RESUMO
Traditional cocoa bean fermentation is a spontaneous process and can result in heterogeneous sensory quality. For this reason, yeast-integrated starter cultures may be an option for creating consistent organoleptic profiles. This study proposes the mixture of Hanseniaspora opuntiae and Kluyveromyces marxianus (from non-cocoa fermentation) as starter culture candidates. The microorganisms and volatile compounds were analyzed during the cocoa fermentation process, and the most abundant were correlated with predominant microorganisms. Results showed that Kluyveromyces marxianus, isolated from mezcal fermentation, was identified as the dominant yeast by high-throughput DNA sequencing. A total of 63 volatile compounds identified by HS-SPME-GC-MS were correlated with the more abundant bacteria and yeast using Principal Component Analysis and Agglomerative Hierarchical Clustering. This study demonstrates that yeasts from other fermentative processes can be used as starter cultures in cocoa fermentation and lead to the formation of more aromatic esters, decrease the acetic acid content.
Assuntos
Cacau , Fermentação , Hanseniaspora , Kluyveromyces , Compostos Orgânicos Voláteis , Compostos Orgânicos Voláteis/análise , Compostos Orgânicos Voláteis/metabolismo , Kluyveromyces/metabolismo , Hanseniaspora/metabolismo , Cacau/microbiologia , Cacau/metabolismo , Cacau/química , Microbiologia de Alimentos , Cromatografia Gasosa-Espectrometria de Massas , Ácido Acético/metabolismo , Fatores de TempoRESUMO
In addition to their pigment properties, the potential health benefits of anthocyanins have made them a subject of interest in recent years. This study aimed to obtain purified anthocyanin fractions from native Mexican black bean cultivars using Amberlite XAD-7 resin column and HPCCC and evaluate their anti-inflammatory properties using RAW 264.7 cells. The major anthocyanins in the purified anthocyanin fractions were delphinidin 3-glucoside (61.8%), petunidin 3-glucoside (25.2%), and malvidin 3-glucoside (12.2%). Purified anthocyanin fractions at 12.5 µg/mL effectively prevented LPS-induced ERK1/ERK2 phosphorylation and reduced the protein expression of COX-2 and mRNA expression of iNOS. Results showed that purified anthocyanin fractions have the potential to modulate the inflammatory response by inhibiting the production of pro-inflammatory mediators through the ERK1/ERK2 and NF-κB pathways. This study suggests that anthocyanins from black beans could be used as a natural strategy to help modulate inflammation-associated diseases.
Assuntos
Antocianinas , Anti-Inflamatórios , NF-kappa B , Extratos Vegetais , Antocianinas/farmacologia , Antocianinas/química , Antocianinas/isolamento & purificação , Camundongos , Células RAW 264.7 , Animais , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Extratos Vegetais/isolamento & purificação , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , NF-kappa B/genética , NF-kappa B/metabolismo , NF-kappa B/imunologia , Distribuição Contracorrente , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Inflamação/metabolismo , Inflamação/tratamento farmacológico , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Ciclo-Oxigenase 2/imunologia , Cromatografia Líquida de Alta Pressão , México , Phaseolus/química , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismoRESUMO
Cocoa bean fermentation is an important process because during this process, aroma compounds are produced, the astringency decreases, and the embryo dies. The fermentation processes of the Criollo and Forastero types have been studied separately without comparing them at the same time and in the same place. The aim of this work was to determine differences in the profile of volatile and nonvolatile compounds of Criollo and Forastero cocoa from the fermentation process to the final stage of obtaining the liquor. The experiments were carried out at the same time in the Maya region. Volatile compounds were determined by HS-SPME GC-MS (headspace solid phase-microextraction with gas chromatography-mass spectrometry). Sugars, organic acids, and alkaloids were determined by ultrahigh-performance liquid chromatography (UHPLC-PDA/UV). Criollo cocoa liquor was defined by the volatile and nonvolatile compounds such as acetic acid, phenylethyl alcohol, benzaldehyde, 2-phenylethyl acetate, acetophenone and 3-methylbutanal., which are associated with sour, honey, almond, flowery and chocolate aroma. Forastero cocoa liquor was represented with a significant difference by acetic acid, isobutyl acetate, 2,3-diethyl-5-methylpyrazine and ethyl octanoate and these could provide aroma descriptors such as sour, fruity and nutty. This study characterized for the first time the dynamics of volatile compounds during the fermentation, drying, and roasting stages and in the final cocoa liquor of Criollo and Forastero from cocoa beans of the same origin.
