Biotransformation of hop derived compounds by Brettanomyces yeast strains

Several hop derived compounds in wort are known to be converted by yeast during fermentation, influencing the overall perception of the beer. A deeper understanding of such metabolic processes during fermentation is needed to achieve better control of the outcome. Here, the interaction between hop derived compounds and the yeast genera Brettanomyces was studied. Several Brettanomyces strains with different genomic backgrounds were selected, focusing on two traits: beta-glucosidase activity and nitrate assimilation. The role of three beta-glucosidases present in Brettanomyces bruxellensis and Brettanomyces anomalus and their impact on the final monoterpene alcohol profile was analysed. The beta-glucosidase activity was highly strain dependent, with B. anomalus CRL-49 exhibiting the highest conversion. Such activity could not be related to the release of aglycones from hops during fermentation, as a substantial part of such activity was intracellular. Nevertheless, the reduction of geraniol to β-citronellol was remarkably efficient for all Brettanomyces strains during fermentation, and it is suggested that two oxidoreductases BbHye2 and BbHye3 may have an influence. Moreover, the transfer of nitrate from hops to wort and its further assimilation by Brettanomyces species was analysed. The amount of nitrate in wort proved to be linearly proportional to the contact time of the hops with the wort. The level of nitrate assimilation by yeast was shown to be dependent on the nitrate assimilation cluster (YNR, YNI, YNT). Hence, the desired yeast strains may be selected according to the genetic make-up. © 2020 The Authors. Journal of the Institute of Brewing published by John Wiley & Sons Ltd on behalf of The Institute of Brewing & Distilling     

5‐(Piperidin‐4‐yl)‐3‐Hydroxypyrazole: A Novel Scaffold for Probing the Orthosteric γ‐Aminobutyric Acid Type A Receptor Binding Site

A series of bioisosteric N₁‐ and N₂‐substituted 5‐(piperidin‐4‐yl)‐3‐hydroxypyrazole analogues of the partial GABA_AR agonists 4‐PIOL and 4‐PHP have been designed, synthesized, and characterized pharmacologically. The unsubstituted 3‐hydroxypyrazole analogue of 4‐PIOL ( 2 a ; IC₅₀～300 μM ) is a weak antagonist at the α₁β₂γ₂ GABA_AR, whereas substituting the N₁‐ or N₂‐position with alkyl or aryl substituents resulted in antagonists with binding affinities in the high nanomolar to low micromolar range at native rat GABA_ARs. Docking studies using a α₁β₂γ₂ GABA_AR homology model along with the obtained SAR indicate that the N₁‐substituted analogues of 4‐PIOL and 4‐PHP, 2 a – k , and previously reported 3‐substituted 4‐PHP analogues share a common binding mode to the orthosteric binding site in the receptor. Interestingly, the core scaffold of the N₂‐substituted analogues of 4‐PIOL and 4‐PHP, 3 b – k , are suggested to flip 180° thereby adapting to the binding pocket and addressing a cavity situated above the core scaffold.     

A deep learning approach for the forensic evaluation of sexual assault

Despite the existence of patterns able to discriminate between consensual and non-consensual intercourse, the relevance of genital lesions in the corroboration of a legal rape complaint is currently under debate in many countries. The testimony of the physicians when assessing these lesions has been questioned in court due to several factors (e.g., a lack of comprehensive knowledge of lesions, wide spectrum of background area, among others). Therefore, it is relevant to provide automated tools to support the decision process in an objective manner. In this work, we evaluate the performance of state-of-the-art deep learning architectures for the forensic assessment of sexual assault. We propose a deep architecture and learning strategy to tackle the class imbalance on deep learning using ranking. The proposed methodologies achieved the best results when compared with handcrafted feature engineering and with other deep architectures.     

