Genetic analysis of geraniol metabolism during fermentation

Geraniol produced by grape is the main precursor of terpenols which play a key role in the floral aroma of white wines. We investigated the fate of geraniol during wine fermentation by Saccharomyces cerevisiae. The volatile compounds produced during fermentation of a medium enriched with geraniol were extracted by Stir-bar sorptive extraction and analysed by GC–MS. We were able to detect and quantify geranyl acetate but also citronellyl- and neryl-acetate. The presence of these compounds partly explains the disparition of geraniol. The amounts of terpenyl esters are strain dependant. We demonstrated both by gene overexpression and gene-deletion the involvement of ATF1 enzyme but not ATF2 in the acetylation of terpenols. The affinity of ATF1 enzyme for several terpenols and for isoamyl alcohol was compared. We also demonstrated that OYE2 is the enzyme involved in geraniol to citronellol reduction. Fermenting strain deleted from OYE2 gene produces far less citronellol than wild type strain. Moreover lab strain over-expressing OYE2 allows 87% geraniol to citronellol reduction in bioconversion experiment compared to about 50% conversion with control strain.     

Antioxidant properties of durian fruit as influenced by ripening

The antioxidant properties of durian (Durio zibethinus Murr., cv. Mon Thong) at different stages of ripening were investigated using fluorometry, UV spectroscopy, and HPLC/DAD analyses. Total polyphenols, flavonoids, anthocyanins and flavanols in ripe durian were significantly higher (p < 0.05) than in mature and overripe fruits. Free polyphenols and flavonoids were at lower levels than hydrolyzed ones. Caffeic acid and quercetin were the dominant antioxidant substances in ripe durian. In these fruits, methanol extracts contained a relatively high capacity of 74.9 ± 7.1% inhibition using β-carotene-linoleic acid assay. Ferric-reducing/antioxidant power (FRAP) and cupric-reducing antioxidant capacity (CUPRAC) assays supported this finding. The correlation coefficients between polyphenols and antioxidant capacities of durian samples with all applied assays were about 0.98. In conclusion, the bioactivity of ripe durian was high and the total polyphenols were the main contributors to the overall antioxidant capacity.     

Survival of Listeria monocytogenes in a simulated dynamic gastrointestinal model during storage of inoculated bologna and salami slices in vacuum packages

Listeria monocytogenes counts were determined during storage (82 days, 4 degrees C) in vacuum packages of inoculated bologna and salami slices and after exposure to a simulated dynamic model of the stomach and small intestine. Variables controlled in the model included gastric emptying and gastrointestinal fluid secretion rates, gradual gastric acidification, and intestinal pH maintenance. L. monocytogenes populations increased on bologna and decreased on salami, reaching 8.7 and 1.4 log CFU/g, respectively, on day 82. Inactivation rates (IR) during gastric exposure of bologna and salami ranged from 0.079 (day 14) to 0.158 (day 57) log CFU/g/min and from 0.013 (day 42) to 0.051 (day 1) log CFU/g/min, respectively. On corresponding days, gastric IR for cells on salami were lower than on bologna, suggesting potential protective effects of the former product. However, it is also possible that the low initial L. monocytogenes levels reached with storage of salami (< or = 2.5 log CFU/g after day 27) may have resulted in slower reductions than in the high levels on bologna. Gradual decline of gastric pH allowed survival in the gastric compartment during the initial stages, which resulted in a large fraction of the cells being delivered into the intestinal compartment. Intestinal IR ranged from 0.003 to 0.048 (bologna) and from 0.002 to 0.056 (salami) log CFU/g/min throughout storage. Although findings indicated potential effects of salami against gastric killing of L. monocytogenes, any effects of the food matrix per se on the gastrointestinal survival of the pathogen were overwhelmed by the high and low contamination levels reached on bologna and salami, respectively, during storage.     

Free amino acids and biogenic amines in Alicante Monastrell wines

The simultaneous determination of 17 free amino acids and 8 biogenic amines in Alicante Monastrell wines was investigated for the first time. The quantification was carried out by using a RP-HPLC method, based on a pre-column derivatization with o-phthaldialdehyde (OPA) and fluorescence detection. From the results obtained it may be concluded that the most abundant free amino acids were Glu, Arg, Ala Asp, and Lys. None of the wine samples analysed had histamine (HIM) or Tyramine (TYM) levels above the limits considered as a possible toxic risk for healthy individuals. No measurable amounts of cadaverine (CAD) or methylamine (MEA) were found, showing no spoilage symptoms of sensory properties of the wines. Tryptamine (TRM) content was significantly higher in aged wines compared to young wines. However ethanolamine (ETA) content was lower. These data were used to make a preliminary classification of the samples using cluster analysis.     