RESUMO
The isolation of autochthonous yeast species presents a good strategy to select new microorganisms for developing an adequate inoculum to carry out fermentations and generate representative products of the cider production zone. However, non-Saccharomyces yeasts have been considered to have low capacity to carry out a complete fermentation as Saccharomyces cerevisiae. In this work, five autochthonous yeasts from a cider fermentation process were isolated and identified as Saccharomyces cerevisiae, Kluyveromyces marxianus, Pichia membranaefaciens, P. kluyveri and Zygosaccharomyces rouxii. A series of fermentations were developed at laboratory level, using each species individually and it was observed that only S. cerevisiae was able to finish the process. K. marxianus consumed less than 50% of the sugars; P. kluyveri and Z. rouxii consumed less than 70% and P. membranaefaciens consumed more than 90% but the yield (ethanol produced for sugar consumed (YP/S)) was 0.39. Nevertheless, the addition of magnesium, zinc and nitrogen increased the fermentative capacity of almost all species: K. marxianus, Z. rouxii and P. kluyveri, showed an increase in ethanol production when nutrients were added, obtaining more than 80 g/L of ethanol, and showing that those nutrients are necessary to complete the fermentation. This work describes the potential use of different non-Saccharomyces species to carry out fermentation of apple juice and highlights the importance of certain nutrients to enable an efficient alcoholic fermentation and the generation of desirable volatile compounds for cider production.
Assuntos
Bebidas Alcoólicas/microbiologia , Nutrientes/metabolismo , Leveduras/metabolismo , Bebidas Alcoólicas/análise , Etanol/análise , Etanol/metabolismo , Fermentação , Sucos de Frutas e Vegetais/análise , Sucos de Frutas e Vegetais/microbiologia , Malus/microbiologia , Nutrientes/análise , Especificidade da Espécie , Leveduras/classificação , Leveduras/isolamento & purificaçãoRESUMO
Fermented cocoa beans can be described as a complex matrix that integrates the chemical history of beans, their processing, and environmental factors. This study presents an analysis that aims to identify volatile compounds of five varieties of fine-aroma cocoa types. The cocoa types studied were Carmelo, Rojo Samuel, Lagarto, Arcoiris, Regalo de Dios, that grow in the Maya lands of Chiapas, Mexico. Profile of volatile compounds was obtained from each cacao type during fermentation and drying process. This profile of volatile compounds also was compared with beans unfermented, using a statistical analysis of Venn diagram and a multivariate Analysis of Principal Components (PCA). One hundred nine different compounds were identified by SPME-HS GC-MS, these compounds mainly related to desirable aromatic notes generated by esters, aldehydes, ketones, and alcohols. The differences in chemical composition of the volatile compounds were associated mainly with the process and not to cocoa varieties. Fermented dry cocoa beans showed a higher content of esters, aldehydes, pyrazines, alcohols, some acids, and furans where Lagarto (CL), Rojo Samuel (CR), and Regalo de Dios (TRD) cocoas type showed a more interesting aromatic profile. On the other hand, as expected dry unfermented cocoas presented a few numbers of aroma compounds, in the five cacao types, where alcohols, ketones and hydrocarbons predominated.