Quality of Sour Cream Packaged in Cups with Different Light Barrier Properties Measured by Fluorescence Spectroscopy and Sensory Analysis

ABSTRACT:  In this study, photooxidation of sour cream packaged in cups with different light barrier properties was investigated. The sour cream was light exposed for 36 h with a standard fluorescent light tube, simulating storage conditions in many Norwegian grocery stores. Three different cups were evaluated: a white cup, a cup with medium light barrier (LB), and a cup with high LB. The quality of the sour cream was evaluated by sensory analysis and front face fluorescence spectroscopy. The sour cream stored in white cups became very rancid during the light exposure, with a rancid flavor score of 7.8 on a scale from 1 to 9, where 9 is the highest intensity. Cups with high LB gave best protection against the light. The sensory assessors could not distinguish between sour cream stored in cups with high LB and sour cream stored in the dark. The rancid flavor intensity for the cups with medium LB was significantly different ( P  < 0.05) from both the white cup and the cup with high LB. The sensory flavor intensities indicated that with 36 h of light exposure, only a cup with high LB will protect the sour cream sufficiently. The fluorescence measurements corresponded well with the sensory analysis as the photosensitizers were somewhat degraded for sour cream stored in cups with medium LB and most degraded in the white cups. For the cups with high LB, the fluorescence peaks that originated from the photosensitizers were all intact.
PRACTICAL APPLICATION: Dairy products such as sour cream will develop off-odor and taste faults ("sunlight taste") in a few hours if exposed to sufficient fluorescent lightning in the grocery stores. A light barrier can be incorporated in the packaging material to protect the dairy products from the light. Our experiment showed that the incorporation of a black pigment into 1 of the 3 plastic layers in the cup for packaging of sour cream protected the sour cream from degradation when exposed to light.     

Pre-treatment and ethanol fermentation potential of olive pulp at different dry matter concentrations

Renewable energy sources have received increased interest from the international community with biomass being one of the oldest and the most promising ones. In the concept of exploitation of agro-industrial residues, the present study investigates the pre-treatment and ethanol fermentation potential of the olive pulp, which is the semi solid residue generated from the two-phase processing of the olives for olive oil production. Wet oxidation and enzymatic hydrolysis have been applied aiming at the enhancement of carbohydrates' bioavailability. Different concentrations of enzymes and enzymatic durations have been tested. Both wet oxidation and enzymic treatment were evaluated based on the ethanol obtained in a subsequent fermentation step by Saccharomyces cerevisiae and Thermoanaerobacter mathranii. It was found that a four-day hydrolysis time was adequate for a satisfactory release of glucose and xylose. The combination of wet oxidation and enzymatic hydrolysis resulted in the glucose and xylose concentration increase of 138 and 444%, respectively, compared to 33 and 15% with only enzymes added. However, the highest ethanol production was obtained when only enzymic pre-treatment was applied, implying that wet oxidation is not a recommended pre-treatment process for olive pulp at the conditions tested. It was also showed that increased dry matter concentration did not have a negative effect on the release of sugars, indicating that the cellulose and xylan content of the olive pulp is relatively easily available. The results of the experiments in batch processes clearly emphasize that the simultaneous saccharification and fermentation (SSF) mode is advantageous in comparison with the separate hydrolysis and fermentation (SHF) mode concerning process contamination.     

Development and application of new nucleic acid-based technologies for microbial community analyses in foods

Several challenges still persist in the analysis of microorganisms in foods, particularly in studies of complex communities. Nucleic acid-based methods are promising tools in addressing new questions concerning microbial communities. We have developed several new methods in the field of nucleic acid-based microbial community analyses. These methods cover both sample preparation and detection approaches. The sample preparation method involves simplified DNA purification using paramagnetic beads. As an extension of this method, the same paramagnetic beads are used for both cell separation and DNA purification. This enables full automation. The separate detection of viable and dead bacteria is a major issue in nucleic acid-based diagnostics. We have applied a living/dead dye that binds covalently to DNA and inhibits the PCR from dead cells. In addition, a DNA array-based detection assay has been developed. The assay combines the specificity obtained by enzymatic labeling of DNA probes with the possibility of detecting several targets simultaneously by DNA array hybridization. In combination with 16S rDNA amplification, this is a promising tool for community analyses. Also, we have developed a novel approach for multiplex quantitative PCR. The multiplex PCR has been combined with our DNA array-based detection method. Finally, we are now in the process of adapting a system for monitoring microbial growth and death in real-time through the tagging of bacteria with green fluorescent protein (GFP) combined with fluorescence detection using a high-resolution confocal laser scanner.     

