Whasley Ferreira Duarte

Fermentative behavior of Saccharomyces strains during microvinification of raspberry juice (Rubus idaeus L.)

Sixteen different strains of Saccharomyces cerevisiae and Saccharomyces bayanus were evaluated in the production of raspberry fruit wine. Raspberry juice sugar concentrations were adjusted to 16° Brix with a sucrose solution, and batch fermentations were performed at 22 °C. Various kinetic parameters, such as the conversion factors of the substrates into ethanol (Y(p/s)), biomass (Y(x/s)), glycerol (Y(g/s)) and acetic acid (Y(ac/s)), the volumetric productivity of ethanol (Q(p)), the biomass productivity (P(x)), and the fermentation efficiency (E(f)) were calculated. Volatile compounds (alcohols, ethyl esters, acetates of higher alcohols and volatile fatty acids) were determined by gas chromatography (GC-FID). The highest values for the E(f), Y(p/s), Y(g/s), and Y(x/s) parameters were obtained when strains commonly used in the fuel ethanol industry (S. cerevisiae PE-2, BG, SA, CAT-1, and VR-1) were used to ferment raspberry juice. S. cerevisiae strain UFLA FW 15, isolated from fruit, displayed similar results. Twenty-one volatile compounds were identified in raspberry wines. The highest concentrations of total volatile compounds were found in wines produced with S. cerevisiae strains UFLA FW 15 (87,435 μg/L), CAT-1 (80,317.01 μg/L), VR-1 (67,573.99 μg/L) and S. bayanus CBS 1505 (71,660.32 μg/L). The highest concentrations of ethyl esters were 454.33 μg/L, 440.33 μg/L and 438 μg/L for S. cerevisiae strains UFLA FW 15, VR-1 and BG, respectively. Similar to concentrations of ethyl esters, the highest concentrations of acetates (1927.67 μg/L) and higher alcohols (83,996.33 μg/L) were produced in raspberry wine from S. cerevisiae UFLA FW 15. The maximum concentration of volatile fatty acids was found in raspberry wine produced by S. cerevisiae strain VR-1. We conclude that S. cerevisiae strain UFLA FW 15 fermented raspberry juice and produced a fruit wine with low concentrations of acids and high concentrations of acetates, higher alcohols and ethyl esters.

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.

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.

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.

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.

Bacterial dynamics and chemical changes during the spontaneous production of the fermented porridge (Calugi) from cassava and corn

-1 and were the predominant group; acetic acid bacteria were also observed at the onset of fermentation (3.82 log CFU ml -1 ). The population of lactic acid bacteria was 3.7 log CFU ml -1 and remained constant throughout the fermentation. The denaturing gradient gel electrophoresis (DGGE) bands were identified as Corynebacterium variabile, Lactobacillus paracasei, L. plantarum, L. casei, Bacillus spp. (Bacillus cereus group), B. subtilis, Streptomyces sp., Enterobacter cloacae, Streptococcus parasanguis, Streptococcus salivarius, Weissella cibaria and Weissella confusa, in addition to uncultivable bacteria. The most abundant carbohydrate in the substrate was maltose (38 g L -1 at the beginning of fermentation). Acetic and lactic acids were the principal acids found by high

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.

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.

Biotechnological applications of coffee processing by-products

The search for new substrates to be used in biotechnological processes has grown in recent years, especially, for the bioprocesses in which microorganisms are used to produce value-added products. In this scenario, the coffee processing by-products, wastewater, pulp, mucilage, husk, silverskin, and spent coffee ground, appear as interesting alternative substrate to be used in bacteria, filamentous fungi, and yeast cultivation. The composition of the coffee processing by-products including water-soluble sugars, proteins, lignin, cellulose, hemicellulose, oils, etc., allows its microbial conversion into products, such as ethanol, acids (lactic, citric, gibberellic, and gallic), aromatic volatile compounds (terpenes, higher alcohols, and esters), pigments (carotenoids), microbial biomass (edible mushroom), and enzymes. In this context, the current chapter presents several examples of coffee processing by-products used as substrate for the cultivation of microorganisms with an ultimate goal to generate value-added products.