Arcopilus eremanthusum sp. nov. as sources of antibacterial and antioxidant metabolites.

In this study, we evaluated the antibacterial and antioxidant activities of ethyl acetate (EtOAc) extracts of Arcopilus eremanthusum sp. nov. (CML3766) isolated from E. erythopappus. The fungi were identified using the sequences of the internal transcribed spacer (ITS), large subunit (LSU), and RNA polymerase II second largest subunit (RPB2). Antibacterial activity was determined using the minimum bactericidal concentration (MBC) method. Antioxidant activity was evaluated using free radical scavenging methods with 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS•+) assays and the ß-carotene-linoleic acid system. The total phenolic compound content was measured using the Folin-Ciocalteu method. The endophytic fungal extract presented bactericidal activity, with an MBC of 2.44 and 19.5 µg/mL against Staphylococcus aureus GL 8702 and GL 5674, respectively, and 625 µg/mL for Salmonella enterica serovar Enteritidis. In addition, this fungus demonstrated an antioxidant activity of 52.30% protection in the ß-carotene method. The total concentration of phenolic compounds was 23.73 mg gallic acid equivalent (GAE)/g. Ferulic acid, trans-cinnamic acid, chlorogenic acid, catechin, vanillin, p-coumaric acid, and caffeic acid were quantified using high-performance liquid chromatography with diode array detection. The results demonstrate the potential of A. eremanthusum sp. nov. to serve as a source of antibacterial and antioxidant metabolites with possible future biotechnological applications.

Production and characterization of a new distilled beverage from green coffee seed residue.

This study evaluated green coffee seed residue (GCSR) as an alternative substrate for producing distilled beverages. Two proportions of GCSR, 10% and 20% (w/v), were fermented and distilled in a copper alembic still. The spirits were characterized by GC-FID, HS-SPME GC-MS, and sensory analysis by trained panelists. Most of the 62 identified volatile compounds were affected by the GCSR concentration. Total terpenes, higher alcohols, and acetals showed the highest concentrations in the 10% GCSR spirit. Esters, acetates, and aldehydes were most abundant in the 20% GCSR. In the sensory analysis, the 10% GCSR spirit was characterized by floral, dairy, and almond aromas, while the 20% GCSR spirit was embodied coffee, vegetable, hazelnut, cooked cabbage, and nut descriptors. The results demonstrate the potential of GCSR as a substrate for producing coffee spirits with chemical and sensory qualities, with the 10% GCSR being the better option for fermentation.

Production and characterization of a new distillate obtained from fermentation of wet processing coffee by-products.

Coffee is one of the most important commodities worldwide. The industrial processing of coffee cherries generates a considerable volume of by-products such as wastewater, coffee pulp, mucilage, and husk. These by-products have sugars and nutrients that can be converted into value-added products via microbial action. In this study, for the first time, we evaluated the potential of coffee pulp and coffee wastewater as substrate for alcoholic fermentation produce a distilled beverage. The must composed by dry or wet coffee pulp and coffee wastewater added of commercial sucrose or sugarcane molasses was fermented by S. cerevisiae. After a screening step, a larger fermentation was carried out with the wet pulp added of sucrose due to its higher alcoholic fermentation efficiency. The distilled beverage contained 38% (v/v) ethanol and 0.2 g/L of acetic acid. The contaminants furfural, hydroxymethylfurfural and ethyl carbamate were below detection level. Among the 48 volatile compounds detected, the majority (21) were ethyl esters usually associated with floral and sweet aromas. Ethyl decanoate (996.88 µg/L) and ethyl dodecanoate (1088.09 µg/L) were the most abundant esters. Coffee spirit presented taste acceptance of 80% and sugarcane spirit, 70%. The tasters indicated an aroma acceptance of 86% for the coffee spirit and 78% for the sugarcane spirit. The results of this work demonstrate the potential for using coffee by-products to produce a good quality distilled beverage. Considering our results, especially sensorial analysis, we can infer that the produced coffee beverage represents a new alternative for adding value to the coffee production chain.

HS-SPME/GC-MS Assisted Analysis of Volatile Constituents in Different Strains of Shiitake Culinary- Medicinal Mushroom, Lentinus edodes (Agaricomycetes).

