Content and distribution of vicine,convicine and <Emphasis Type="SmallCaps">l</Emphasis>-DOPA during germination and seedling growth of two <Emphasis Type="Italic">Vicia faba</Emphasis> L. varieties

This work examined the content and distribution of the pyrimidine glucosides vicine and convicine, and the nonproteic amino acid l-DOPA in cotyledons and embryo axes along the germination and seedling growth of Vicia faba L. vars. Alameda and Brocal. The behaviour of these compounds was similar in both varieties. Vicine and convicine, implicated in favism disease, slowly declined in cotyledons. In embryo axis, vicine levels were sharply reduced and convicine amount was slightly increased as assay progressed. Total pyrimidine glucosides remained unchanged in the whole plant during the study. l-DOPA only appeared in the embryo axis of V. faba seeds and the highest content was observed in Brocal variety at 6 DAI (days after imbibition). The information provided in this study could be valuable for a possible role of embryo axis, rich in l-DOPA, for the treatment of Parkinson’s disease and also as a choice ingredient for functional foods or as nutraceutical.     

Starch,protein, glycoalkaloids,and <Emphasis Type="SmallCaps">l</Emphasis>-ascorbic acid content in tubers of genetically modified potato cv. Irga

Changes in contents of protein, starch, ash, l-ascorbic acid, and glycoalkaloids in tubers of 15 clones of cultivar Irga transformed with viral genome sequences in order to improve their resistance to a necrotic strain of potato virus Y (PVY^N) were investigated. The influence of modification type and year of cultivation on the contents of all chemical compounds examined for particular modified clones appeared to be irregular over the 3-year period of examination, and the clone of the best and stable content of chemical compounds was not found. Stable tendencies in the chemical compounds of tubers composed into groups and subgroups of clones representing the same modification type were not confirmed either. An essential stable decrease in glycoalkaloid content and an increase in starch content (compared with parental cultivar Irga) were found in almost all genetically modified clones. Glycoalkaloid content in green tubers of modified clones was usually (except three clones) higher than this for parental cultivar Irga. Correlation between glycoalkaloid content and tuber size for all, normal and green tubers of selected clones and parental cultivar Irga was examined as well.     

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.     

Lipase-catalyzed acylation of <Emphasis Type="SmallCaps">l</Emphasis>-carnitine with conjugated linoleic acid in [Bmim]PF<Subscript>6</Subscript> ionic liquid

Improving significantly the acylation reaction of l-carnitine with conjugated linoleic acid catalyzed by lipase needs to select an efficient and suitable reaction medium that is not only polar but also hydrophobic. [Bmim]PF₆ which is satisfied with the above two requirements was applied as the reaction medium. The optimal reaction conditions were: Novozyme 435 as the catalyst, 5:1 of molar ratio of fatty acid to l-carnitine, 40 g L⁻¹ lipase, 0.22 a_W of initial water activity, 125 g L⁻¹ molecular sieves (they were added within 0–3 h of reaction time), 60 °C of reaction temperature, 200 rpm of rotation speed, 32 h of reaction time. The overall conversion could reach 98.27%. The conversion in [Bmim]PF₆ was 2.13 and 1.56 times higher than that in acetonitrile and in solvent-free system, respectively. The lipase in the present work could be used eight times. [Bmim]PF₆ as the reaction medium was better than acetonitrile and solvent-free system.     

Production of biogenic amines by<Emphasis Type="Italic"> Morganella morganii</Emphasis>,<Emphasis Type="Italic"> Klebsiella pneumoniae</Emphasis> and<Emphasis Type="Italic"> Hafnia alvei</Emphasis> using a rapid HPLC method

The histidine decarboxylating activity and production of biogenic amines by Morganella morganii (NCIMB, 10466), Klebsiella pneumoniae (NCIMB, 673) and Hafnia alvei (NCIMB, 11999) were investigated using a rapid HPLC method. Derivatisation of the bacterial samples was carried out using benzoyl chloride. A gradient elution system was used for analysis with a mixture of acetonitrile and HPLC grade water. Bacterial strains not only produce histamine in histidine-enriched broth but also the other biogenic amines. The chromatographic results show that bacterial strains are also capable of producing spermine and spermidine in histidine-enriched broth. Bacterial ammonia production by all three strains was clearly detected since ammonia is generated during the degradation of histidine. The study demonstrates that the highest histamine production was obtained by Morganella morganii, followed by Klebsiella pneumoniae, and the lowest with the Hafnia alvei. Therefore, Morganella morganii and Klebsiella pneumoniae have strong histidine decarboxylase activity since they are prolific histamine-forming bacteria     

