[In silico analysis of Cit s 2: A highly conserved profilin]. / Análisis in silico de cit s 2: una profilina altamente conservada.

OBJECTIVE: Analyze phylogenetic relationships and molecular mimicry of Cit s 2 and other plant profilins. METHODS: Online bioinformatics tools including Basic Local Alignment Search Tool (BLASTP), PRALINE and MEGA were used for multiple alignments and phylogenetic analysis. A 3D-homology model of Cit s 2 was predicted. Models were calculated with MODELLER. The best model was selected with the model scoring option of MAESTRO. Conserved regions between Cit s 2 and other profilins were located on the 3D model and antigenic regions were predicted by ElliPro server (3-5). RESULTS: Cit s 2 amino acid sequence (Uniprot code:P84177) was compared with other 30 profilins from different allergenic sources. The identity between Cit s 2 and other profilins ranged between 82 and 99%. The highest identity was observed with Cucumis melo (99%) followed by Prunus persica (98%) and Malus domestica (92%). High conserved antigenic regions were observed on the 3D predicted model. Seven lineal and six discontinuous epitopes were found in Cit s 2. CONCLUSION: High conserved antigenic regions were observed on the 3D predicted model of Cit s 2, which might involve potential cross-reactivity between Cit s 2 and other profilins. Future studies are needed to further analyze these results.

OBJETIVO: Analizar las relaciones filogenéticas y el mimetismo molecular de Cit s 2 y otras profilinas vegetales. MÉTODOS: Se utilizaron herramientas bioinformáticas en línea, incluida la de búsqueda de alineación local básica (BLASTP), PRALINE y MEGA, para alineamientos múltiples y análisis filogenético. Se predijo un modelo de homología 3D de Cit s 2. Los modelos se calcularon con MODELLER. El mejor modelo fue seleccionado con la opción de puntuación de modelo de Maestro. Las regiones conservadas entre Cit s 2 y otras profilinas se ubicaron en el modelo 3D y las regiones antigénicas fueron predichas por el servidor ElliPro (3-5). RESULTADOS: La secuencia de aminoácidos de Cit s 2 (código Uniprot: P84177), se comparó con otras 30 profilinas de diferentes fuentes alergénicas. La mayor identidad se observó con Cucumis melo (99%) seguida de Prunus persica (98%) y Malus domestica (92%). Se observaron regiones antigénicas altamente conservadas en el modelo predicho en 3D. Se encontraron siete epítopes lineales, y seis epítopes discontinuos en Cit s 2. CONCLUSIÓN: Se observaron regiones antigénicas altamente conservadas en el modelo 3D predicho de Cit s 2, lo que podría implicar una posible reactividad cruzada entre Cit s 2 y otras profilinas. Se necesitan estudios futuros para analizar más a fondo estos resultados.

Superficial scald development in 'Granny Smith' and 'Nicoter' apples: The role of key volatile compounds when fruit are stored under dynamic controlled atmosphere.

A positive correlation of α-farnesene and its oxidation metabolites with superficial scald is commonly reported in apples stored in air or controlled atmosphere (CA) systems, where O2 levels are above the lower oxygen limit (LOL) tolerated by the fruit. Nevertheless, the LOL can be monitored by the dynamic controlled atmosphere (DCA) techniques and to provide different physiological responses. Therefore, this study aimed to evaluate key volatile metabolites from 'Granny Smith' and 'Nicoter' ('Kanzi®') apples stored under dynamic controlled atmosphere (DCA) monitored by respiratory quotient (RQ), i. e. at extremely low oxygen partial pressures (ELO) and correlate their emissions with the incidence of superficial scald (SS). The volatile compounds (VCs) were isolated by solid phase microextraction (HS-SPME) and analyzed by gas chromatography. For the first time, higher concentrations of α-farnesene and its oxidation metabolites (6-methyl-5-hepten-2-one, and 6-methyl-5-hepten-2-ol) were negatively correlated with SS in DCA-RQ. This is likely due to the higher levels of ethanol in fruit stored under this condition having an inhibitory effect on SS incidence even when α-farnesene, 6-methyl-5-hepten-2-one, and 6-methyl-5-hepten-2-ol accumulate. Additionally, SS is more correlated to internal ethylene concentration (IEC) than α-farnesene accumulation and their metabolites, even when fruit were stored under ELO, where ethylene action is reduced.

