Optimization of a process for enzyme-assisted pigment extraction from grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.) pomace

A process for enzyme-assisted extraction of polyphenols from grape pomace was developed on laboratory and pilot-plant scale. After resuspending grape pomace in water, the skins were ground and the resulting mash was pasteurized to inactivate the deteriorative enzymes responsible for polyphenol degradation, and then pre-extracted with hot water. Subsequently, cell wall polysaccharides were hydrolyzed. The extract was separated from the solid residue by pressing, and finally spray dried. Before scaling-up, enzymatic hydrolysis was optimized on laboratory scale using a D-optimal design and analyzed by response surface methodology. A mixture of pectinolytic and cellulolytic enzyme preparations (ratio 2:1) yielded the highest amounts of phenolic compounds after 2 h of treatment, applying a dosage of 4,500 mg/kg (based on dry matter) at T = 40 °C and pH 4.0. Aqueous pre-extraction of the pomace followed by enzymatic treatment resulted in significantly improved extraction yields reaching 91.9, 92.4, and 63.6% for phenolic acids, non-anthocyanin flavonoids and anthocyanins, respectively. As the yields obtained were comparable to those from sulfite-assisted extraction, this process can be considered a suitable alternative to the application of sulfite.     

Factorial design optimisation of grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis>) seed polyphenol extraction

A 2³-full factorial design and response surface methodology were deployed to assess some basic factors (time, % ethanol and pH) affecting profoundly the extractability of polyphenolic phytochemicals from grape (V. vinifera) seeds. In an effort to obtain a thorough insight into the applicability of the models established, seed extracts from three different varieties were tested, by determining several indices of the polyphenolic composition, such as total polyphenol (TP), total flavanol (TFl) and proanthocyanidin (PC) concentrations. It has been shown that the models generated can adequately predict the recovery levels for each polyphenol group, but the optimal conditions predicted for TP, TFl and PC recovery varied significantly. Notable differences were also seen among the different varieties. Correlation of the polyphenol indices with the antiradical activity and reducing power of the extracts indicated that there is no consistent pattern to associate specific polyphenol classes with the antioxidant potential, which might be an evidence of antagonism. Examination of the optimally obtained extracts using liquid chromatography-mass spectrometry revealed that the most prominent compounds were catechin, some flavanol dimers and galloylated derivatives thereof.     

Quantification of DNA during winemaking by fluorimetry and <Emphasis Type="Italic">Vitis vinifera</Emphasis> L.-specific quantitative PCR

The ability to genotype wine would be of a great benefit to control the varietal wine authenticity. In this paper, we assessed the fate of grapevine DNA and total DNA during vinification to suggest the latest sampling point that would allow genotyping. The total DNA concentration was measured by fluorimetry and the specific Vitis vinifera L. DNA concentration was measured by quantitative PCR in samples obtained from different steps of vinifications of four wines. To elucidate the effect of pre-extraction and extraction steps on DNA yield from wine, several samples from bottle of wine, two precipitation salts, and two DNA extraction solutions were tested. It was found that hundreds of genome equivalents of grapevine DNA can be extracted from every winemaking sample and from wine. In tested samples, differences in DNA yield among wines from different grapevine varieties, differences among extraction methods, and sampling variation in wine were not statistically significant.     

Effect of glasswort (<Emphasis Type="Italic">Salicornia herbacea</Emphasis> L.) and rice (<Emphasis Type="Italic">oryza sativa</Emphasis> L.) on quality of <Emphasis Type="Italic">Ganjang</Emphasis> (Korean Soy Sauce)

Four types of meju were made from 100%(w/w) defatted soybean (DFS), a mixture of 80%(w/w) defatted soybean, and 20%(w/w) glasswort (DFS-G), a mixture of 80%(w/w) defatted soybean and 20%(w/w) rice (DFS-R), and a mixture of 60%(w/w) defatted soybean, 20%(w/w) glasswort, and 20%(w/w) rice (DFS-GR). Four types of Korean traditional soy sauce were prepared from the 4 types of meju. Mineral and antioxidant contents in the soy sauce made of DFS-G and DFS-GR were significantly higher than others. Citric, malic, succinic, lactic, and pyroglutamic acid contents in soy sauce made of DFS-R and DFS-GR were 1.3–1.5 times higher than others. Total nitrogen and free amino acid contents in soy sauce were correlated with DFS concentration in the meju. The bacterial community in the non-fermented meju-making ingredients was replaced largely by Bacillus sp. in the fermented meju. The use of glasswort and rice in the meju-making process did not alter the bacterial community responsible for the fermentation of meju.     

Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

Estrogenic effects of various extracts from <Emphasis Type="Italic">Chamdanggui</Emphasis> (<Emphasis Type="Italic">Angelica gigas</Emphasis> Nakai) and <Emphasis Type="Italic">sogdan</Emphasis> (<Emphasis Type="Italic">Phlomis umbrosa</Emphasis> Turcz)

The various extracts from chamdanggui (Angelica gigas Nakai) and sogdan (Phlomis umbrosa Turcz) were evaluated for estrogenic activity and characterized according to HPLC profile. Chamdanggui and sogdan were individually extracted with 4 solvents (hot water, 70% ethanol, n-butanol, and dichloromethane) of differing polarities. Estrogenic activity was determined by E-screen using an estrogen-dependent MCF-7 BUS cell. Although almost all extracts showed estrogenic effects in a concentrationdependent manner, the hot water extract from chamdanggui (250 μg/mL) had the higher effect (138%). Among 90 fractions using HPLC separation of the hot water extract from chamdanggui, fraction 21 and 28 produced the highest estrogenic effects of 178 and 163% at 10 μg/mL, respectively. The results imply that the hot water extract from chamdanggui could be useful as an alternative hormone replacement therapy.     

Characterisation of certain major polyphenolic antioxidants in grape (<Emphasis Type="Italic">Vitis vinifera</Emphasis> cv. Roditis) stems by liquid chromatography-mass spectrometry

The stem extract, which was shown to contain high amounts of phenolics and significant antioxidant potency, was further analysed employing liquid chromatography-electrospray ionisation mass spectrometry, in an effort to obtain a deeper insight into its polyphenolic composition. The compounds identified were mainly flavanols (a trimer and a galloylated dimer), flavonols (rutin and quercetin 3-O-glucuronide), stilbenes (trans-resveratrol and a resveratrol dehydrodimer) and a dihydroflavonol glycoside (astilbin).     

Effects of exogenous abscisic acid on phenolic characteristics of red <Emphasis Type="Italic">Vitis vinifera</Emphasis> grapes and wines

The effects of exogenous abscisic acid (ABA) on phenolic characteristics of grapes and wines were investigated in Vitis vinifera cvs. Merlot and Cabernet Sauvignon. Exogenous ABA treatment at veraison significantly improved phenolic contents, mainly anthocyanins and flavonols, and antioxidant properties of the grape skins, but had no effects on total phenolics and antioxidant activities in the seeds or on basic fruit qualities. The wines made from ABA-treated grapes were also consequently enhanced in total phenolics, anthocyanins, flavonols and antioxidant activities. The proportions of methylated anthocyanins in the skins and acylated anthocyanins and derived pigments in the wines were decreased to some degree by exogenous ABA treatment. No distinct relationships were observed between ABA concentrations and phenolic characteristics, and the effects were observed even with 200 mg/L ABA. The results revealed that exogenous ABA applied at veraison offered opportunities to improve phenolic contents and nutritional values of grape skins and wines.     

Enhanced selenium content in buckwheat (<Emphasis Type="Italic">Fagopyrum esculentum</Emphasis> Moench) and pumpkin (<Emphasis Type="Italic">Cucurbita pepo</Emphasis> L.) seeds by foliar fertilisation

The fruit and thin-husked seeds of the pumpkin (Cucurbita pepo L.) and buckwheat grain (Fagopyrum esculentum Moench), both grown in Slovenia, were analysed for selenium (Se) content following foliar application of Se(VI) solution during the period of blooming. Samples were digested by a H₂SO₄-HNO₃-H₂O₂-V₂O₅ mixture and Se determined, based on hydride generation atomic fluorescence spectrometry. The whole procedure from weighing to measuring was carried out in the same Teflon vessel. The detection limit of the method was 0.14 ng g^–1 solution. Buckwheat seeds from untreated plants contained 47 ng g^–1 of Se and 394 ng g^–1 from plants after foliar fertilisation with Se. Pumpkin seeds from untreated plants contained 108 ng g^–1 of Se, and 381 ng g^–1 of Se from Se-treated plants, all per lyophilised sample. Se content in lyophilised pumpkin fruit was 15 ng g^–1 in untreated plants and 20 ng g^–1 in Se-treated pumpkin plants. It is thus feasible to enhance Se content in buckwheat and pumpkin seeds by foliar fertilisation, making them a rich source of dietary Se and useful as a raw material for enriched food products.     