A novel strategy to improve the bioconversion of l-carnitine from crotonobetaine

Using [Bmim]PF₆ ionic liquid as reaction medium, l-carnitine was biosynthesized through hydration of crotonobetaine under the action of carnitine dehydratase. l-Carnitine was transformed into palmitoyl-l-carnitine by lipase, which was extracted with tert-amyl alcohol. With the removal of l-carnitine, biotransformation of crotonobetaine into l-carnitine was continued in the reversible hydration reaction. According to this study, the molar yield of total l-carnitine could attain 87.2% after the five-cycle intermittent extraction, which was higher than all previous reports.     

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.     

Antioxidative activity of<Emphasis Type="Italic"> Geranium macrorrhizum</Emphasis>

The composition of radical-scavenging compounds from Geranium macrorrhizum leaves was analyzed and the antioxidative activities of various extracts was determined. Seven compounds, namely gallic acid, ellagic acid, 4-galloyl quinic acid, the flavonoid quercetin and three of its glycosides, quercetin-3--glucopyranoside, quercetin-3--galactopyranoside and quercetin-4--glucopyranoside were isolated and identified in the various fractions. The radical-scavenging activity of the isolated compounds was measured using DPPH^· and ABTS^·+ scavenging assays and compared with the activity of rosmarinic acid. Quercetin-3-glucopyranoside and quercetin-3-galactopyranoside showed the highest antioxidative capacities. Antioxidative activities of all plant fractions were assessed by model system tests (superoxide anion and hydrogen peroxide scavenging) and by oxidation tests (-carotene bleaching, peroxide value, UV absorbance and headspace-gas chromatography hexanal determination). The ethanol-butanol fraction of the plant possessed the highest activity in most of the tests.     

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.     

Investigations of the natural microflora of poppy seeds (<Emphasis Type="Italic">Papaver somniferum</Emphasis>) and hazelnut kernels (<Emphasis Type="Italic">Corylus avellana</Emphasis>) including microbiological decomposition

Fatty seeds of Papaver somniferum and Corylus avellana undergo a rapid microbial degradation after being ground. Those bacteria and fungi which are mainly responsible for the microbial decay were identified, and the most important growth and death processes were documented using crucial indicator-organisms. Additionally, an aflatoxin-screening was carried out in order to assess the possible risk-potential of food intoxication. The acid value (indicator for free fatty acids) of poppy seeds and hazelnut kernels was determined during their fermentation in order to document the decomposition of triglycerides.In this study it could be proved that initially a natural decay of oil seeds is caused by bacteria, yeasts and mould fungi. After the bacteria died in the course of time, yeasts and mould fungi dominated the germ spectrum. Bacteria taking part in the degradation were identified as varieties of Staphylococcus xylosus, Enterobacteriaceae and coliforms. Yeasts were identified as Pichia burtonii, and the mould fungi are associated with the genus Alternaria.On account of the absence of the genus Aspergillus in the spectrum of mould fungi, no aflatoxin was produced.     

Antioxidant activity of <Emphasis Type="Italic">Dioscorea batatas</Emphasis> Decne glycoprotein

Dioscorea batatas Decne (DBD) has been traditionally used as folk medicine and health food in Korea. One glycoprotein was isolated from DBD and confirmed to have 30 kDa molecular weight. The DBD glycoprotein was tested its antioxidative activity and characterized in various chemical conditions. The DBD glycoprotein has the optimal free radical scavenging activities in acidic and neutral pH and up to 85 °C. In the M²⁺ ions (Ca²⁺, Mn²⁺, and Mg²⁺) in the presence of EDTA, the activities of DBD glycoprotein reduced, compared to DBD glycoprotein alone without metal ion. Interestingly, the results in this study indicated that the activities of DBD glycoprotein do not depend on the presence of EDTA. Interestingly, when DBD glycoprotein was treated with deactivation agents (pronase E or NaIO₄), scavenging activity of DBD glycoprotein was decreased. The anti-oxidative effects of DBD glycoprotein on hydroxyl radicals in cell-free system revealed, and the DBD glycoprotein has remarkable scavenging effects on hydroxyl radicals generated by G/GO. Furthermore, the results in this study showed that the DBD glycoprotein (200 μg/ml) significantly inhibits intracellular ROS amounts and protects from cytotoxicity in primary mouse splenocyte culture treated with GO (30 mU/ml). Therefore, we speculate that DBD glycoprotein has an antioxidative potential as one of natural antioxidants.     