Superabsorbent bacterial cellulose film produced from industrial residue of cashew apple juice processing.

The industrial residue of cashew apple juice processing (MRC) was evaluated as an alternative medium for bacterial cellulose (BC) production by Komagataeibacter xylinus ATCC 53582 and Komagataeibacter xylinus ARS B42. The synthetic Hestrin-Schramm medium (MHS) was used as a control for growing and BC production. First, BC production was assessed after 4, 6, 8, 10, and 12 days under static culture. After 12 days of cultivation, K. xylinus ATCC 53582 produced the highest BC titer in MHS (3.1 g·L-1) and MRC (3 g·L-1), while significant productivity was attained at 6 days of fermentation. To understand the effect of culture medium and fermentation time on the properties of the obtained films, BC produced at 4, 6, or 8 days were submitted to infrared spectroscopy with Fourier transform, thermogravimetry, mechanical tests, water absorption capacity, scanning electron microscopy, degree of polymerization and X-ray diffraction. The properties of BC synthesized in MRC were identical to those of BC from MHS, according to structural, physical, and thermal studies. MRC, on the other hand, allows the production of BC with a high water absorption capacity when compared to MHS. Despite the lower titer (0.88 g·L-1) achieved in MRC, the BC from K. xylinus ARS B42 presented a high thermal resistance and a remarkable absorption capacity (14664 %), suggesting that it might be used as a superabsorbent biomaterial.

Novel fungal organophosphorus hydrolases in acidic media: an application to apples decontamination.

Organophosphorus pesticides bring significant improvements in agriculture, but their toxicity causes environmental and health negative impacts. The aim of this work was the development of robust biocatalysts to be applied in bioremediation. Four fungi were evaluated as hydrolase sources capable of degrading organophosphorus pesticides: Aspergillus niger, Fusarium sp., Penicillium chrysogenum, and Penicillium nalgiovense. The hydrolysis rates of methyl paraoxon obtained under acidic conditions were in the range of 10 to 21 mg L-1 d-1, which is remarkable since most similar biocatalysts are active under alkaline conditions. Penicillium chrysogenum activity was outstanding, and it was selected to prepare, characterize, and study the applications of its enzymatic extract. It was used to evaluate the bioremediation of apple surfaces at pH 2 in the presence of SDS, achieving complete methyl paraoxon degradation under proposed conditions. These results indicate that this biocatalyst could complement industrialized fruit washing processes for the elimination of organophosphorus pesticides.

The roles of WRKY transcription factors in Malus spp. and Pyrus spp.

The WRKY transcription factor gene family is known to be involved in plant defense against pathogens and in tolerance to different environmental stresses at different stages of development. The response mechanisms through which these genes act can be influenced by different phytohormones as well as by many trans- and cis-acting elements, making this network an important topic for analysis, but still something complex to fully understand. According to available reports, these genes can also perform important roles in pome species (Malus spp. and Pyrus spp.) metabolism, especially in adaptation of these plants to stressful conditions. Here, we present a quick review of what is known about WRKY genes in Malus and Pyrus genomes offering a simple way to understand what is already known about this topic. We also add information connecting the evolution of these transcription factors with others that can also be found in pomes.

Structural Studies of the Cutin from Two Apple Varieties: Golden Delicious and Red Delicious (Malus domestica).

The cuticle, a protective cuticular barrier present in almost all primary aerial plant organs, has a composition that varies between plant species. As a part of the apple peel, cuticle and epicuticular waxes have an important role in the skin appearance and quality characteristic in fresh fruits destined for human consumption. The specific composition and structural characteristics of cutin from two apple varieties, "golden delicious" and "red delicious", were obtained by enzymatic protocols and studied by means of cross polarization magic angle spinning nuclear magnetic resonance (CP-MAS 13C NMR), attenuated total reflection infrared spectroscopy (ATR-FTIR), and mass spectrometry, and were morphologically characterized by specialized microscopy techniques (atomic force microscopy (AFM), confocal laser scanning microscopy (CLMS), and scanning electron microscopy (SEM)). According to CP-MAS 13C NMR and ATR-FTIR analysis, cutins from both varieties are mainly composed of aliphatics and a small difference is shown between them. This was corroborated from the hydrolyzed cutins analysis by mass spectrometry, where 9,10,18-trihydroxy-octadecanoic acid; 10,20-Dihydroxy-icosanoic acid; 10,16-dihydroxy hexadecenoic acid (10,16-DHPA); 9,10-epoxy-12-octadecenoic acid; and 9,10-epoxy-18-hydroxy-12-octadecenoic acid were the main monomers isolated. The low presence of polysaccharides and phenolics in the cutins obtained could be related to the low elastic behavior of this biocomposite and the presence of cracks in the apple cutin's surface. These cracks have an average depth of 1.57 µm ± 0.57 in the golden apple, and 1.77 µm ± 0.64 in those found in the red apple. The results obtained in this work may facilitate a better understanding that mechanical properties of the apple fruit skin are mainly related to the specific aliphatic composition of cutin and help to much better investigate the formation of microcracks, an important symptom of russet formation.

