The aim of the study was to broadly determine the biological activities of purple potato ethanolic extract of the Blue Congo variety (BCE). The antioxidant activity of BCE was determined in relation to liposome membranes, and peroxidation was induced by UVB and AAPH. To clarify the antioxidant activity of BCE, we investigated its interactions with hydrophilic and hydrophobic regions of a membrane using fluorimetric and FTIR methods. Next, we investigated the cytotoxicity and pro-apoptotic activities of BCE in two human colon cancer cell lines (HT-29 and Caco-2) and in normal cells (IPEC-J2). In addition, the ability to inhibit enzymes that are involved in pro-inflammatory reactions was examined. Furthermore, BCE interactions with serum albumin and plasmid DNA were investigated using steady state fluorescence spectroscopy and a single molecule fluorescence technique (TCSPC-FCS). We proved that BCE effectively protects lipid membranes against the process of peroxidation and successfully inhibits the cyclooxygenase and lipoxygenase enzymes. Furthermore, it interacts with the hydrophilic and hydrophobic parts of lipid membranes as well as with albumin and plasmid DNA. It was observed that BCE is more cytotoxic against colon cancer cell lines than normal IPEC-J2 cells; it also induces apoptosis in cancer cell lines, but does not induce cell death in normal cells.
- MeSH
- albuminy MeSH
- antioxidancia chemie farmakologie MeSH
- fytogenní protinádorové látky chemie farmakologie MeSH
- inhibitory cyklooxygenasy chemie farmakologie MeSH
- inhibitory lipoxygenas chemie farmakologie MeSH
- lidé MeSH
- lipidy chemie MeSH
- liposomy MeSH
- nádorové buněčné linie MeSH
- plazmidy MeSH
- reaktivní formy kyslíku MeSH
- rostlinné extrakty chemie farmakologie MeSH
- sérový albumin chemie metabolismus MeSH
- Solanum tuberosum chemie MeSH
- vazba proteinů MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Potato proteins are well known for their nutritional, emulsifying, foaming, gel forming or antioxidant properties that all make from them valuable protein source for food industry. Antifungal, antimicrobial and also antiviral properties, described for potato proteins in the review, enrich the possibilities of potato protein usage. Potato proteins were divided into patatin, protease inhibitors and fraction of other proteins that also included, besides others, proteins involved in potato defence physiology. All these proteins groups provide proteins and peptides with antifungal and/or antimicrobial actions. Patatins, obtained from cultivars with resistance to Phytophthora infestans, were able to inhibit spore germination of this pathogen. Protease inhibitors represent the structurally heterogeneous group with broad range of antifungal and antimicrobial activities. Potato protease inhibitors I and II reduced the growth of Phytophthora infestans, Rhizoctonia solani and Botrytis cinerea or of the fungi of Fusarium genus. Members of Kunitz family (proteins Potide-G, AFP-J, Potamin-1 or PG-2) were able to inhibit serious pathogens such as Staphylococcus aureus, Listeria monocytogenes, Escherichia coli or Candida albicans. Potato snakins, defensins and pseudothionins are discussed for their ability to inhibit serious potato fungi as well as bacterial pathogens. Potato proteins with the ability to inhibit growth of pathogens were used for developing of pathogen-resistant transgenic plants for crop improvement. Incorporation of potato antifungal and antimicrobial proteins in feed and food products or food packages for elimination of hygienically risk pathogens brings new possibility of potato protein usage.
- MeSH
- antibakteriální látky chemie farmakologie MeSH
- antifungální látky chemie farmakologie MeSH
- Bacteria účinky léků MeSH
- Candida albicans účinky léků MeSH
- houby účinky léků MeSH
- karboxylesterhydrolasy chemie farmakologie MeSH
- Listeria monocytogenes účinky léků MeSH
- peptidy chemie farmakologie MeSH
- Phytophthora účinky léků MeSH
- průmyslové fungicidy chemie farmakologie MeSH
- rostlinné proteiny chemie farmakologie MeSH
- Solanum tuberosum chemie MeSH
- Staphylococcus aureus účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Proteins were obtained from effluent of a starch manufacture by using different isolation temperatures (40, 60, 80, and 100 °C). The proteins, remaining in effluent after treatment of potato juice at 80 and 100 °C differed significantly in composition and in structural stability as well as in trypsin inhibitory and antifungal activities in comparison with the variants of 40 and 60 °C. The protein samples of 80 °C exhibited the highest antifungal activity and its average value of IC50 against five strains of two Fusarium species was determined in average at 0.18 mg ml-1. The 80 °C protein samples consisted predominantly of low-molecular proteins (7-17 kDa) identified as potato tuber protease inhibitors I and II. Predominantly, protease inhibitors II were identified for the protein samples obtained by 100 °C and here we identified 7 spots in comparison with 12 identified for the 80 °C samples. Samples of 40 and 60 °C with low antifungal activities represent high variability of detected and identified proteins. We identified various representatives of aspartic, cysteine, and serine protease inhibitors in both types of samples. These samples also contained Kunitz-type protease inhibitors that were not found in the 80 and 100 °C samples which documented thermal unstableness of Kunitz-type protease inhibitors. Functional stability at high temperatures and antifungal activity of isolated potato protease inhibitors I and II support the potential of this fraction usage in food, feed, pharmaceutical, or agricultural industry and offer new products for starch manufactures. At the same time, utilization of the stable protein fraction of waste deproteinized potato water promotes exploitation of potato starch production resources.