Influence of<Emphasis Type="Italic"> Lactobacillus plantarum</Emphasis> on<Emphasis Type="Italic"> Staphylococcus aureus</Emphasis> growth in a fresh vegetable model system

The influence of a Lactobacillus plantarum (B₄) on the growth of Staphylococcus aureus (Sa₄) was verified by impedometric methods in a suitable model reproducing the characteristics of fresh vegetables. The inoculum size of the single strains and their growth temperature were varied according to a Central Composite Design. The results obtained via statistical analysis showed that the temperature affected the growth of both S. aureus and L. plantarum strains. The pathogenic strain, independently of its inoculum size, was inhibited by L. plantarum at all the tested temperatures. A proper combination of specific lactic acid bacteria and storage temperature should improve the safety of the vegetable products.     

Prevalence,genetic diversity,and antibiotic susceptibility of <Emphasis Type="Italic">Cronobacter</Emphasis> spp. (<Emphasis Type="Italic">Enterobacter sakazakii</Emphasis>) isolated from <Emphasis Type="Italic">Sunshik</Emphasis>, its ingredients and soils

The prevalence, genetic diversity, and antibiotic susceptibility of Cronobacter species (Enterobacter sakazakii) isolated from sunshik products, its ingredients, and root vegetable farm’s soils were investigated to analyze main reservoirs and contaminated sources of Cronobacter spp. Cronobacter spp. was isolated from 9 of 15 sunshik products, 26 of 72 its ingredients, and 2 of 39 soils. The root vegetables such as sweet potato and carrot showed higher contamination rate (70%) than the other sunshik ingredients. All isolates showed 929 bp band amplified from 16S rRNA and resistant to ampicillin, amoxicillin/clavulanic acid, and cefazolin. All isolates showed diverse random-amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) band patterns. However, 3 cases of RAPD banding patterns between clinical strains and isolates from sunshik products and root vegetables (yam, carrot) were related with similarities level of 80%. These studies indicated that root vegetables can be an important contamination source of Cronobacter spp. in sunshik products. Thus, the preparation of root vegetables for manufacturing sunshik products used as a weaning diet was handled with more care than the other sunshik ingredients.     

Modeling high pressure inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> in whole milk

The survival curves of Escherichia coli and Listeria innocua inactivated by high hydrostatic pressure (HHP) were obtained at room temperature (∼22 °C) and at five pressure levels (400, 450, 500, 550 and 600 MPa) in whole milk. These curves were described by the Weibull model and parameters of this model were reduced from two to one with slight loss of goodness-of-fit. The logarithm of the time constant parameter (δ) of the reduced Weibull model was described with respect to high pressure (P). This approach can be used to define a z _p value analogous to the modeling of the classical D value (increase in pressure that results in one log unit decrease of δ values). The development of accurate survival models under high pressure, as presented here, can be very beneficial to food industry for designing, evaluating and optimizing HHP processes as a new preservation technology.     

Novel anticoagulant compound from fermented red alga <Emphasis Type="Italic">Pachymeniopsis elliptica</Emphasis>

Natural fermentation was tested as a method of releasing active compounds during screening for potential anticoagulant activity in three types of algae (Pachymeniopsis elliptica, Sargassum horneri, and Ulva pertusa). Freeze dried algae samples (2.5 g) were fermented by adding 75 g of sugar and 500 mL of water and thereafter kept at room temperature (25 °C) for 3 months. Activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) were measured every 2 weeks for 3 months to determine the optimum time for the highest activity. Fermented P. elliptica, (which had the highest activity) was subjected to anion exchange chromatography (DEAE-cellulose) and sepharose 4B gel permeation chromatography. The purified sample was analyzed by agarose-gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) to confirm the purification and to determine the molecular mass, respectively. The 360 μg/mL of purified compound (Mwt > 500,000 Da) had both APTT and PT activities (>1,000 s). However, at the concentrations of 180 μg/mL, purified compound and heparin showed 540 and >1,000 s APTT activity, respectively. Though, the purified compound of P. elliptica considered as a weaker anticoagulant than heparin, this purified anticoagulant polysaccharide could be considered as a good alternative source as an anticoagulant. Moreover, the technique of fermentation is an inexpensive and feasible, this purified anticoagulant polysaccharide compound could be used in pharmaceutical and biomedical industry. Further investigations need to be performed to determine the mechanism of this novel anticoagulant compound. The authors Prashani Mudika Ekanayake and Chamilani Nikapitiya contributed equally to this work.     