Valorization of apple pomace using bio-based technology for the production of xylitol and 2G ethanol.

Apple pomace was studied as a raw material for the production of xylitol and 2G ethanol, since this agroindustrial residue has a high concentration of carbohydrate macromolecules, but is still poorly studied for the production of fermentation bioproducts, such as polyols. The dry biomass was subjected to dilute-acid hydrolysis with H2SO4 to obtain the hemicellulosic hydrolysate, which was concentrated, detoxified and fermented. The hydrolyzate after characterization was submitted to submerged fermentations, which were carried out in Erlenmeyer flasks using, separately, the yeasts Candida guilliermondii and Kluyveromyces marxianus. High cellulose (32.62%) and hemicellulose (23.60%) contents were found in this biomass, and the chemical hydrolysis yielded appreciable quantities of fermentable sugars, especially xylose. Both yeasts were able to metabolize xylose, but Candida guilliermondii produced only xylitol (9.35 g L-1 in 96 h), while K. marxianus produced ethanol as the main product (10.47 g L-1 in 24 h) and xylitol as byproduct (9.10 g L-1 xylitol in 96 h). Maximum activities of xylose reductase and xylitol dehydrogenase were verified after 24 h of fermentation with C. guilliermondii (0.23 and 0.53 U/mgprot, respectively) and with K. marxianus (0.08 e 0.08 U/mgprot, respectively). Apple pomace has shown potential as a raw material for the fermentation process, and the development of a biotechnological platform for the integrated use of both the hemicellulosic and cellulosic fraction could add value to this residue and the apple production chain.

Technical benefit on apple fruit of controlled atmosphere influenced by 1-MCP at molecular levels.

The apple is a highly perishable fruit after harvesting and, therefore, several storage technologies have been studied to provide the consumer market with a quality product with a longer shelf life. However, little is known about the apple genome that is submitted to the storage, and even less with the application of ripening inhibitors. Due to these factors, this study sought to elucidate the transcriptional profile of apple cultivate Gala stored in a controlled atmosphere (AC) treated and not treated with 1-methyl cyclopropene (1-MCP). Through the genetic mapping of the apple, applying the microarray technique, it was possible to verify the action of treatments on transcripts related to photosynthesis, carbohydrate metabolism, response to hormonal stimuli, nucleic acid metabolism, reduction of oxidation, regulation of transcription and metabolism of cell wall and lipids. The results showed that the transcriptional profile in the entire genome of the fruit showed significant differences in the relative expression of the gene, this in response to CA in the presence and absence of 1-MCP. It should be noted that the transcription genes involved in the anabolic pathway were only maintained after six months in fruits treated with 1-MCP. The data in this work suggests that the apple in the absence of 1-MCP begins to prepare its metabolism to mature, even during the storage period in AC. Meanwhile, in the presence of the inhibitor, the transcriptional profile of the fruit is similar to that at the time of harvest. It was also found that a set of genes that code for ethylene receptors, auxin homeostasis, MADS Box, and NAC transcription factors may be involved in the regulation of post-harvest ripening after storage and in the absence of 1-MCP.

Functional characterisation and in silico modelling of MdPSY2 variants and MdPSY5 phytoene synthases from Malus domestica.