The shiitake culinary-medicinal mushroom Lentinus edodes is one of the most consumed species worldwide because it has nutritional, medicinal, and palatable properties. Its organoleptic characteristics contribute substantially to its high popularity. The pleasant aromas result from the presence of volatile compounds. The objective of the present work was to study the profiles of volatile constituents of fresh fruiting bodies of five strains of L. edodes. The volatile compounds were extracted by solid phase microextraction method and analyzed by gas chromatography and mass spectrometry. The aromatic profiles of the strains revealed variability. Both alcohols and sulfides were the most abundant volatile compounds. LE6 strain presented the highest number of volatile compounds with predominance of sulfides (dimethyl pentasulfide, s-tetrathiane) and for LE2, the aldehydes were the most representative chemical class with the main volatile being (E)-2-octen-1-al.

Survival of Kluyveromyces lactis and Torulaspora delbrueckii to simulated gastrointestinal conditions and their use as single and mixed inoculum for cheese production.

The demand for new probiotic products has shown recent increases alongside a growing interest in studying starter cultures of cheeses. This study thus aims to evaluate the ability to survive under simulated gastrointestinal conditions and impact of Torulaspora delbrueckii B14 and Kluyveromyces lactis B10 as single and mixed inocula for cheese production. These two yeast strains were subjected to simulated gastrointestinal tracts and tested for self-aggregation, hydrophobicity, pathogen inhibition, antibiotic resistance, and ß-galactosidase production. The yeast strains were also assessed for their ability to survive in different NaCl concentrations (2.5%, 5%, and 10% w/v), multiple temperatures (4 °C and 40 °C), and used as single and mixed starter cultures for cheese production. Yeasts population levels were monitored by YPD plating and MALDI-TOF and metabolites were analyzed by HPLC and GC-MS over the course of the 21 days cheese maturation process. T. delbrueckii B14 and K. lactis B10 both showed >80% viability after the passage through the simulated gastrointestinal tract, had self-aggregation rates >90%, and displayed ß-galactosidase activities of 0.35 U/g and 0.53 U/g, respectively. Both yeasts survived at 2.5%, 5%, and 10% NaCl for 21 days and showed growth at 4 °C. In cheese, the single inoculum of K. lactis B10 and mixed inoculum showed the highest levels of lactose consumption. HS-SPME GC-MS analysis of cheese samples allowed the identification of 38 volatile compounds. The highest concentrations of most of these compounds were observed after 21 days of maturation for the cheese produced with mixed inoculum. The most abundant acids detected were hexanoic and decanoic acid; the most abundant alcohols were 2,3-butanediol, 2-phenylethanol and isoamyl alcohol, and the most prevalent ester compounds were isoamyl acetate and phenethyl acetate. Our results therefore show that T. delbrueckii B14 and K. lactis B10 are interesting yeasts for further studies in the context of probiotics and positively impact the composition of desirable volatile compounds in cheeses, particularly when used as mixed inoculum.

Probiotic potential of yeasts isolated from pineapple and their use in the elaboration of potentially functional fermented beverages.

Pineapple (Ananas comosus (L.) Merril) is a tropical fruit rich in nutrients characterized by a pleasant taste and widely consumed in several countries. It is used to produce juice, jams and wine. In this work, 150 yeasts isolates were obtained from peel (18) and spontaneously fermented pineapple pulp (132). The probiotic potential of 50 isolates was studied. Survival at pH 2.0, pepsin 3.0 g/L, and tolerance of bile salts (0.1 and 1% (w/v) were determined as indicators of survival potential of the isolates during the passage through the human gastrointestinal tract in simulated conditions. The selected isolates were also evaluated for their resistance to 6 antibiotics, antimicrobial activity against 6 pathogenic bacteria and autoaggregation and hydrophobicity properties. Five of them survived to gastrointestinal conditions, showed antibiotic resistance and autoaggregation properties. They were identified by MALDI-TOF MS and sequencing of ITS region as Candida lusitaniae (3) and Meyerozyma caribbica (2). Among these isolates, M. caribbica 9 D was evaluated in the production of a fermented pineapple beverage. The Saccharomyces cerevisiae var. boulardii was used as control, due to the fact that it is the only commercially available probiotic yeast. With M. caribbica inoculum, the beverage produced showed higher concentrations of residual glucose (24.19 g/L) and fructose (8.67 g/L), lower concentration of acetic acid (0.22 g/L); higher total phenolic compounds (196.93 mg/L), catechin (155.56 mg/L), chlorogenic acid (3.64 mg/L), vanillin (0.18 mg/L) and ferulic acid (33.2 mg/L). It was observed that M. caribbica population remained stable during refrigerated storage with cell counts greater than 7.00 log CFU/mL over 21 days. Compared to beverage produced with S. cerevisiae var. boulardii, the one produced with M. caribbica presented greater acceptance in the sensorial analysis for taste, aroma and general acceptance. The fermented pineapple beverage prepared with M. caribbica proved to be a good alternative in development of a potential probiotic beverage with different sensory and nutritional properties.

