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This study aims to characterize the physicochemical properties of cocoa’s residual honey and evaluate its fermentative capacity as a substrate, using Saccharomyces cerevisiae AWRI726 as the starter culture for alcoholic fermentation. The research hypothesis was that cocoa’s residual honey can be used for the production of fermented beverages. Cocoa’s honey has 14.14 g.100 mL−1 of dry material, containing 11.80 g.100 mL−1 of carbohydrates and 1.20% crude protein, in addition to other minor components, such as pectin, lipids, and Fe, Mn, Na, and Zn, with a carbon-to-nitrogen (C/N) ratio (9.8) most suitable for fermentation. Fermentation at 20°C for 240 hours produced a liquid with 16% v/v ethanol (14 g.L−1 in 144 h). However, 24 hours of fermentation produced the maximum ethanol yield (0.373 g.g−1) and volumetric productivity (0.168 g.L−1.h−1), which were associated with a significant increase in the specific cell growth rate. Saccharomyces cerevisiae AWR1726 performed satisfactorily in the alcoholic fermentation of cocoa’s residual honey, similar to that observed in other fruit beverages, thus suggesting the suitability of cocoa’s residual honey for future commercial applications.

Cell immobilization comprises the retention of metabolically active cells inside a polymeric matrix. In this study, the production of jabuticaba (Myrciaria jaboticaba) vinegar using immobilized Acetobacter aceti and Gluconobacter oxydans cells is proposed as a new method to prevent losses of jabuticaba fruit surplus. The pulp of jabuticaba was processed and Saccharomyces cerevisiae CCMA 0200 was used to ferment the must for jabuticaba wine production. Sugars, alcohols (ethanol and glycerol) and organic acids were assayed by high-performance liquid chromatography. Volatile compounds were determined by gas chromatography-flame ionization detector. The ethanol content of the produced jabuticaba wine was approx. 74.8 g/L (9.5 % by volume) after 168 h of fermentation. Acetic acid fermentation for vinegar production was performed using a mixed culture of immobilized A. aceti CCT 0190 and G. oxydans CCMA 0350 cells. The acetic acid yield was 74.4 % and productivity was 0.29 g/(L·h). The vinegar had particularly high concentrations of citric (6.67 g/L), malic (7.02 g/L) and succinic (5.60 g/L) acids. These organic acids give a suitable taste and flavour to the vinegar. Seventeen compounds (aldehydes, higher alcohols, terpene, acetate, diether, furans, acids, ketones and ethyl esters) were identified in the jabuticaba vinegar. In conclusion, vinegar was successfully produced from jabuticaba fruits using yeast and immobilized mixed cultures of A. aceti and G. oxydans. To the best of our knowledge, this is the first study to use mixed culture of immobilized cells for the production of jabuticaba vinegar.

The aim of this study was to develop a kefir apple-based vinegar and evaluate this fermentation process using new methodology with Biospeckle Laser. Brazilian kefir grains were inoculated in apple must for vinegar production. In this study, the microbial community present in kefir, and correspondent vinegar, was investigated using Matrix Assisted Laser Desorption/Ionization - Time of Flight Mass Spectrometry (MALDI-TOF MS) technique. Saccharomyces cerevisiae, Lactobacillus paracasei, Lactobacillus plantarum, Acetobacter pasteurianus and Acetobacter syzygii were the microbial species identified. S. cerevisiae, L. plantarum, A. pasteurianus and A. syzygii were found in smaller quantities at the beginning of the alcoholic fermentation, but were found throughout the alcoholic and acetic fermentation. Kefir grains were able to utilize apple must as substrate to produce ethanol, and acetic acid. Acetate, volatile alcohols and aldehydes in the vinegar-based kefir were also produced. The yield of acetic acid in the kefir vinegars was ∼79%. The acetic acid concentration was ∼41 g L-1, reaching the required standard for the Brazilian legislation accepts it as vinegar (4.0% acetic acid). Kefir vinegar showed good acceptance in the sensory analysis. The technology proposed here is novel by the application of immobilized-cell biomass (kefir grains) providing a mixed inocula and eliminating the use of centrifuge at the end of the fermentative process. This step will save energy demand and investment. This is the first study to produce apple vinegar using kefir grains.

Biochemical and molecular analysis was used for identification of different kefir yeasts species from Brazil, Canada and the United States of America. The sugar/ethanol-resistant activity of the yeasts was evaluated. Saccharomyces cerevisiae and Kluyveromyces marxianus had the highest growth rates, suggesting biotechnological applications possible for these strains.