Carotenoids are plastid isoprenoid pigments that play critical roles in light harvesting, photoprotection, and phytohormone biosynthesis. They are also vitamin-A precursors and antioxidant molecules important for human nutrition. Apples (e.g. Malus x domestica Borkh), one of the most widely consumed fruits with high nutrient levels, have a very low carotenoid concentration in flesh, compared with other fruits and vegetables. This could be explained by a deficiency in carotenoid synthesis/accumulation and/or accelerated degradation. We analysed the contribution of M. domestica cv. 'Fuji' phytoene synthase (PSY) in the biosynthesis of carotenoids and determined that among four MdPSY genes present in the organism, MdPSY2 and MdPSY5 are highly expressed in leaves and during fruit ripening in line with an increment in carotenoid content in fruits. Furthermore, two representative polymorphic MdPSY2 variants were found, one with a Tyr358Phe substitution (MdPSY2_F) and the other that additionally has a six-amino-acid deletion in the signal peptide (MdPSY2_CG). MdPSY2, MdPSY5, MdPSY2_F and MdPSY2_CG are all localised in plastids. Interestingly, the polymorphic MdPSY2_F and MdPSY2_CG variants show lower enzymatic activity than the wild-type form in a heterologous complementation assay, which could be attributed to the Tyr358Phe substitution close to the active-site pocket, as was suggested by 3-D modelling analysis. The presence of polymorphic MdPSY2 variants with lower enzymatic activity could be partially responsible for the low carotenoid content in Fuji apple fruits.

Boosting carotenoid content in Malus domestica var. Fuji by expressing AtDXR through an Agrobacterium-mediated transformation method.

Apple (Malus domestica) fruits accumulate negligible levels of carotenoids, antioxidant pigments that are precursors for vitamin A in humans. As vitamin A deficiency is an important public health issue, we aimed at increasing carotenoids in apple by constitutively expressing the Arabidopsis thaliana DXR gene, one of the key regulatory steps in the plastidial isoprenoid pathway. For this purpose, we optimized an Agrobacterium-mediated transformation method in the commercial Fuji Raku Raku variety. This resulted in a shoot establishment efficiency of 0.75% at 20 weeks after infection. Molecular and microscopical analyses revealed that 80% of the hygromycin resistant shoots contained and expressed AtDXR:eGFP and that the AtDXR:eGFP fusion protein located in plastids. Transgenic seedlings displayed up to 3-fold increase in total carotenoids and in individual carotenoids compared to the WT, correlating with an increased transcript abundance of endogenous carotenogenic genes such as MdDXS, MdPSY1, MdPSY2, MdPSY3, MdLCYB1, and MdLCYB2. In addition, buds of 2-year-old transgenic dormant trees showed an increment up to 3-fold in lutein, and transient transformation of fruits revealed that AtDXR induced a 2-fold increment in total carotenoids. Thus, these results suggest that DXR may be a good candidate for increasing carotenoid levels in apple fruits through metabolic engineering.

Simultaneous extraction and separation of bioactive compounds from apple pomace using pressurized liquids coupled on-line with solid-phase extraction.

The objective of this work was the development of an on-line extraction/fractionation method based on the coupling of pressurized liquid extraction and solid-phase extraction for the separation of phenolic compounds from apple pomace. Several variables of the process were evaluated, including the amount of water of the first stage (0-120 mL), temperature (60-80 °C), solid-phase extraction adsorbent (Sepra, Isolute, Strata X and Oasis) and activation/elution solvent (methanol and ethanol). The best results were observed with the adsorbent Sepra. The temperature had a small effect on recovery, but significant differences were observed for phlorizin and a quercetin derivative. Results indicate that ethanol can be used to replace methanol as an activation, extraction/elution solvent. While using mostly green solvents (water, ethanol, and a small amount of methanol that could be reused), the developed method produced higher or similar yields of acids (2.85 ± 0.19 mg/g) and flavonoids (0.97 ± 0.11 mg/g) than conventional methods.

Apple bagasse as a substrate for the propagation of Patagonian wine yeast biomass.