Yeasts from Canastra cheese production process: Isolation and evaluation of their potential for cheese whey fermentation.

Canastra cheese is a cheese with geographical indication recognized by the Brazilian National Institute of Industrial Protection under number IG201002. It is produced in seven municipalities in the state of Minas Gerais in a region called Serra da Canastra. In this work, samples of milk, "pingo" (natural starter), whey and Canastra cheese were collected on a farm in Medeiros-MG/Brazil to evaluate the yeast microbiota and select yeasts for whey fermentation to produce ethanol and volatile aromatic compounds of relevance in the production of cheese. Thirty-nine isolates capable of fermenting lactose in a synthetic medium were identified by MALDI-TOF as Kluyveromyces lactis (29), Torulaspora delbrueckii (7) and Candida intermedia (3). Eleven isolates of K. lactis and three of T. delbrueckii efficiently fermented lactose until 4th day, and due to this reason were selected for cheese whey fermentation with Brix 12, 14 and 18. Generally, the isolates T. delbrueckii B14, B35, and B20 and K. lactis B10 were the most effective regardless of the initial Brix value. The identification of these four isolates by MALDI TOF was confirmed by sequencing of the ITS region. In the fermentation of cheese whey 14 Brix, T. delbrueckii B14 and B35, respectively yielded 24.06g/L and 16.45g/L of ethanol, while K. lactis B10 was more efficient in the consumption of lactose. In sequential culture with K. lactis B10 inoculated 48h after T. delbrueckii B14, 97.82% of the total sugars were consumed resulting in the production of 19.81g/L ethanol and 39 aromatic volatile compounds. The most abundant compounds were 3-methyl-1-butanol, octanoic acid and ethyl decanoate, which are reported as important for the aroma and flavor of cheeses. Based in our results, B10 isolate inoculated 48h after B14 isolate is a promising yeast inoculum to be used for fermentation of dairy substrates.

Parâmetros microbiológicos em diferentes cortes de carne bovina resfriada / Microbiological parameters in different cuts of cooled beef

O objetivo deste estudo foi verificar a contagem de mesófilos, coliformes totais, Staphylococcus aureus, Escherichia coli e presença de Salmonella sp. em cortes de traseiro de bovinos obtidos em um abatedouro frigorífico sob Inspeção Federal. Os resultados indicaram que todos os cortes mostraram-se ausentes para Salmonella sp. e a maior média de contagem para mesófilos foi no Filé de costela, seguido pela Picanha, Coxão mole, Fralda, Pera (recorte do Coxão Mole) e Maminha. Destes, a Pera e a Picanha apresentaram os maiores valores para Staphylococcus aureus. As maiores médias de contagens em UFC/g para coliformes totais foram nos cortes: Coxão mole, Alcatra, Pera e Filé-Mignon. De forma geral, os níveis de coliformes totais foram baixos, porém revelaram a presença de Escherichia coli principalmente no Coxão Mole. A Alcatra apresentou resultados elevados para coliformes totais, porém, sem a presença de E. coli. Os resultados dos cortes coletados mostraram que não houve desvio aparente nos processos quanto às operações do abate, a desossa e refile.