The effect of reuterin on the lag time of single cells of Listeria innocua grown on a solid agar surface at different pH and NaCl concentrations

The lag time of single cells of Listeria innocua grown on the surface of Brain Heart Infusion Agar was studied by microscopy and image analysis. An experimental set-up that enabled relocation of the cells on the agar surface was developed and used to collect data from 50 to 100 individual cells at a time. Reuterin was added at different concentrations (0-10 AU/ml) and it was observed that it increased both the lag time of the cells and its variance. Furthermore, for a large proportion of cells, reuterin completely prevented the cell division within the time of observation. Reuterin in combination with low pH inhibited the cell division even more efficiently. A similar effect was observed for the combination of reuterin and sodium chloride. Our experimental set-up provides a good model system for generating data on the lag time of single cells on solid surfaces, which can improve the predictions of microbial growth on solid food matrices.     

Real-time PCR quantification of the AM-toxin gene and HPLC qualification of toxigenic metabolites from Alternaria species from apples

Some Alternaria species are able to produce plant pathogenic as well as toxic metabolites. In both agriculture and the food industry it is important know if toxigenic Alternaria are present to rapidly employ the correct corrective actions. The purpose of this work was to establish a real-time PCR method, which can detect and quantify apple pathogenic and toxigenic Alternaria. An AM-toxin I primer set, which could recognize Alternaria DNA only, was designed by using primers complementary to the AM-toxin I gene. The method could detect small amounts of DNA (4 pg) and still obtain a large dynamic range (4 decades) without interference from apple material. Eight Alternaria isolates were analyzed for the presence of AM-toxin I gene and their production of secondary metabolites. Then analyses showed that all eight isolates contained the AM toxin gene and were able to produce the plant pathogenic tentoxin in addition to AM toxin I. The analyses also showed the production of tenuazonic acid, alternariols, Altenuene, altenusin and/or altertoxin I in pure culture. Analyses of inoculated apples showed that both the AM-toxin gene and alternariol monomethyl ether could be detected. Morphological analyses suggested that the eight Alternaria strains, though they all carried the AM toxin genes, probably belong to different but closely related un-described Alternaria taxa in the A. tenuissima species-group based on morphological and chemical differences.     

Wet oxidation pre‐treatment of woody yard waste: parameter optimization and enzymatic digestibility for ethanol production

Woody yard waste with high lignin content (22% of dry matter (DM)) was subjected to wet oxidation pre‐treatment for subsequent enzymatic conversion and fermentation. The effects of temperature (185–200 °C), oxygen pressure (3–12 bar) and addition of sodium carbonate (0–3.3 g per 100 g DM biomass) on enzymatic cellulose and hemicellulose (xylan) convertibility were studied. The enzymatic cellulose conversion was highest after wet oxidation for 15 min at 185 °C with addition of 12 bars of oxygen and 3.3 g Na₂CO₃ per 100 g waste. At 25 FPU (filter paper unit) cellulase g^?1 DM added, 58–67% and 80–83% of the cellulose and hemicellulose contained in the waste were converted into monomeric sugars. The cellulose conversion efficiency during a simultaneous saccharification and fermentation (SSF) assay at 10% DM was 79% for the highest enzyme loading (25 FPU g^?1 DM) while 69% conversion efficiency was still reached at 15 FPU g^?1 DM. Total carbohydrate recoveries were high (91–100% for cellulose and 72–100% for hemicellulose) and up to 49% of the original lignin and 79% of the hemicellulose could be solubilized during wet oxidation treatment and converted into carboxylic acids mainly (total carboxylic acids = 3.1–7.4% on DM basis). Copyright © 2004 Society of Chemical Industry