AIMS: A culture medium based on apple bagasse was designed and tested as a substrate for biomass production of conventional and unconventional native wine yeasts. METHODS AND RESULTS: The physicochemical characterization of the apple bagasse was carried out and its potential utility as a constituent of a complete culture medium for the production of yeast biomass was analysed using the experimental statistical designs. Growth parameters of conventional and nonconventional Patagonian wine yeasts were analysed with Placket-Burman designs and response surface methodology, comparing in each assay the apple bagasse substrate with the commonly used substrate for biomass development, cane molasses. Culture media composition was optimized and models were validated. CONCLUSIONS: This study demonstrates that, both from a nutritional and from an economic point of view, apple bagasse constitutes a more advantageous substrate than cane molasses for the propagation of native yeasts from Patagonia. SIGNIFICANCE AND IMPACT OF THE STUDY: We used an alternate carbon-rich material, generously available in our region, originally generated as fruit industrial waste, to transform it into a source of sustainable, economically profitable and environmentally friendly energy resource.

High-throughput analysis of pesticides in minor tropical fruits from Colombia.

In this work, a method has been applied and validated for the determination of a group of 35 multiclass pesticides in the minor tropical fruits rose apple/pomarrosa (Syzygium malaccense), starfruit/carambola (Averrhoa carambola), yoyomo (Spondias purpurea) and papayuela (Vasconcellea pubescens) cultivated and exported in Colombia. The AOAC 2007.1 QuEChERS method, that uses an acetic acid/acetate buffer, was applied together with gas chromatography coupled to a triple quadrupole mass spectrometer. The method was validated in terms of calibration, recovery at three levels of concentration and matrix effects (MEs). A mixture of analyte protectants was also used. A good linearity was obtained in all cases, while the study of the ME revealed the need of developing matrix-matched calibration for many pesticides. Recovery values were in the range 70-120% with relative standard deviation values less than 20% for most of the pesticides studied. The lowest calibration level was 5 µg/kg. Several samples of each type were analysed.

Evolutionary diversification of galactinol synthases in Rosaceae: adaptive roles of galactinol and raffinose during apple bud dormancy.

Galactinol synthase (GolS) is a key enzyme in the biosynthetic pathway of raffinose family oligosaccharides (RFOs), which play roles in carbon storage, signal transduction, and osmoprotection. The present work assessed the evolutionary history of GolS genes across the Rosaceae using several bioinformatic tools. Apple (Malus × domestica) GolS genes were transcriptionally characterized during bud dormancy, in parallel with galactinol and raffinose measurements. Additionally, MdGolS2, a candidate to regulate seasonal galactinol and RFO content during apple bud dormancy, was functionally characterized in Arabidopsis. Evolutionary analyses revealed that whole genome duplications have driven GolS gene evolution and diversification in Rosaceae speciation. The strong purifying selection identified in duplicated GolS genes suggests that differential gene expression might define gene function better than protein structure. Interestingly, MdGolS2 was differentially expressed during bud dormancy, concomitantly with the highest galactinol and raffinose levels. One of the intrinsic adaptive features of bud dormancy is limited availability of free water; therefore, we generated transgenic Arabidopsis plants expressing MdGolS2. They showed higher galactinol and raffinose contents and increased tolerance to water deficit. Our results suggest that MdGolS2 is the major GolS responsible for RFO accumulation during apple dormancy, and these carbohydrates help to protect dormant buds against limited water supply.

Contrasting Transcriptional Programs Control Postharvest Development of Apples (Malus x domestica Borkh.) Submitted to Cold Storage and Ethylene Blockage.

Apple is commercially important worldwide. Favorable genomic contexts and postharvest technologies allow year-round availability. Although ripening is considered a unidirectional developmental process toward senescence, storage at low temperatures, alone or in combination with ethylene blockage, is effective in preserving apple properties. Quality traits and genome wide expression were integrated to investigate the mechanisms underlying postharvest changes. Development and conservation techniques were responsible for transcriptional reprogramming and distinct programs associated with quality traits. A large portion of the differentially regulated genes constitutes a program involved in ripening and senescence, whereas a smaller module consists of genes associated with reestablishment and maintenance of juvenile traits after harvest. Ethylene inhibition was associated with a reversal of ripening by transcriptional induction of anabolic pathways. Our results demonstrate that the blockage of ethylene perception and signaling leads to upregulation of genes in anabolic pathways. We also associated complex phenotypes to subsets of differentially regulated genes.

Fermentation process for production of apple-based kefir vinegar: microbiological, chemical and sensory analysis

Abstract 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.

Phenolic Content and Antioxidant Activity during the Development of 'Brookfield' and 'Mishima' Apples.