Salmonella and Campylobacter biofilm formation: a comparative assessment from farm to fork

It takes several steps to bring food from the farm to the fork (dining table), and contamination with food‐borne pathogens can occur at any point in the process. Campylobacter spp. and Salmonella spp. are the main microorganisms responsible for foodborne disease in the EU. These two pathogens are able to persist throughout the food supply chain thanks to their ability to form biofilms. Owing to the high prevalence of Salmonella and especially of Campylobacter in the food supply chain and the huge efforts of food authorities to reduce these levels, it is of great importance to fully understand their mechanisms of persistence. Diverse studies have evaluated the biofilm‐forming capacity of foodborne pathogens isolated at different steps of food production. Nonetheless, the principal obstacle of these studies is to reproduce the real conditions that microorganisms encounter in the food supply chain. While there are a wide number of Salmonella biofilm studies, information on Campylobacter biofilms is still limited. A comparison between the two microorganisms could help to develop new research in the field of Campylobacter biofilms. Therefore, this review evaluates relevant work in the field of Salmonella and Campylobacter biofilms and the applicability of the data obtained from these studies to real working conditions. © 2018 Society of Chemical Industry     

Microwave and pulsed electric field assisted extractions of polyphenols from defatted canola seed cake

The incorporation of microwave and pulsed electric field (PEF) technologies in optimising the extraction of polyphenols from defatted canola seed cake was determined by Box‐Behnken response surface methodology, using three parameters of microwave treatment (power, time and liquid: solid (L:S) ratio) and four parameters of PEF (ethanol concentration, time, frequency and voltage). After microwave and PEF pretreatments, the polyphenols of defatted canola seed cake were extracted by ultrasound treatment under fixed variables (200 W of ultrasonic power, 70 °C of water bath and 20 min of extraction time). The measured responses were total phenolics, total flavonoids, the DPPH˙ scavenging activity and ferric reducing antioxidant power (FRAP). The maximum yields of all responses were achieved under optimum conditions of microwave processing (5 min, L:S ratio of 6.0 and 633.3 W) and PEF‐assisted extraction (30 V, 30 Hz, 10% ethanol concentration and 10 s).     

Study of the inhibitory effect of hydrophobic fluorescent markers on the enzymic coagulation of bovine casein micelles: action of TNS

The interaction of 2-(p-toluidinyl) naphthalene-6-sulfonate (TNS) with casein micelles (CM) was studied by fluorescence spectroscopy. Fluorescence emission spectra of the complex showed blue shift and intensity enhancement of TNS fluorescence, suggesting the insertion of the marker in low polarity regions of CM. An energy transfer process between the proteins and the marker was detected, showing that most of the TNS binding sites were in the proximity of CM fluorescent residues. TNS inhibited the aggregation step of CM enzymic coagulation, producing probably a decrease of size and amount of aggregates formed. This effect could not be related only to changes in CM net charge, but also possibly to the occupancy of surface hydrophobic regions by the marker. About a 20% decrease in the TNS fluorescence intensity was observed during the proteolytic step of coagulation which could be attributed to release of the marker from its binding sites located in the CM external layer. A bound TNS release was also observed during the initial time of the aggregation step, probably by removal of the bound marker from contact regions between aggregating particles upon their collisions. A further increase, related to the aggregation step, could indicate the uptake of the marker by new hydrophobic sites created in the complex structure of the clusters. The results pointed to the participation of surface hydrophobic regions of renneted CM in their aggregation process.     

Spectroscopic study of copper(II)--wheat straw cell wall residue surface complexes

The sorption of copper(II) by wheat straw cell wall residue (CWR) was studied and revealed a relatively stable surface complexation on the acid sites of the substrate (carboxylic and phenolic moieties). The copper binding capacity at pH = 5.75 and ionic strength of 0.1 M was evaluated at 63 micromol x g(-1) CWR. The Langmuir and Freundlich isotherm equations were then used to describe the partitioning behavior for the system at different pH values. Batch experiments have been achieved in the presence of calcium in order to mimic the calcareous soil of the Champagne region. A competitive effect on copper complexation has been shown, which is presumably due to the calcium ability to form outer-sphere complexes far less stable than copper(II) ones. Electron spin resonance and X-ray absorption spectroscopy were combined to obtain information on the geometry and structure of Cu bound to CWR. At least two different binding sites for Cu2+ were found to take place in CWR. From ESR parameters, we deduced that copper(II) ions, when complexed with CWR, are coordinated in a square-planar arrangement with four oxygen-containing groups. EXAFS and XANES experiments revealed that Cu(II) is surrounded by four oxygen atoms, with an average Cu-O equatorial distance equal to 1.94 A.