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.     

Pressurized acidified water extraction of black carrot [<Emphasis Type="Italic">Daucus carota</Emphasis> ssp. <Emphasis Type="Italic">sativus</Emphasis> var. <Emphasis Type="Italic">atrorubens</Emphasis> Alef.] anthocyanins

The anthocyanins present in black carrot were extracted with pressurized water acidified with sulfuric, citric and lactic acids. Anthocyanin degradation became significant above 100 °C and there was no improvement when extraction pressure was increased to 100 bar. Therefore, the extraction from black carrot was carried out at temperatures 50, 75 and 100 °C under 50 bar pressure. The extraction efficiencies in terms of acylated and non-acylated anthocyanins were comparable for all three acids used to acidify water at 50 °C, while similar results were observed at 75 °C for both citric and lactic acids. Water acidified with lactic acid showed significantly higher extraction efficiency at 100 °C compared to water acidified with sulfuric and citric acids. Highest degree of polymerization together with increasing degree of browning was observed within extracts when sulfuric acid was used. On the other hand, when organic acids were used to acidify water, a higher extraction efficiency of anthocyanins, accompanied with a relatively low polymerization and browning was observed, with lactic acid giving the best results.     

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.     

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.     

Differences of Bactericidal Efficacy on <Emphasis Type="Italic">Escherichia coli</Emphasis>, <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>, and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> of Slightly and Strongly Acidic Electrolyzed Water

In the present study, the disinfection efficacy of slightly acidic electrolyzed water (SAEW) and strongly acidic electrolyzed water (AEW) was tested on three bacteria, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis and the disinfection mechanism was discussed. The results showed that SAEW had a stronger antibacterial efficacy against these tested bacteria in comparison with AEW. The results also showed that both SAEW and AEW treatments could damage the cell membrane, which was demonstrated microcosmically by scanning electron microscopy (SEM), thus causing leakages of protein, DNA, RNA, and ATP, resulting in the death of microbes. Moreover, AEW treatment could not cause the degradations of DNA and RNA, and nucleic acids including DNA and RNA are not the target point of its bactericidal efficacy. However, SAEW could maybe cause the degradation of RNA, and RNA may be the target in its antibacterial activity. We suggested that the differences in antibacterial efficacy between SAEW and AEW could be explained by the different impacts on RNA of tested strains.     

Xanthan gum production under different operational conditions by <Emphasis Type="Italic">Xanthomonas axonopodis</Emphasis> pv <Emphasis Type="Italic">vesicatoria</Emphasis> isolated from pepper plant

The influence of operational conditions (pH, stirrer speed, and temperature) used in the process of xanthan production by Xanthomonas axonopodis pv vesicatoria (XCVA3-1) isolated from pepper plant were evaluated through yield of xanthan and compared with control strain Xanthomonas campestris NRRL-B 1459. Different conditions used during the fermentation affected the xanthan production. In this study the best combination of yield was obtained, reaching 1.325 g/100 mL with the use of pH 7.0, 30°C, and 250 rpm during fermentation. Increased yield of xanthan production can be obtained at high agitation values, with the maximum at 400 rpm. Higher yields of gum production can be obtained at 30°C and the optimum pH was found 7.0. This results were similar for the X. campestris NRRL-B 1459.     

Spray Chilling Microencapsulation of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> and <Emphasis Type="Italic">Bifidobacterium animalis</Emphasis> subsp. <Emphasis Type="Italic">lactis</Emphasis> and Its Use in the Preparation of Savory Probiotic Cereal Bars

The use of probiotic microorganisms has been limited by the difficulty of maintaining their viability during processing and throughout the product’s shelf life. This study evaluated the viability of microencapsulating Lactobacillus acidophilus (LA) and Bifidobacterium animalis subsp. lactis (BL) using the spray chilling technique to add them to savory cereal bars. The results showed that spray chilling generated a powder that was composed of smooth and continuous spheres with low moisture content and low water activity. The microencapsulated microorganisms exhibited a storage viability at least of 90 days as microparticles and in savory cereal bars, and their counts were superior to those resulting from other methods of adding activated and lyophilized probiotics to savory cereal bars. Thus, microparticles prepared by spray chilling are good vehicles for incorporating probiotics into cereal bars and have the potential to release the probiotics in the consumers’ intestines by means of fat digestion. Savory cereal bars that did and did not contain probiotics exhibited no differences in sensorial acceptance or commercial potential.     