The aim of this study was to characterize the changes in the contents of total (TPC) and individual (IPC) phenolic compounds, the total antioxidant activity (TAA) in the peel and pulp, and total anthocyanins (TAN) in the peel during the development of the fruits of 'Brookfield' and 'Mishima' apple trees. 'Brookfield' apples were harvested from the 49th to the 138th days after full bloom (DAFB) and 'Mishima' apples from the 45th to the 172th DAFB. In the pulp, the IPC, TPC, and TAA rapidly reduced at 75 and 79 DAFB for the 'Brookfield' and 'Mishima' apples, respectively, and then remained constant until commercial maturity. In the peel of 'Brookfield' apples there was a reduction in the TPC and TAA at 79 DAFB. The quercetin 3-galactoside, epicatechin, and procyanidin B2 contents reduced up to 107 DAFB with a subsequent increase in the values at commercial maturity. In the peel of 'Mishima' apples there was a reduction in the TPC, TAA, epicatechin, and procyanidin B1 and B2 contents at 130 DAFB, with a subsequent increase until commercial maturity. The TAN content in the peel increased during the 2 and 4 weeks prior to commercial maturity for 'Brookfield' and 'Mishima' apples, respectively. In the pulp and peel of both cultivars there was a reduction in the IPC, TPC, and TAA as the development proceeded. On nearing commercial maturity, there was an increase in the contents of quercetin 3-galactoside, epicatechin, procyanidin B2, and TAN in the peel for both cultivars.

A novel feedstuff: ensiling of cowpea (Vigna unguiculata L.) stover and apple (Malus domestica Borkh.) mixtures. Evaluation of the nutritive value, fermentation quality and aerobic stability.

BACKGROUND: Agro-industrial by-products are of low economic value as foods for human consumption but may have potential value as animal feedstuffs. This study evaluated a novel feedstuff, ensiled discarded apple (85%) and cowpea stover (15%) mixtures with two different ensiling periods (45 and 60 days), regarding the nutritive value, fermentation quality and aerobic stability. RESULTS: Generally, no differences (P > 0.05) were observed between ensiling periods for nutritive value and fermentation characteristics. Silages were stable after ensiling, presenting high lactic acid (77.3 g kg-1 dry matter (DM)) and acetic acid (54.7 g kg-1 DM) and low ethanol (15.7 g kg-1 DM) and NH3 -N (105.6 g kg-1 total N) concentrations. No butyric acid was detected in silages, and they were aerobically stable for up to 216 h. Lactic acid bacteria numbers were high at silo opening (7.14 log colony-forming units (CFU) g-1 ), while Enterobacteriaceae were not detected and yeasts/moulds were low (2.44 log CFU g-1 ). Yeast/mould and Enterobacteriaceae numbers grew considerably during 12 days of air exposure. CONCLUSION: A mixture of low calibre discarded apples with cowpea stover can be used as animal feed after the ensiling process owing to its nutritive value and long aerobic stability. © 2017 Society of Chemical Industry.

Fermentation process for production of apple-based kefir vinegar: microbiological, chemical and sensory analysis.

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 ∼41gL-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.

Physiological characterization of Saccharomyces uvarum and Saccharomyces eubayanus from Patagonia and their potential for cidermaking.

A diversity of yeast strains belonging to the cryotolerant fermentative species S. uvarum and S. eubayanus have been recovered from natural habitats and traditional fermentations in North Patagonia. The aim of this work was to evaluate the most relevant physiological features in a set of Patagonian strains belonging to S. uvarum and S. eubayanus, in order to analyze their potentiality to be used as starter cultures for cidermaking elaborated at low temperature. We evidenced that S. uvarum strains isolated from natural habitats (Araucaria araucana bark) showed similar physiological features to S. eubayanus strains obtained from the same habitat, and different from S. uvarum strains from fermentative environments (apple chichas). We also confirm the capacity of S. uvarum to produce high glycerol levels, low acetic acid and elevated production of the higher alcohol 2-phenylethanol and 2-phenylethyl acetate and demonstrated similar properties in S. eubayanus. Finally, we evidenced for the first time the antagonistic activity of S. eubayanus and selected three strains (two S. uvarum and one S. eubayanus) bearing the best combination of features to be used as a starter culture in cidermaking.