The aim of this study was to verify the mesophilic count, total coliforms, Staphylococcus aureus, Escherichia coli and the presence of Salmonella sp. in cattle rear cuts obtained in a slaughterhouse governed under Federal Inspection. The results indicated that all the cuts proved to be absent for Salmonella sp. and the highest average count for mesophilic was in rib steak, followed by Top Sirloin Cap, Topside, Flank Steak, Cut of Topside and Tenderloin. Of these, Cut of Topside and Top Sirloin Cap showed the highest values for S. aureus. The highest average scores in CFU/g for total coliforms were in cuts: Topside, Rump Steak, Cut of Topside and Tenderloin. The levels of total coliforms were low and showed the presence of Escherichia coli in the Topside. The results showed high thighs of coliforms, but without the presence of E. coli. The results of the collected cuttings showed no apparent shift in processes and operations during slaughter and boning and trimming.

Sugar cane spirit (cachaça): Effects of mixed inoculum of yeasts on the sensory and chemical characteristics.

The main goal of this study was to produce cachaça using a mixed inoculum of Saccharomyces cerevisiae and Meyerozyma caribbica and characterize the produced beverage using HPLC, GC-FID, GC-MS and sensorial analysis. Additionally, the use of MALDI-TOF as a tool to characterize and monitor pure and mixed inocula fermenting sugar cane juice was also evaluated. Vat fermentations were carried out for three consecutive batches using autoclaved 16°Brix sugar cane juice fermented by a mixed inoculum of M. caribbica 107 cells/mL and S. cerevisiae 108 cells/mL. The cachaça produced by the mixed culture of M. caribbica and S. cerevisiae showed the highest concentration of volatile compounds associated with good sensory descriptors such as ethyl hexanoate (114.11µg/L), 2-phenylethyl acetate (2.77µg/L), a-terpineol (0.45µg/L), b-citronellol (2.47µg/L), and geraniol (0.24µg/L). This beverage consequently showed greater acceptance in the sensorial analysis for taste and aroma, especially by younger panelists. The feasibility of MALDI-TOF use under studied conditions was demonstrated by the comparison of the results obtained from yeast cultivation in YPD broth, YPD agar and sugar cane juice, showing that there was no interference of sugar cane juice in protein profile. The results obtained from MALDI-TOF analysis showed that the protein extraction directly from sugar cane juice under fermentation, without the traditional plating step, allowed the distinction between mixed and pure inocula even under different M. caribbica populations and Brix degrees.

In vitro determination of volatile compound development during starter culture-controlled fermentation of Cucurbitaceae cotyledons.

The effects of Lactobacillus plantarum UFLA CH3, Pediococcus acidilactici UFLA BFFCX 27.1, and Torulaspora delbrueckii UFLA FFT2.4 inoculation on the volatile compound profile of fermentation of Cucumeropsis mannii cotyledons were investigated. Different microbial associations were used as starters. All associations displayed the ability to ferment the cotyledons as judged by lowering the pH from 6.4 to 4.4-5 within 24h and increasing organic acids such as lactate and acetate. The population of lactic acid bacteria (LAB) and yeasts increased during fermentation. In the fermentation performed without inoculation (control), the LAB and yeast populations were lower than those in inoculated assays at the beginning, but they reached similar populations after 48 h. The Enterobacteriaceae population decreased during the fermentation, and they were not detected at 48 h in the L. plantarum UFLA CH3 and P. acidilactici UFLA BFFCX 27.1 (LP+PA) and L. plantarum UFLA CH3, P. acidilactici UFLA BFFCX 27.1, and T. delbrueckii UFLA FFT2.4 (LP+PA+TD) samples. The assays inoculated with the yeast T. delbrueckii UFLA FFT2.4 exhibited the majority of volatile compounds (13 compounds) characterized by pleasant notes. The LP+PA+TD association seemed to be appropriate to ferment C. mannii cotyledons. It was able to control the Enterobacteriaceae population, and achieved high concentrations of esters and low concentrations of aldehydes and ketones.

Utilization of coffee by-products obtained from semi-washed process for production of value-added compounds.