Effect of Vacuum Impregnation Combined with Calcium Lactate on the Firmness and Polysaccharide Morphology of Kyoho Grapes (<Emphasis Type="Italic">Vitis vinifera</Emphasis> x <Emphasis Type="Italic">V. labrusca</Emphasis>)

The effects of vacuum impregnation (VI) with 2% calcium lactate treatment on the VI properties (obtained from hydrodynamic mechanism and deformation–relaxation phenomena models), firmness, and pectin of Kyoho grapes were investigated. Fruit pectin was analysed by atomic force microscopy (AFM). VI was applied for 10–35 min at 25–45 °C and 5 kPa. The maximum values of effective porosity, ε_e (0.606%), and volume fraction, X (0.588%), occurred at 35 °C when the VI time was 15 min. No change was observed in the volumetric deformation (γ ≈ 0) of the grapes after the impregnation. The firmness significantly increased at 35 °C VI (from 12.93 to 14.47 N). According to the AFM results, calcium mainly inhibited the degradation of chelate-soluble pectin and sodium carbonate-soluble pectin short branches during the VI. Under the studied conditions, the validity of VI to incorporate calcium into fruit to improve the quality of grapes was verified, and a final corresponding product was obtained by VI.     

Optimization for encapsulation of green tea (<Emphasis Type="Italic">Camellia</Emphasis><Emphasis Type="Italic">sinensis</Emphasis> L.) extract by spray drying technology

Green tea has been credited with providing a wide variety of health benefits like higher total antioxidant, cancer prevention, and anti-irritant etc. Microencapsulation technology is used to prevent antioxidant loss during processing which involves entrapment of active material into carrier material. The experiments were designed using response surface methodology based on three level two factor (green tea extract, 10–25 % and drying inlet air temperature, 120, 130 and 140°C) central composite design. The feed emulsion was prepared with green tea extract as core material and 40 % maltodextrin concentration as wall material with the ratio of core to wall (1:2) and it was fed into the spray dryer at varied drying inlet air temperature to get encapsulated green tea extract powder. The quality characteristics were analysed such as moisture content, total antioxidant activity and total phenolic content and the process conditions were optimized using desirability function methodology. At optimum spray drying condition moisture content, total antioxidant activity and total phenolic content were found to be 3.1 % w.b., 72.91 % free radical scavenging activity by DPPH method, 57.81 mg/g of dry matter, expressed as gallic acid equivalent respectively with desirability value of 0.92.     

Study of Spray System Applications of Edible Coating Suspensions Based on Hydrocolloids Containing Cellulose Nanofibers on Grape Surface (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.)

An edible coating is a useful technology to preserve fruit quality by covering them with a protective layer which improves the appearance and provides a semipermeable barrier for gases and water vapor transfer, allowing extension of their shelf life. In this study, edible coatings based on hydroxypropyl methyl cellulose, k-carrageenan, glycerol, and cellulose nanofibers were formulated. The operational conditions of the spray system were studied to obtain a coating with optimal adhesion on grape (Vitis vinifera L.) surface. Furthermore, the physico-chemical properties of grapes covered with edible coatings were evaluated during refrigerated storage. A full factorial 2³ experimental design was applied, where liquid suspension flow (1–5 L h^?1), air pressure (50–200 kPa), and height of impact (0.3–0.5 m) were evaluated as independent variables, whereas the percentage of coating and thickness of coating was the response variables. Both mechanical and physico-chemical properties were evaluated during 41 days in both coated and uncoated grapes as shelf life criteria. Throughout the storage time, noticeable changes in pH and total soluble solids were not found in grapes covered with edible coatings and they showed the highest stability for the evaluated mechanical properties. Moreover, coated grapes showed final weight loss and water vapor permeability values of approximately 30 and 34% lower, respectively, than uncoated grapes, suggesting a shelf life extension.     