Cell immobilization comprises the retention of metabolically active cells inside a polymeric matrix. In this study, the production of jabuticaba (Myrciaria jaboticaba) vinegar using immobilized Acetobacter aceti and Gluconobacter oxydans cells is proposed as a new method to prevent losses of jabuticaba fruit surplus. The pulp of jabuticaba was processed and Saccharomyces cerevisiae CCMA 0200 was used to ferment the must for jabuticaba wine production. Sugars, alcohols (ethanol and glycerol) and organic acids were assayed by high-performance liquid chromatography. Volatile compounds were determined by gas chromatography-flame ionization detector. The ethanol content of the produced jabuticaba wine was approx. 74.8 g/L (9.5 % by volume) after 168 h of fermentation. Acetic acid fermentation for vinegar production was performed using a mixed culture of immobilized A. aceti CCT 0190 and G. oxydans CCMA 0350 cells. The acetic acid yield was 74.4 % and productivity was 0.29 g/(L·h). The vinegar had particularly high concentrations of citric (6.67 g/L), malic (7.02 g/L) and succinic (5.60 g/L) acids. These organic acids give a suitable taste and flavour to the vinegar. Seventeen compounds (aldehydes, higher alcohols, terpene, acetate, diether, furans, acids, ketones and ethyl esters) were identified in the jabuticaba vinegar. In conclusion, vinegar was successfully produced from jabuticaba fruits using yeast and immobilized mixed cultures of A. aceti and G. oxydans. To the best of our knowledge, this is the first study to use mixed culture of immobilized cells for the production of jabuticaba vinegar., Imobilizacija stanica podrazumijeva zadržavanje metabolički aktivnih stanica unutar polimernog matriksa. U radu je predložena nova metoda proizvodnje octa iz plodova biljke Myrciaria jaboticaba pomoću imobiliziranih stanica bakterija Acetobacter aceti i Gluconobacter oxydans, za sprečavanje gubitaka u proizvodnji tog voća zbog viška plodova. Fermentacijom voćne pulpe pomoću kvasca Saccharomyces cerevisiae CCMA 0200 nastao je mošt, koji je zatim upotrijebljen za proizvodnju vina. Koncentracije šećera, alkohola (etanola i glicerola) i organskih kiselina u vinu određene su visokodjelotvornom tekućinskom kromatografijom, a koncentracije hlapljivih spojeva plinskom kromatografijom s plameno- -ionizacijskim detektorom. Koncentracija etanola u proizvedenom vinu bila je otprilike 74,8 g/L (volumni udjel 9,5 %) nakon 168 sati fermentacije. Pomoću mješovite kulture imobilizoranih stanica bakterija A. aceti CCT 0190 i G. oxydans CCM 0350 proizveden je ocat. Dobiven je prinos octene kiseline od 74,4 %, a produktivnost je postupka iznosila 0,29 g/(L·h). Proizvedeni ocat imao je izrazito velike koncentracije limunske (6,67 g/L), jabučne (7,02 g/L) i sukcinske kiseline (5,60 g/L). Te su mu organske kiseline dale prikladan okus i aromu. Identificirano je 17 različitih spojeva (aldehida, viših alkohola, terpena, acetata, dietera, furana, kiselina, ketona i etilnih estera) u octu. Zaključeno je da je ocat uspješno proizveden iz plodova biljke Myrciaria jaboticaba pomoću kvasca i imobilizirane mješovite kulture A. aceti i G. oxydans. Prema našim saznanjima, ovo je prvo istraživanje u kojem su za proizvodnju octa iz plodova biljke Myrciaria jaboticaba upotrebljene imobilizirane stanice mješovite kulture bakterija.

Bacaba chicha is a beverage prepared by the indigenous Umutina people from the bacaba fruit (Oenocarpus bacaba), a purple berry that is rich in fat and carbohydrates, as well as a source of phenolic compounds. In this study, samples of bacaba chicha beverage were collected, and the microbial community was assessed using culture-dependent and -independent techniques. The nutritional composition and metabolite profiles were analyzed, and species belonging to lactic acid bacteria (LAB) and yeasts were detected. The LAB group detected by culture-dependent analysis included Enterococcus hormaechei and Leuconostoc lactis. Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) detected additional Propionibacterium avidum, Acetobacter spp., and uncultured bacteria. Pichia caribbica and Pichia guilliermondii were detected in a culture-dependent method, and Pichia caribbica was confirmed by PCR-DGGE analysis. The pH value of the beverage was 6.2. The nutritional composition was as follows: 16.47 ± 0.73 g 100 mL-1 dry matter, 2.2 ± 0.0 g 100 mL-1 fat, 3.36 ± 0.44 g 100 mL-1 protein, and 10.87 ± 0.26 g 100 mL-1 carbohydrate. The metabolites detected were 2.69 g L-1 succinic acid, 0.9 g L-1 acetic acid, 0.49 g L-1 citric acid, 0.52 g L-1 ethanol, and 0.4 g L-1 glycerol. This is the first study to identify microbial diversity in bacaba chicha spontaneous fermentation. This study is also the starting step in the immaterial record of this Brazilian indigenous beverage prepared from bacaba fruit.