The semi-dry processing of coffee generates significant amounts of coffee pulp and wastewater. This study evaluated the production of bioethanol and volatile compounds of eight yeast strains cultivated in a mixture of these residues. Hanseniaspora uvarum UFLA CAF76 showed the best fermentation performance; hence it was selected to evaluate different culture medium compositions and inoculum size. The best results were obtained with 12% w/v of coffee pulp, 1 g/L of yeast extract and 0.3 g/L of inoculum. Using these conditions, fermentation in 1 L of medium was carried out, achieving higher ethanol yield, productivity and efficiency with values of 0.48 g/g, 0.55 g/L h and 94.11% respectively. Twenty-one volatile compounds corresponding to higher alcohols, acetates, terpenes, aldehydes and volatile acids were identified by GC-FID. Such results indicate that coffee residues show an excellent potential as substrates for production of value-added compounds. H. uvarum demonstrated high fermentative capacity using these residues.

Study of the physicochemical parameters and spontaneous fermentation during the traditional production of yakupa, an indigenous beverage produced by Brazilian Amerindians.

Yakupa is a traditional non-alcoholic cassava beverage produced by Brazilian Amerindians. In this work the microbial dynamics and metabolites involved in yakupa fermentation were investigated by PCR-denaturing gradient gel electrophoresis and chromatography analysis, respectively. The lactic acid bacteria (LAB) population was higher than yeast in the beginning of fermentation (5 log CFU mL(-1) and 3 log CFU mL(-1), respectively) and after 36 h both population increased reaching 7 log CFU mL(-1), remaining constant until 60 h. Culture dependent and independent methods in combination identified the bacteria Lactobacillus fermentum, L. plantarum, Weissela cibaria and W. confusa, and yeasts Saccharomyces cerevisiae and Pichia kudriavzevii. Maltose (41.2 g L(-1)), ethanol (6.5 g L(-1)) and lactic acid (7.8 g L(-1)) were the most abundant compounds identified by high performance liquid chromatography. Aldehydes, acids, alcohols and esters were identified by gas chromatography flame ionization detection. By the metabolites and PCA analysis we may assign the beverage's flavor to the microbial metabolism. Heterolactic LAB and S. cerevisiae dominated the yakupa fermentation, being responsible for the organoleptic characteristics of the final product. This is the first time that the microbial dynamics and physicochemical parameters were investigated in the yakupa beverage and it may contribute to the future selection of starter cultures to perform yakupa fermentations.

Evaluation of adhesive properties of presumptive probiotic Lactobacillus plantarum strains / Avaliação das propriedades de adesão de presumíveis estirpes probióticas de Lactobacillus plantarum

Thirty-two strains of Lactobacillus plantarum UFLA SAU from pork sausages, pre-selected for some features for probiotic application, were utilized in this study to evaluate their adhesive properties and compare the results against the three pathogens also tested. Strains were tested for autoaggregation and coaggregation capacity and Microbial Adhesion To Solvents (MATS) at the time intervals of 0, 1, 2, 3 and 4 h. Our findings revealed that UFLA SAU strains have a high autoaggregative capacity and coaggregative ability with pathogens, especially Listeria monocytogenes. In relation to adhesion to solvents, in general, L. plantarum strains showed hydrophilic cell surface properties and an important electron donor and basic character. Adhesive properties were markedly separated for the strains under study by Principal Component Analysis software. UFLA SAU 132, 226 and 87 were differentiated by autoaggregation ability. UFLA SAU 11 and Listeria monocytogenes were characterized by adhesion to solvents. UFLA SAU 14, 18 and 172 showed high coaggregation with Escherichia coli, Salmonella Typhi and Listeria monocytogenes. In comparison to the pathogens tested, many UFLA SAU strains presented higher adhesive capacity. These tests should be used for screening and identifying potentially adherent microorganisms. Adhesive properties are important features for the choice of probiotic strains and confer various applications, such as in the pharmaceutical (therapeutic or prophylactic) and food (functional foods) industries.