Drying kinetics of whole and sliced <Emphasis Type="Italic">shiitake</Emphasis> mushrooms (<Emphasis Type="Italic">Lentinus edodes</Emphasis>)

Isotherms of shiitake mushroom (Lentinus edodes) at 25 and 40°C were determined and drying kinetics of whole and sliced shiitake mushrooms were tested using a convective air drying method at different drying temperature of 40, 50, 60, and 70°C. The monolayer moisture contents of the mushroom were 7.23 and 5.44 g water/100 g of dry solids at 25 and 40°C, respectively. Both mushroom samples showed falling drying rate periods with increasing drying rates with increases in drying temperature, and the drying rate of the sliced mushrooms was faster than that of the whole mushrooms at the same drying conditions. The kinetic parameters for dehydration of the mushrooms were determined using the Newton model and the Classical diffusion model. Activation energy (E _a ) values as determined using the Newton model were 22.58 and 20.48 kJ/mol for the whole and sliced mushrooms, respectively.     

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.     

Heavy metal contents and chemical compositions of atlantic (<Emphasis Type="Italic">Scomber scombrus</Emphasis>), blue (<Emphasis Type="Italic">Scomber australasicus</Emphasis>), and chub (<Emphasis Type="Italic">Scomber japonicus</Emphasis>) mackerel muscles

The heavy metal contents and chemical compositions among mackerel species were investigated. Atlantic and chub mackerel had higher mercury, but lower lead than blue mackerel. Chub mackerel had the highest crude fat (18.62%) and the lowest moisture (58.29%), whereas blue mackerel had the highest moisture (70.10%) and the lowest crude fat (2.89%). Atlantic and blue mackerel had total saturated fatty acids (SFA)>polyunsaturated fatty acids (PUFA)>monounsaturated fatty acids (MUFA), whereas chub mackerel had PUFA>SFA>MUFA. The main SFA of the 3 mackerels were 16:0 and the main PUFAs were 22:6n-3 and 20:5n-3. The total amino acid contents in Atlantic, blue, and chub mackerel were 190.63, 169.49, and 172.97 mg/g, respectively. The major amino acids of the 3 mackerels were glutamic acid, aspartic acid, lysine, and leucine.     

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.     

Antitumor activities of liposome-incorporated aqueous extracts of <Emphasis Type="Italic">Anodonta </Emphasis><Emphasis Type="Italic">woodiana</Emphasis> (Lea, 1834)

To acquire the effectiveness of oral treatments, aqueous extracts of Anodonta woodiana (HB) were incorporated into liposome and its antitumor activities evaluated in vivo. The HB-loaded liposomes (HBL) were prepared at a mean size of 14.85 μm by reverse-phase evaporation method. Referring to the maximum tolerated doses test, mice with orally administrated HBL, at a 3 g/kg body weight dosage, showed no obvious acute toxic sign. Furthermore, the tumoricidal activities of HBL against C26 murine colon carcinoma, Lewis lung tumor, human QGY hepatic carcinoma and MKN-45 gastric tumor were examined, respectively. In contrast to free HB, HBL possessed remarkable antitumor activity. Simultaneously, the effect of HBL was observed in a dose-dependent manner. For C26 murine colon carcinoma and Lewis lung tumor, the inhibitory ratios of HBL were 54.36 and 51.97% at a dose of 400 mg/kg, respectively. The suppression of tumor growth and the reduction in body weight were more pronounced in human QGY hepatic carcinoma and MKN-45 gastric tumors inoculated mice by treatment of HBL during 30 days. All these promising results implied that liposome-incorporated aqueous extracts of Anodonta woodiana had a more potential application as a natural antitumor and immunomodulator formulation.     