Effects of suni-bug (<Emphasis Type="Italic">Eurygaster</Emphasis> spp.) damage on size distribution of durum wheat (<Emphasis Type="Italic">Triticum durum</Emphasis> L.) proteins

Wholemeal samples were obtained from five durum wheat cultivars at two different bug (Eurygaster spp.) damage levels (medium and high damage). The samples were incubated (60 and 120 min) and used in size exclusion high performance liquid chromatography (SE-HPLC) analyses. The results showed that the amount of larger polymeric protein (TP1) and smaller polymeric protein (TP2) obtained from total (sodium dodecyl sulfate soluble) proteins decreased significantly in the bug-damaged samples, while the amount of total larger monomeric proteins (TP3) increased. The polymeric/monomeric protein ratio of all cultivars decreased at 60 min of incubation with increasing damage level. For all cultivars, the ratio of unextractable polymeric protein (UPP%) significantly decreased at 60 min of incubation except cv. Diyarbakir. The results suggested that bug protease caused depolymerization and/or disaggregation of polymeric proteins to lower their average molecular size. The changes in protein structure as determined using SE-HPLC supported by the decreases in gluten content and gluten index values which decreased with suni-bug damage. Deteriorative effects of bug damage on durum wheat quality were found to be quite similar to those on bread wheats.     

Aromatic characterization and enological potential of 21 minor varieties (<Emphasis Type=Italic>Vitis vinifera</Emphasis> L.)

The homogenization of international wine market led to a gradual impoverishment of the genetic pool. In fact, in several Spanish Quality Demarcations, the most frequent international varieties are replacing the local ones. As a result, minor varieties, perfectly adapted to the local environmental conditions, are nowadays at risk of extinction. The study of minor varieties could provide useful inputs to satisfy the demand for new and interesting wine products. This work aims at filling a gap in the existing literature, focusing on the aromatic potential of minor varieties. Here, the study of glycosidic volatile compounds and the evaluation of the influence of several variables on aroma composition were considered. Fifty-one glycosidic compounds were identified and quantified. The results revealed differences for glycosidic compounds according to cultivars, berry color, clone and sample origin. The inclusion or not of the cultivars in Quality Demarcation was also discussed. Moreover, the synthesis of some compounds involved in these differentiations seems to have a genetic component. The characterization of aromatic potential of several minor varieties, achieved for the first time, revealed that some of these (Argamusa, Quigat, Pampolat girat, Excursach, Gorgollasssa or Mandón) could represent an excellent option for winemaking and commercial offer diversification strategies, besides being important for these cultivars conservation.     

Thermal inactivation kinetics of quality-related enzymes in cauliflower (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">botrytis</Emphasis><Emphasis Type="Bold">)</Emphasis>

Thermal inactivation of quality-related enzymes in both cauliflower crude enzyme extracts and fresh tissue samples was studied in temperature range 50–100 °C. For crude enzyme extracts, several parameters, reaction rate constants (k) and activation energy (E _a) as well as decimal reduction time (D) and (z) values, were used to characterize the thermal stability. The rates of inactivation were found to follow first-order inactivation kinetics. Activation energies varied between 101.18 and 208.42 kJ mol⁻¹ with z values of 10.59–24.09 °C. The examined kinetics indicated that lipoxygenase was the most heat resistant followed by peroxidase, polyphenol oxidase, pectin methyl esterase and ascorbic acid oxidase. Furthermore, the obtained results from the blanched fresh tissues indicated that inactivation of lipoxygenase secured disappearing of any other enzyme activities. Therefore, this study recommends using lipoxygenase as an indicator enzyme to optimize the thermal treatments of cauliflower products.     

A New Multiplexed Real-Time PCR Assay to Detect <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>, <Emphasis Type="Italic">C. coli</Emphasis>, <Emphasis Type="Italic">C. lari</Emphasis>, and <Emphasis Type="Italic">C. upsaliensis</Emphasis>

A new rapid method based on real-time PCR was developed to detect four thermophilic Campylobacter species (Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis) in food samples. The assay targeted the bipA gene for C. upsaliensis and C. lari, whereas the gene encoding the ATP-binding protein CJE0832 was used to detect C. coli and C. jejuni. These genes were chosen for this assay due to their low variability and mutation rate at a species level. The multiplex PCR showed 100% inclusivity for all 25 thermophilic Campylobacter strains tested and 100% exclusivity for 38 non-targeted strains belonging to closely related species. The newly developed real-time PCR could detect down to 10² genomes/reaction and displayed efficiency above 97% for all species except for C. upsaliensis (90.1%). The method proved to be a reliable tool for food analysis, showing 100% sensitivity, 96% efficiency, and 92.45% specificity when validated against the gold standard method UNE-EN ISO 10272:2006 using 200 diverse food samples (meat, fish, fruits and vegetables, and raw milk). In artificially spiked samples, the detection limit of the method was 10 cfu/g in salad, 5 cfu/g in turkey meat, and 1 cfu/g in the rest of meat samples tested. Consequently, the newly designed molecular tool represents a quick and safe alternative to obtain reliable results concerning the presence/absence of the main thermophilic Campylobacter in any food sample.     