Genetic typification by pulsed-field gel electrophoresis (PFGE) and randomly amplified polymorphic DNA (RAPD) of wild <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis> and <Emphasis Type="Italic">Oenococcus oeni</Emphasis> wine strains

Genetic typification of 120 bacterial isolates of Lactobacillus plantarum and Oenococcus oeni from different Rioja musts and wines was performed by numerical analysis of pulsed-field gel electrophoresis (PFGE) patterns with endonuclease SfiI, and 46 of them were also studied by randomly amplified polymorphic DNA (RAPD)-PCR. A comparative study of both typification methods applied to L. plantarum and O. oeni oenological strains was performed. Bacterial species was determined both by biochemical identification methods and by specific PCR analysis. A wide variety of restriction digest patterns were detected by PFGE among L. plantarum strains (36 unrelated patterns and one closely related pattern, out of 48 isolates), as well as among O. oeni strains (18 unrelated patterns out of 72 isolates). PFGE was shown to be a suitable method for strain differentiation and to determine which strains are present in wine fermentations, with a discriminatory power to type L. plantarum and O. oeni strains higher than that of RAPD-PCR.     

Efficiency of pulse pressure treatment for inactivation of <Emphasis Type="Italic">Escherichia coli</Emphasis> and <Emphasis Type="Italic">Listeria innocua</Emphasis> in whole milk

Escherichia coli ATCC 11775 and Listeria innocua ATCC 33090 in whole milk were inactivated by single- and multi-pulsed (up to 10 pulses) high hydrostatic pressure (HHP) treatments. Both bacteria showed similar resistance to single- and multi-pulsed HHP. The efficiency of pulsed pressure treatment depends on the combination of holding time of each pulse and number of pulses. It was observed that multi-pulsed pressure treatment instead of traditional single-pulsed HHP could be used to pasteurize milk at a lower pressure level. Nevertheless, an optimization is necessary between the pulse holding time, number of pulses, and pressure levels to reach the desirable log-reduction of microorganisms compatible with industrial application. This study was partly presented in Joint 21st AIRAPT and 45th EHPRG International Conference on High Pressure Science and Technology held September 17–21, 2007 in Catania (Italy).     

<Emphasis Type="Italic">Chromohalobacter beijerinckii</Emphasis>: a psychrophilic,extremely halotolerant and enzymatically active microbe from salted food with the capacity for biogenic amine production

The species Chromohalobacter beijerinckii comb. nov. was recently proposed in order to accommodate strain Pseudomonas beijerinckii Hof 1935, which was found in salted beans, and a novel isolate, strain 3b, which was obtained from salted herrings of the Baltic sea. The species is psychrophilic, growing from 5 to 42 °C and highly salt-tolerant, growing between 0.5 and 25% NaCl. Of interest to food microbiology are several properties, which are described here, such as the enzymatic activity of Ch. beijerinckii strain 3b against fish protein, cod liver oil and butter. In addition, the strain possesses decarboxylases of lysine, tyrosine and histidine and can thus be a producer of biogenic amines, mainly histamine and tyramine. In media containing 10% NaCl, Ch. beijerinckii strain 3b is resistant to the antibiotics bacitracin, chloramphenicol, streptomycin, tetracycline and novobiocin. The potential impact of these properties, especially of the biogenic amine content in salted food, which were not known so far from other species of the genus Chromohalobacter, is discussed.     

Properties of a fibrinolytic enzyme secreted by <Emphasis Type="Italic">Bacillus subtilis</Emphasis> JS2 isolated from saeu (small shrimp) <Emphasis Type="Italic">jeotgal</Emphasis>

Bacillus species were screened to be used as starters for jeotgals, salted and fermented Korean sea foods. A strain, JS2, showing strong fibrinolytic activity was isolated from saeu (small shrimp) jeotgal, and identified as Bacillus subtilis. Bacillus subtilis JS2 grew well at 20% (w/v) NaCl concentration. SDS-PAGE of culture supernatant from JS2 showed 3 major bands of 27, 29, and 60 kDa in size. Fibrin zymography showed that the 27 kDa band was the major fibrinolytic protein. The gene, aprEJS2, was cloned and introduced into B. subtilis WB600 using pHY300PLK. A B. subtilis transformant harboring pHYJS2 showed higher fibrinolytic activity than B. subtilis JS2. aprEJS2 was overexpressed in Escherichia coli BL21 (DE3). The optimum pH and temperature for AprEJS2 were pH 8.0 and 40 °C, respectively. Km and V_max values were determined. AprEJS2 has strong α-fibrinogenase activity and moderate β-fibrinogenase activity.