Tyramine reduction by tyrosine decarboxylase inhibitor in <Emphasis Type="Italic">Enterococcus faecium</Emphasis> for tyramine controlled <Emphasis Type="Italic">cheonggukjang</Emphasis>

This study was carried out to find a method to control tyrosine decarboxylase activity (TDC) of a strain of Enterococcus faecium capable of producing high levels of tyramine. To select a TDC inhibitor, enzyme assay was first performed using purified TDC enzyme and 0.1% of TDC inhibiting chemicals. When 0.23% of nicotinic acid was added, tyramine content (363 ug/mL) was lower than that of the control group (873 ug/mL). At the same time, bacterial growth was decreased 1 log cycle from 8.62 to 7.56 log CFU/mL. TDC expression level in E. faecium was measured by using RT-qPCR. Lower expression level (below 0.7) was observed after the addition of 0.23% nicotinic acid (in vitro). When cheonggukjang was manufactured with addition of nicotinic acid, tyramine contents were decreased from 698.67 to 117.27 mg/kg when the concentration of nicotinic acid added was increased from 0.10 to 0.30%. These results suggest that nicotinic acid could be used as an agent (TDC inhibitor) to reduce tyramine content in cheonggukjang.     

Improvement of <Emphasis Type="Italic">Candida parapsilosis</Emphasis> by genome shuffling for the efficient production of arabitol from <Emphasis Type="SmallCaps">l</Emphasis>-arabinose

Arabitol is used in the food industry as a low-calorie sweetener. It is produced by yeasts during the biotransformation process of l-arabinose. Genome shuffling was performed in Candida parapsilosis DSM 70125, an efficient producer of arabitol, to obtain fusants with improved arabitol production ability. Four mutants from the parental library were used for the first round of genome shuffling. The best fusants, GSI-1 and GSI-10A, were subjected to a second round of genome shuffling. Finally, two fusants, GSII-3 and GSII-16, produced concentrations of arabitol that were 50% higher than that of the wild-type strain during selection culture. Under the optimal conditions established for C. parapsilosis, the two fusants produced 11.83 and 11.75 g/L of arabitol and were approximately 15–16% more efficient than the wild-type strain. Flow cytometry analysis showed that the ploidy of the new strains did not change.     

Development and characterization of reconstituted hydrogel from <Emphasis Type="Italic">Aloe vera</Emphasis> (<Emphasis Type="Italic">Aloe barbadensis</Emphasis> Miller) powder

Structural and rheological characterization of reconstituted hydrogels developed from A. vera non-fibrous alcohol insoluble residue (NFAIR) powder using different methods [viz., shaking (S), heating-shaking (HS), and heating (H)] and concentrations (viz., 0.2–1.6 %, w/v) was carried out. Functional group distribution by FTIR spectroscopy and Congo red (CR) method revealed the presence of acetylated acemannan in A. vera powder. Dynamic oscillation studies of A. vera (NFAIR) fluids at all concentrations of 0.2–1.6 %, w/v, showed gel strength in the order of H > HS > S method. However, in H method, increase in concentration from 0.2 to 1.6 %, w/v showed the conformational transition from semi-diluted solution to weak gel nature. Rheological models described the effect of heating temperatures (HT); 30–90 °C, and times (Ht); 15–60 min on viscoelastic behavior in reconstituted A. vera fluids. The reconstituted A. vera hydrogel prepared with a concentration of 1.6 %, w/v using 50 °C (HT) and 30 min (Ht) condition showed a good agreement with the Power law (storage modulus, G′) and Weak gel model (complex modulus, G*) fitted data (R² > 0.94) resulting higher viscoelastic moduli intercepts; G′₀ (71.5 Pa s ^n′), G″₀ (33.5 Pa s ^n″), lower slopes; n′ (0.22), n″ (0.06), higher network strength (A _F , 121.3 Pa s^1/z ) and number of network (z, 5.3) values. The obtained results suggested that heating at 50 °C/30 min can develop aqueous weak gel networks of A. vera with enhanced gel strength which may be utilized as a novel gelling agent for wide variety of targeted applications in food and pharmaceutical sectors.     