A total of 2152 Actinobacteria strains were isolated from native Cerrado (Brazilian Savannah) soils located in Passos, Luminárias, and Arcos municipalities (Minas Gerais State, Brazil). The soils were characterised for chemical and microbiological analysis. The microbial analysis led to the identification of nine genera (Streptomyces, Arthrobacter, Rhodococcus, Amycolatopsis, Microbacterium, Frankia, Leifsonia, Nakamurella,andKitasatospora) and 92 distinct species in both seasons studied (rainy and dry). The rainy season produced a high microbial population of all the aforementioned genera. The pH values of the soil samples from the Passos, Luminárias, and Arcos regions varied from 4.1 to 5.5. There were no significant differences in the concentrations of phosphorus, magnesium, and organic matter in the soils among the studied areas. Samples from the Arcos area contained large amounts of aluminium in the rainy season and both hydrogen and aluminium in the rainy and dry seasons. The Actinobacteria population seemed to be unaffected by the high levels of aluminium in the soil. Studies are being conducted to produce bioactive compounds from Actinobacteria fermentations on different substrates. The present data suggest that the number and diversity of Actinobacteria spp. in tropical soils represent a vast unexplored resource for the biotechnology of bioactives production.

Microbial ecology and chemical composition of Brazilian kefir beverage was performed. The microorganisms associated with Brazilian kefir were investigated using a combination of phenotypic and genotypic methods. A total of 359 microbial isolates were identified. Lactic acid bacteria (60.5%) were the major isolated group identified, followed by yeasts (30.6%) and acetic acid bacteria (8.9%). Lactobacillus paracasei (89 isolates), Lactobacillus parabuchneri (41 isolates), Lactobacillus casei (32 isolates), Lactobacillus kefiri (31 isolates), Lactococcus lactis (24 isolates), Acetobacter lovaniensis (32 isolates), Kluyveromyces lactis (31 isolates), Kazachstania aerobia (23 isolates), Saccharomyces cerevisiae (41 isolates) and Lachancea meyersii (15 isolates) were the microbial species isolated. Scanning electron microscopy showed that the microbiota was dominated by bacilli (short and curved long) cells growing in close association with lemon-shaped yeasts cells. During the 24 h of fermentation, the protein content increased, while lactose and fat content decreased. The concentration of lactic acid ranged from 1.4 to 17.4 mg/ml, and that of acetic acid increased from 2.1 to 2.73 mg/ml. The production of ethanol was limited, reaching a final mean value of 0.5 mg/ml., The authors acknowledge Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES) for scholarshipsand and CAARG - Cooperativa Agricola Alto Rio Grande Ltda. (Lavras/MG, Brazil) for the milk supply.

Whey valorization concerns have led to recent interest on the production of whey beverage simulating kefir. In this study, the structure and microbiota of Brazilian kefir grains and beverages obtained from milk and whole/deproteinised whey was characterized using microscopy and molecular techniques. The aim was to evaluate its stability and possible shift of probiotic bacteria to the beverages. Fluorescence staining in combination with Confocal Laser Scanning Microscopy showed distribution of yeasts in macro-clusters among the grain’s matrix essentially composed of polysaccharides (kefiran) and bacteria. Denaturing gradient gel electrophoresis displayed communities included yeast affiliated to Kluyveromyces marxianus, Saccharomyces cerevisiae, Kazachatania unispora, bacteria affiliated to Lactobacillus kefiranofaciens subsp. Kefirgranum, Lactobacillus kefiranofaciens subsp. Kefiranofaciens and an uncultured bacterium also related to the genus Lactobacillus. A steady structure and dominant microbiota, including probiotic bacteria, was detected in the analyzed kefir beverages and grains. This robustness is determinant for future implementation of whey-based kefir beverages., Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), CAPES-GRICES.

Cheese whey (CW) and deproteinised cheese whey (DCW) were investigated for their suitability as novel substrates for the production of kefir-like beverages. Lactose consumption, ethanol production, as well as organic acids and volatile compounds formation, were determined during CW and DCW fermentation by kefir grains and compared with values obtained during the production of traditional milk kefir. The results showed that kefir grains were able to utilise lactose from CW and DCW and produce similar amounts of ethanol (7.8–8.3 g/l), lactic acid (5.0 g/l) and acetic acid (0.7 g/l) to those obtained during milk fermentation. In addition, the concentration of higher alcohols (2-methyl-1-butanol, 3-methyl-1-butanol, 1-hexanol, 2-methyl-1-propanol, and 1-propanol), ester (ethyl acetate) and aldehyde (acetaldehyde) in cheese whey-based kefir and milk kefir beverages were also produced in similar amounts. Cheese whey and deproteinised cheese whey may therefore serve as substrates for the production of kefir-like beverages similar to milk kefir., The authors acknowledge the financial support from Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES), CAPES-GRICES and Lactogal for supplying cheese whey powder.