In vivo study of antiallergenicity of ethanol extracts from <Emphasis Type="Italic">Sargassum tenerrimum</Emphasis>, <Emphasis Type="Italic">Sargassum cervicorne</Emphasis> and <Emphasis Type="Italic">Sargassum graminifolium turn</Emphasis>

Food allergy has becoming the serious threat in the world for which the search of an effective anti-allergic drug is the demand of time. Keeping in view of the potentiality of seaweeds, the ethanol extracts from Sargassum tenerrimum (ST), Sargassum cervicorne (SC), and Sargassum graminifolium turn (SG) have been studied in vivo for its antiallergenicity through passive cutaneous anaphylaxis (PCA) and active cutaneous anaphylaxis (ACA) in female BALB/c mice. Intraperitoneal administration of these ethanol extracts inhibit mouse PCA and ACA in a dose-dependent manner using ovalbumin (OVA) and shrimp allergen as triggering agents to induce allergenicity over mice. The extract of ST containing phlorotannin has been found most active over the suppression of PCA triggered by OVA and shrimp with IC₅₀ values of 25.64 and 40.98 mg/kg, respectively and an efficacy comparable to that of an anti-allergic drug disodiumcromoglycate. Similarly, ST inhibits ACA triggered by ova and shrimp allergen in the mouse, with 50% suppression at 25.5 and 43.53 mg/kg, respectively. The results presented here show that these extracts are active on the studied models among which ethanol extract of ST was the most potent, leading toward the promising development of a new class of anti-allergic drugs.     

Molecular diversity among natural populations of <Emphasis Type="Italic">Lactobacillus paracasei</Emphasis> and <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis>/<Emphasis Type="Italic">paraplantarum</Emphasis> strains isolated from autochthonous dairy products

The diversity of 87 Lactobacillus paracasei and Lactobacillus plantarum/paraplantarum strains, previously identified from different autochthonous dairy products, was investigated by phenotypic and genotypic approaches. The increased resolution obtained using phenotypic and genotypic characterization allowed the level of strain heterogeneity detection to be widened. Phenotypic diversity was evaluated by studying biochemical characteristics of technological interest, including antimicrobial and proteinase activities, resistance to nisin, aggregation ability, production of exopolysaccharides, acetoin and diacetyl, citrate utilization, and antibiotic susceptibility. Genotypic diversity was generally evaluated by PCR amplification of repetitive bacterial DNA element fingerprinting using the (GTG)₅ primer [(GTG)₅-PCR]. Moreover, in cases where strains were not discriminated by (GTG)₅-PCR combined with phenotypic analysis, pulsed-field gel electrophoresis (PFGE) analysis was performed. The results indicate that L. plantarum/paraplantarum and L. paracasei natural isolates from artisanal dairy products are a gold mine in terms of diversity of strains and could be potentially interesting to dairy companies for the formulation of functional starter cultures in the production of innovative foods.     

Antioxidant and emulsion properties of freshwater carps (<Emphasis Type="Italic">Catla catla</Emphasis>, <Emphasis Type="Italic">Labeo rohita</Emphasis>, <Emphasis Type="Italic">Cirrhinus mrigala</Emphasis>) protein hydrolysates prepared using flavorzyme

Fish protein hydrolysates (FPHs) were prepared from freshwater carps (Catla catla, Labeo rohita, and Cirrhinus mrigala) using flavorzyme at different degrees of hydrolysis (DHs) ranging from 5 to 20%. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free-radical-scavenging activity of the FPHs prepared from the three species were in the range of 50–82%; the ferric reducing power of the FPHs prepared from catla was the highest. The linoleic acid peroxidation inhibition activity of the prepared FPHs varied from 71 to 91%. The emulsion activity index of the FPHs prepared from catla and rohu decreased significantly with an increase in the DH (p < 0.05). The emulsion stability index of the FPHs prepared from the three species was the highest at 20% DH. FPHs prepared from freshwater carps possess good antioxidant and surface-active properties and are therefore suitable to be used as natural antioxidants in health-food formulation and as water-soluble antioxidants in the food-processing industry.