Trinta e duas estirpes de linguiça suína, Lactobacillus plantarum UFLA SAU, pré-selecionadas com algumas características para aplicação probiótica, foram utilizadas neste estudo para avaliar suas propriedades adesivas e comparar os resultados com três patógenos também testados. As estirpes foram testadas para autoagregação, coagregação e capacidade de adesão microbiana aos solventes (MATS) nos tempos de 0, 1, 2, 3 e 4 h. Nossos resultados revelaram que estirpes UFLA SAU apresentam alta capacidade autoagregativa e coagregativa com patógenos, especialmente com Listeria monocytogenes. Em relação à adesão aos solventes, de um modo geral, as estirpes de L. plantarum mostraram propriedades hidrofílicas de superfície celular e um importante caráter básico e elétron doador. Propriedades adesivas foram marcadamente separadas para as estirpes em estudo através da Análise de Componentes Principais. UFLA SAU 132, 226 e 87 foram diferenciadas pela capacidade de autoagregação. UFLA SAU 11 e Listeria monocytogenes foram caracterizadas por adesão aos solventes. UFLA SAU 14, 18 e 172 apresentaram coagregação com Escherichia coli, Salmonella Typhi e Listeria monocytogenes. Em comparação aos patógenos testados, muitas estirpes UFLA SAU apresentaram maior capacidade adesiva. Estes testes podem ser úteis para a triagem e identificação de micro-organismos potencialmente aderentes. Propriedades adesivas são importantes características para a escolha de estirpes probióticas e conferem várias aplicações, tais como nas indústrias: farmacêutica (terapêutico ou profilático) e de alimentos (alimentos funcionais).

Evaluation of stress tolerance and fermentative behavior of indigenous Saccharomyces cerevisiae

Sixty six indigenous Saccharomyces cerevisiae strains were evaluated in stressful conditions (temperature, osmolarity, sulphite and ethanol tolerance) and also ability to flocculate. Eighteen strains showed tolerant characteristics to these stressful conditions, growing at 42 ºC, in 0.04% sulphite, 1 mol L-1 NaCl and 12% ethanol. No flocculent characteristics were observed. These strains were evaluated according to their fermentative performance in sugar cane juice. The conversion factors of substrates into ethanol (Yp/s), glycerol (Yg/s) and acetic acid (Yac/s), were calculated. The highest values of Yp/s in sugar cane juice fermentation were obtained by four strains, one isolated from fruit (0.46) and the others from sugar cane (0.45, 0.44 and 0.43). These values were higher than the value obtained using traditional yeast (0.38) currently employed in the Brazilian bioethanol industry. The parameters Yg/s and Yac/s were low for all strains. The UFLA FW221 presented the higher values for parameter related to bioethanol production. Thus, it was tested in co-culture with Lactobacillus fermentum. Besides this, a 20-L vessel for five consecutive batches of fermentation was performed. This strain was genetically stable and remained viable during all batches, producing high amounts of ethanol. The UFLA FW221 isolated from fruit was suitable to produce bioethanol in sugar cane juice. Therefore, the study of the biodiversity of yeasts from different environmental can reveal strains with desired characteristics to industrial applications.

Screening of Lactobacillus isolated from pork sausages for potential probiotic use and evaluation of the microbiological safety of fermented products.

The aim of this study was to select strains of Lactobacillus isolated from pork sausage for use as probiotics. Lactobacillus isolates were evaluated in tests based on probiotic characteristics and microbiological safety. The UFLA SAU 14, 52, and 91 isolates were differentiated by coaggregation with Listeria monocytogenes, production of lactic acid, and survival at pH 2. UFLA SAU 172 and 187 isolates had high levels of coaggregation with Salmonella Typhi and Escherichia coli, tolerance to pancreatic fluid, and adhesion to chloroform. UFLA SAU 20 and 34 isolates were characterized by exopolysaccharide production, autoaggregation, and resistance to simulated intestinal fluid. UFLA SAU 185, 238, and 258 isolates exhibited resistance to bile and adhesion to xylene. A cocktail of these 10 Lactobacillus isolates with potential probiotic properties was inoculated into pork sausage and inhibited the growth of L. monocytogenes.

The effects of co-culturing non-Saccharomyces yeasts with S. cerevisiae on the sugar cane spirit (cachaça) fermentation process.