Characterization of brewing microorganisms isolated from Korean traditional <Emphasis Type="Italic">nuruk</Emphasis> for <Emphasis Type="Italic">Cheongju</Emphasis> production

The objective of this study was to select and identify yeasts and molds isolated from traditional nuruk and to investigate their brewing characteristics for Cheongju production. The yeast strains Y190 (Accession ID-KACC 93251P), Y263 (Accession ID-KACC 93252P), and Y270 (Accession ID-KACC 93253P) showing high alcohol and flavor productivity were isolated and identified by phylogenetic inference based on an internal transcribed spacer 2 region sequence analysis. In addition, Aspergillus oryzae 83-10 (Accession ID-KACC 93254P) showing the highest enzyme activity was isolated. This study provides basic data for the production of Korean Cheongju and assesses the applicability of these three yeast strains and A. oryzae 83-10 isolated from traditional nuruk.     

Characterization of mannanase from <Emphasis Type="Italic">Bacillus</Emphasis> sp., a novel <Emphasis Type="Italic">Codium fragile</Emphasis> cell wall-degrading bacterium

Seaweeds are considered as a health food partly due to the polysaccharide composition of the cell wall. Because conventional extraction methods have low yields and lead to environmental pollution, enzymatic methods have been proposed. In this study, a new strain of Bacillus sp. was isolated from cattle feces that produced a mannanase, a polysaccharide-degrading enzyme active against the green seaweed Codium fragile. The purified 39-kDa mannanase exhibited maximum activity at 55 °C and pH 6.0, and maintained its catalytic activity stably at temperatures up to 60 °C and at a broad pH range (5.0–11.0). Enzymatic activity was slightly enhanced by Cu²⁺ and Na⁺ but strongly inhibited by Fe²⁺, Ag⁺, and EDTA. The mannanase showed the highest specificity to the inexpensive substrates such as konjac powder and locust bean gum, and efficiently released various manno-oligosaccharides. This novel mannanase can thus be applicable in the food, feed, and pulp industries.     

Modeling the Inactivation of <Emphasis Type="Italic">Listeria innocua</Emphasis> and <Emphasis Type="Italic">Escherichia coli</Emphasis> in Fresh-Cut Tomato Treated with Pulsed Light

The effectiveness of pulsed light (PL) treatments to inhibit microorganisms on fresh-cut tomatoes (Lycopersicon esculentum Mill., cv. Daniela) was investigated. Tomato slices inoculated with Escherichia coli or Listeria innocua were exposed to PL treatments (4, 6, or 8 J cm^?2 fluence) and kept cold at 4 °C for 20 days. L. innocua and E. coli counts, gases in the headspace of the containers (O₂ and CO₂), pH, titratable acidity, and soluble solid content were monitored throughout the cold storage. The PL treatments reduced significantly (p < 0.05) initial loads of both microbes. The effect of the PL fluence on the survival number of microoganisms was described by a log-linear model (R ² = 0.849–0.999). At any fixed time within the cold storing, the microbial counts for untreated samples were always higher than those cut tomatoes that had been previously PL-treated. The behavior of L. innocua and E. coli during the storage were well adjusted (R ² > 0.930) by Gompertzian models; the studied microorganisms exhibited different patterns during the storage period. On the other hand, O₂ and CO₂ partial pressures in containers with fresh-cut tomatoes were also significantly affected by PL treatments (p < 0.05). The highest PL fluence caused the greatest changes of O₂ and CO₂ contents. In addition, the application of PL triggered an acceleration of the O₂ consumption during the cold stage. PL treatments might be used to effectively extend the safety of fresh-cut tomatoes over 12 days of storage against E. coli and L. innocua growth.     

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.