in the wet fermentation of pulped coffee. The MB and LAB populations varied from 4.48 to 8.43 log CFU g-1 for MB and 3.54 to 8.72 log CFU g-1 for LAB during fermentation. Organic acid concentration (ranged from 0.01 to 0.53 for succinic acid; 0.71 to 8.14 for lactic acid and 0.06 to 0.29 for acetic acid), and volatile compounds (44 compounds were detected in green beans and 98 in roasted beans) were evaluated during fermentation. The most abundant compounds found in roasted beans belong to furans [15], ketones and esters [14], pyridines [13], and pyrazines [12]). Leuconostoc mesenteroides CCMA 1105 and Lactobacillus plantarum CCMA 1065 presented volatile compounds important for coffee aroma. Isovaleric acid; 2,3-butanediol; phenethyl alcohol; ß-linalool; ethyl linoleate; and ethyl 2-hydroxypropanoate could improve cupping qualities.

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Volatile and non-volatile compounds in coffee directly affect the beverage's quality. This study aimed to demonstrate how the organic acids and volatile profiles were impacted by coffee fermentation using four starter cultures (Meyerozyma caribbica (CCMA 0198), Saccharomyces cerevisiae (CCMA 0543), Candida parapsilosis (CCMA0544), and Torulaspora delbrueckii (CCMA 0684)) inoculated in two varieties of coffee (Bourbon Amarelo and Canário Amarelo) using natural and pulped natural processing methods and sensory perception. Real-time PCR (qPCR) was used to verify the dynamic behavior of yeast populations. Organic acids were detected using high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) was used to detected volatile compounds. Sensory analysis was performed on the roasted coffee. Citric, malic, succinic, lactic, oxalic, isobutyric, and propionic acids and 105 volatile compounds were detected. At the beginning of fermentation, treatments with natural processing presented higher number of volatiles compounds. After fermentation, the main compounds groups were acids, alcohols, and aldehydes. The perception of sensory attribute (fruity, nutty, cocoa) varied with the coffee variety, type of processing, and type of inoculum. The use of yeasts is an alternative for sensorial differentiation of coffee variety Canário Amarelo and Bourbon Amarelo. The stainless-steel containers showed good results for coffee fermentation.

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This study evaluates the behavior of Saccharomyces (S.) cerevisiae (CCMA 0543), Candida (C.) parapsilosis (CCMA 0544), and Torulaspora (T.) delbrueckii (CCMA 0684) as starter cultures for semi-dry processed coffee using two inoculation methods: (1) direct inoculation and (2) bucket inoculation. The microbial population was evaluated by plating and real-time polymerase chain reaction (qPCR). The microbial metabolic changes of both bucket and direct inoculation methods during fermentation were evaluated using high performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC-MS). A sensorial test was also carried out. Citric and succinic acids were detected throughout the fermentation period. Chlorogenic acid concentration levels were higher for the bucket method after roasting. Roasted coffee beans also had a higher caffeine concentration, with the exception of the T. delbrueckii (CCMA0684) assay. Acids, pyrazines and pyridines were the main volatile compounds in both green and roasted coffee beans. Coffee cupping results proved that both inoculation methods scored well in terms of coffee quality. The bucket method favored the permanence of the microorganisms during coffee processing, especially the treatment inoculated with S. cerevisiae.

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