Twenty non-Saccharomyces strains were previously tested in pure culture for their ability to grow in 12 % ethanol, their ß-glucosidase activity, flocculation, glycerol, ethanol and acetic acid production, fermentation kinetics and their production of volatile compounds. Of these 20 strains, three strains, namely, Pichia anomala UFLA CAF70, P. anomala UFLA CAF119 and Pichia caribbica UFLA CAF733, were evaluated in co-culture with Saccharomyces cerevisiae UFLA CA11. Of the mixed inocula, the mixture of P. caribbica UFLA CAF733 and S. cerevisiae UFLA CA11 gave the highest ethanol concentration (75.37 g/L), the lowest levels of residual glucose (1.14 g/L) and fructose (19.92 g/L), and the highest volumetric productivity (Q (p)) of ethanol. Twenty-three minor volatile compounds were identified in the fermented sugar cane juice. The mixed culture of P. caribbica UFLA CAF733 and S. cerevisiae UFLA CA11 gave the highest concentration of volatile compounds with good sensory descriptors; these compounds included ethyl esters (290.13 µg/L), acetates (715.21 µg/L) and monoterpenic alcohols (195.56 µg/L). This mixed culture also gave the lowest concentration of volatile acids (1774.46 µg/L) and aldehydes (121.10 µg/L). In principal component analysis, the mixed inoculum of UFLA CAF733 and UFLA CA11 was positively characterized by ethyl hexanoate, 2-phenylethanol, linalool, nonanoic acid, ethyl butyrate, phenylethyl acetate, diethylsuccinate, hexanoic acid, and geraniol. In conclusion, we found that clear improvements could be achieved in the fermentation process with mixed, rather than pure, S. cerevisiae culture. The use of the non-Saccharomyces strain P. caribbica UFLA CAF733 in co-culture with S. cerevisiae UFLA CA11 may therefore be an interesting means by which to improve the quality of cachaça.

Evaluation of a potential starter culture for enhance quality of coffee fermentation.

The coffee fermentation is characterized by the presence of different microorganisms belonging to the groups of bacteria, fungi and yeast. The objectives of this work were to select pectinolytic microorganisms isolated from coffee fermentations and evaluate their performance on coffee pulp culture medium. The yeasts and bacteria isolates were evaluated for their activity of polygalacturonase (PG), pectin lyase (PL) and pectin methylesterase (PME) and metabolites production. Among 127 yeasts isolates and 189 bacterial isolates, 15 were pre-selected based on their ability to produce PL and organic compounds. These isolates were strains identified as Bacillus cereus, Bacillus megaterium, Bacillus subtilis, Candida parapsilosis, Pichia caribbica, Pichia guilliermondii and Saccharomyces cerevisiae. When cultivated in Coffee peel and pulp media in single culture or two by two mixed inocula, different behavior concerning to PME, PL and PG were found. The two principal components PC1 and PC2 accounted for 45.27 and 32.02 % of the total variance. UFLA CN727 and UFLA CN731 strains were grouped in the positive part of PC1 being characterized by 1,2-propanediol, hexanoic acid, decanoic acid, nonanoic acid and ethyl acetate. The UFLA CN448 and UFLA CN724 strains were grouped in the negative part of PC1 and were mainly characterized by guaiacol, butyric acid and citronellol. S. cerevisiae UFLACN727, P. guilliermondii UFLACN731 and C. parapsilosis UFLACN448 isolates are promising candidates to be tested in future studies as coffee starter cultures.

Evaluation of stress tolerance and fermentative behavior of indigenous Saccharomyces cerevisiae.