Assuntos
Cacau/química , Fermentação , Manipulação de Alimentos , Análise de Componente Principal , Compostos Orgânicos Voláteis/análise , Álcoois/análise , Aldeídos/análise , Dessecação , Ésteres/análise , Análise de Alimentos , Cromatografia Gasosa-Espectrometria de Massas , Cetonas/análise , México , Odorantes/análiseRESUMO
Vinasses from the tequila industry are wastewaters with highly elevated organic loads. Therefore, to obtain value-added products by yeast fermentations, such as 2-phenylethanol (2-PE) and 2-phenylethylacetate (2-PEA), could be interesting for industrial applications from tequila vinasses. In this study, four yeasts species (Wickerhamomyces anomalus, Candida glabrata, Candida utilis, and Candida parapsilosis) were evaluated with two different chemically defined media and tequila vinasses. Differences in the aroma compounds production were observed depending on the medium and yeast species used. In tequila vinasses, the highest concentration (65 mg/L) of 2-PEA was reached by C. glabrata, the inhibitory compounds decreased biomass production and synthesis of 2-PEA, and biochemical and chemical oxygen demands were reduced by more than 50 %. Tequila vinasses were suitable for the production of 2-phenylethylacetate by the shikimate pathway. A metabolic network was developed to obtain a guideline to improve 2-PE and 2-PEA production using flux balance analysis (FBA).
RESUMO
Vinasse is a waste obtained from the production of beverages, such as tequila and cachaça. The presence of acids, alcohols, sugars, minerals, amino acids, peptides, and nitrogen salts make vinasse a hazardous liquid waste to the environment, affecting the fauna, flora, and microbiota of rivers and lagoons. This study used biological treatment concomitant to volatile compound production. The yeasts used in the study were Saccharomyces cerevisiae (CCMA 0187 and CCMA 0188), Candida parapsilosis (CCMA 0544), and Pichia anomala (CCMA 0193). A higher percentage reduction in chemical and biochemical oxygen demand was observed in the tequila vinasse than in the cachaça vinasse. However, a higher production of volatile compounds was observed in the cachaça vinasse. C. parapsilosis CCMA 0544 produced the highest concentration of 2-phenylethanol (162 mg L-1). These results indicated that the environmental damage of vinasse can be reduced by treating vinasse with yeasts, and this treatment produces aroma compounds. This biological treatment has high economic potential, especially for the tequila industry.
Assuntos
Bebidas Alcoólicas , Aromatizantes/metabolismo , Resíduos Industriais , Compostos Orgânicos Voláteis/metabolismo , Gerenciamento de Resíduos/métodos , Leveduras/metabolismo , Agave/química , Agave/microbiologia , Álcoois/metabolismo , Análise da Demanda Biológica de Oxigênio , Biomassa , Candida/metabolismo , Poluição Ambiental/prevenção & controle , Fermentação , Concentração de Íons de Hidrogênio , Álcool Feniletílico/metabolismo , Pichia/metabolismo , Saccharomyces cerevisiae/metabolismo , Saccharum , Temperatura , Compostos Orgânicos Voláteis/análise , Leveduras/crescimento & desenvolvimentoRESUMO
Enzymatic fructosylation of organic acceptors other than sugar opens access to the production of new molecules that do not exist in nature. These new glycoconjugates may have improved physical-chemical and bioactive properties like solubility, stability, bioavailability, and bioactivity. This review focuses on different classes of acceptors including alkyl alcohols, aromatic alcohols, alkaloids, flavonoids, and xanthonoids, which were tested for the production of fructoderivatives using enzymes from the glycoside hydrolase (GH) families 32 and 68 that use sucrose as donor substrate. The enzymatic strategies and the reaction conditions required for the achievement of these complex reactions are discussed, in particular with regard to the type of acceptors. The solubility and pharmacokinetic and antioxidant activity of some of these new ß-D-fructofuranosides in comparison is reviewed and compared with their glucoside analogs to highlight the differences between these molecules for technological applications.