Sixty six indigenous Saccharomyces cerevisiae strains were evaluated in stressful conditions (temperature, osmolarity, sulphite and ethanol tolerance) and also ability to flocculate. Eighteen strains showed tolerant characteristics to these stressful conditions, growing at 42 °C, in 0.04% sulphite, 1 mol L(-1) NaCl and 12% ethanol. No flocculent characteristics were observed. These strains were evaluated according to their fermentative performance in sugar cane juice. The conversion factors of substrates into ethanol (Y(p/s)), glycerol (Y(g/s)) and acetic acid (Y(ac/s)), were calculated. The highest values of Y(p/s) in sugar cane juice fermentation were obtained by four strains, one isolated from fruit (0.46) and the others from sugar cane (0.45, 0.44 and 0.43). These values were higher than the value obtained using traditional yeast (0.38) currently employed in the Brazilian bioethanol industry. The parameters Y(g/s) and Y(ac/s) were low for all strains. The UFLA FW221 presented the higher values for parameter related to bioethanol production. Thus, it was tested in co-culture with Lactobacillus fermentum. Besides this, a 20-L vessel for five consecutive batches of fermentation was performed. This strain was genetically stable and remained viable during all batches, producing high amounts of ethanol. The UFLA FW221 isolated from fruit was suitable to produce bioethanol in sugar cane juice. Therefore, the study of the biodiversity of yeasts from different environmental can reveal strains with desired characteristics to industrial applications.

Effect of co-inoculation of Saccharomyces cerevisiae and Lactobacillus fermentum on the quality of the distilled sugar cane beverage cachaça.

UNLABELLED: The effect of Lactobacillus fermentum in co-inoculation with Saccharomyces cerevisiae UFLA CA11 on the quality of cachaça (sugar cane spirit) was evaluated. The co-inoculation was first evaluated in flask fermentation, and subsequently, yeast and bacteria were co-cultured at approximately 10(5) CFU/mL and 10(8) CFU/mL, respectively, in 4 consecutive batches. L. fermentum did not affect the growth or activity (sugar consumption and fermentation rate) of S. cerevisiae UFLA CA11 during fermentation. The physicochemical analysis revealed a higher concentration (Tukey test) of aldehydes (22.07 mg/100 mL anhydrous alcohol) in cachaça produced by co-inoculation. Analysis of volatile compounds using GC-FID demonstrated that cachaça produced by co-inoculation had higher concentrations of acetaldehyde (25.15 mg/L), ethyl acetate (30.17 mg/L), and 2,3-butanedione (170.39 µ/L), while cachaça produced by UFLA CA11 contained higher concentrations of ethyl lactate (1205.98 µ/L), propionic acid (127.97 µ/L), butyric acid (2335.57 µ/L), and 1-pentanol (469.23 µ/L). The lowest concentration of acetic acid measured by HPLC was found in cachaça obtained with UFLA CA11. The sensory analysis, performed using the Mann-Whitney test, revealed that cachaça produced by co-inoculation differed from that produced by UFLA CA11 in taste and aroma. PRACTICAL APPLICATION: This study reports on the use of a mixed culture of Saccharomyces cerevisae and Lactobacillus fermentum to produce cachaça and shows the influence of co-inoculation of yeast and bacteria on the quality of this beverage.

Optimization of fermentation conditions for production of the jabuticaba (Myrciaria cauliflora) spirit using the response surface methodology.

UNLABELLED: The jabuticaba tree (Brazilian grape tree) is a tree native to Brazil that belongs to the Myrtaceae family. The jabuticaba fruit is used in some regions of Brazil to produce juices, jams, wine, and ice cream. In this work, the fermentation conditions (temperature and °Brix) for producing jabuticaba distillate were optimized using the response surface methodology (RSM). The optimal conditions for fermentation were found to be 20 °C and 22 °Brix. In repeated experiments to validate the model, experimental data exhibited good agreement with the predicted data. The distillate jabuticaba beverage showed a peculiar chemical composition with 20 volatile compounds that were identified and quantified. Isoamyl alcohols (2-methyl-1-butanol + 3-methyl-1-butanol) were the most abundant volatile compounds identified in jabuticaba spirit. Sensory analysis by tasters showed overall approval of jabuticaba distillate. In principal component analysis, when the beverage was evaluated by panelists under 24 y old tended to give favorable ratings of aroma and taste, as well as high overall scores. The group of panelists between the ages of 25 and 53 y old generally gave high marks for appearance in the principal component analysis. PRACTICAL APPLICATION: This study describes the study of fermentation conditions of jabuticaba pulp for production of jabuticaba spirit. Based on the results of this work, the proposed method can be an alternative for the use of the jabuticaba fruit, and may provide a new industrial outlet for this fruit.