Assuntos
Produtos Biológicos/metabolismo , Frutose/metabolismo , Glicosídeo Hidrolases/metabolismo , Sacarose/metabolismo , Produtos Biológicos/química , Glicosilação , Hexosiltransferases/metabolismo , Especificidade por SubstratoRESUMO
Agave tequilana fructans (ATF) constitute a substrate for bioethanol and tequila industries. As Kluyveromyces marxianus produces specific fructanases for ATF hydrolysis, as well as ethanol, it can perform simultaneous saccharification and fermentation. In this work, fifteen K. marxianus yeasts were evaluated to develop inoculums with fructanase activity on ATF. These inoculums were added to an ATF medium for simultaneous saccharification and fermentation. All the yeasts, showed exo-fructanhydrolase activity with different substrate specificities. The yeast with highest fructanase activity in the inoculums showed the lowest ethanol production level (20 g/l). Five K. marxianus strains were the most suitable for the simultaneous saccharification and fermentation of ATF. The volatile compounds composition was evaluated at the end of fermentation, and a high diversity was observed between yeasts, nevertheless all of them produced high levels of isobutyl alcohol. The simultaneous saccharification and fermentation of ATF with K. marxianus strains has potential for industrial application.
Assuntos
Agave/química , Biocombustíveis , Etanol/química , Glicosídeo Hidrolases/química , Kluyveromyces/metabolismo , Bebidas Alcoólicas , Biomassa , Butanóis/química , Carboidratos , Meios de Cultura , Fermentação , Hidrólise , Fatores de TempoRESUMO
Fructanase and fructosyltransferase are interesting for the tequila process and prebiotics production (functional food industry). In this study, one hundred thirty non-Saccharomyces yeasts isolated from "Mezcal de Oaxaca" were screened for fructanase and fructosyltransferase activity. On solid medium, fifty isolates grew on Agave tequilana fructans (ATF), inulin or levan. In liquid media, inulin and ATF induced fructanase activities of between 0.02 and 0.27U/ml depending of yeast isolate. High fructanase activity on sucrose was observed for Kluyveromyces marxianus and Torulaspora delbrueckii, while the highest fructanase activity on inulin and ATF was observed for Issatchenkia orientalis, Cryptococcus albidus, and Candida apicola. Zygosaccharomyces bisporus and Candida boidinii had a high hydrolytic activity on levan. Sixteen yeasts belonging to K. marxianus, T. delbrueckii and C. apicola species were positive for fructosyltransferase activity. Mezcal microbiota proved to showed to be a source for new fructanase and fructosyltransferases with potential application in the tequila and food industry.
Assuntos
Agave/enzimologia , Fermentação , Glicosídeo Hidrolases/metabolismo , Hexosiltransferases/metabolismo , Leveduras/enzimologia , Meios de CulturaRESUMO
An efficient method for the direct extraction of yeast genomic DNA from agave must was developed. The optimized protocol, which was based on silica-adsorption of DNA on microcolumns, included an enzymatic cell wall degradation step followed by prolonged lysis with hot detergent. The resulting extracts were suitable templates for subsequent qPCR assays that quantified mixed yeast populations in artisan Mexican mezcal fermentations.
Assuntos
Agave/microbiologia , DNA Fúngico/genética , Microbiologia de Alimentos/métodos , Leveduras/genética , Fermentação , México , Reação em Cadeia da PolimeraseRESUMO
A fructanase, produced by a Kluyveromyces marxianus strain isolated during the fermentation step of the elaboration process of "Mezcal de Guerrero" was purified and biochemically characterized. The active protein was a glycosylated dimer with a molecular weight of approximately 250 kDa. The specific enzymatic activity of the protein was determined for different substrates: sucrose, inulin, Agave tequilana fructan, levan and Actilight® and compared with the activity of Fructozyme®. The hydrolysis profile of the different substrates analyzed by HPAEC-PAD showed that the enzyme has different affinities over the substrates tested with a sucrose/inulin enzymatic activity ratio (S/I) of 125. For the hydrolysis of Agave tequilana fructans, the enzyme also showed a higher enzymatic activity and specificity than Fructozyme®, which is important for its potential application in the tequila industry.