- MeSH
- 2D gelová elektroforéza MeSH
- antifungální látky farmakologie MeSH
- Fusarium účinky léků MeSH
- hlízy rostlin chemie MeSH
- mikrobiální testy citlivosti MeSH
- proteomika MeSH
- rostlinné proteiny chemie izolace a purifikace farmakologie MeSH
- škrob MeSH
- Solanum tuberosum chemie MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- stabilita proteinů MeSH
- tandemová hmotnostní spektrometrie MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
The objective of our study was to assess the extent of accumulation of cadmium, lead and zinc in potato tubers depending on the concentration of these heavy metals in soil and to evaluate the resistance of 11 cultivars of potato cultivated in 5 localities of the Slovakia against input of these heavy metals into the consumption part of potato. Contents of Cd (Pb, Zn) in soil were 0.94-2.54 (18.03-24.90, 35.71-72.40) mg/kg in soil extract by aqua regia and 0.030-0.188 (0.149-0.356, 0.052-0.238) mg/kg in soil extract by NH4NO3. The contents of Cd, Pb, and Zn were determined in potatoes in extracts of freeze-dried samples and expressed in mg/kg of fresh matter (FM). Determined contents of heavy metals were in the range of ND-0.058 mg Cd/kg FM, 0.020-0.630 mg Pb/kg FM, 1.836-3.457 mg Zn/kg FM, resp. The statistically significant correlation between heavy metal content in soil and its content in potato tubers were confirmed only: cv. Laura - Spissky Stvrtok (Cd), cv. Red Anna - Odorin (Pb) and Marabel, Red Anna - Odorin, cv. Marabel - Belusa, cv. Volumia - Imel (Zn).
- MeSH
- hlízy rostlin chemie růst a vývoj metabolismus MeSH
- kadmium analýza metabolismus MeSH
- látky znečišťující půdu analýza metabolismus MeSH
- olovo analýza metabolismus MeSH
- Solanum tuberosum chemie růst a vývoj metabolismus MeSH
- zinek analýza metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The analytical performance and evaluation of a kit-based ELISA for the determination of acrylamide in fried potato and corn chip samples are described. The sample homogenate is subjected to clean-up using SPE, followed by analyte derivatization and ELISA detection. Accuracy, precision and linearity of the ELISA procedure have been validated using spiked samples. Analytical recovery ranged from 91.8% to 96.0% with coefficients of variation below 15%. Good linearity over a wide range of dilution and minimal assay drift was observed within a microtiter plate. IC50 value of the calibration curve was 110 ng/mL, with the limit of detection about 5 ng/mL and dynamic range from 10 to 1000 ng/mL. The high specificity of the ELISA was demonstrated by cross-reactivity study using 11 potential cross-reactants. A good correlation between the results obtained from the ELISA and GC-MS within the concentration range 120-1500 μg/kg was found in the chip samples (r=0.992, n=120). The data demonstrate that the evaluated and validated ELISA has a potential utility in a quick, simple and reliable acrylamide screening analysis for the medium- and large-sized food companies, as well as for residue laboratories and the food industry dealing with improving the chemical safety of foods available to the consumer.
- MeSH
- akrylamid analýza MeSH
- analýza potravin metody MeSH
- ELISA metody MeSH
- kontaminace potravin analýza MeSH
- kukuřice setá chemie MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- reprodukovatelnost výsledků MeSH
- Solanum tuberosum chemie MeSH
- vaření metody MeSH
- vysoká teplota MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Application of Tris-N-[Tris(hydroxymethyl)methyl]glycine gels for 2DE is hampered by formation of mixed CHAPS-SDS micelles resulting in typical swirling pattern in the low mass range, which makes reliable quantitative and qualitative gel evaluation impossible. Modification of 2DE strip equilibration procedure prevented the direct interaction between both detergents during equilibration process, thus substantially improving gel separation.
Biochemical characteristics of patatin proteins purified by ion-exchange and affinity chromatography from tubers of 20 potato cultivars were studied to evaluate their genotype differences with respect to utility groups, table potato cultivars (TPCs) and processing potato cultivars (PPCs). Both groups of cultivars showed similar values of protein content in dry matter (3.98-7.39%) and of patatin relative abundance (5.40-35.40%). Three mass levels (∼40.6, 41.8, and 42.9 kDa) of purified patatins were found by MALDI-TOF MS within all cultivars. Differences among mass levels corresponding with the mass of sugar antenna (∼1.2 kDa) confirmed the previous concept of different glycosylation extentsin patatin proteins. It was showed that the individual types of patatin varying in their masses occur in the patatin family in a ratio specific for each of the cultivars, with the lowest mass type being the major one. Electrophoretic analyses demonstrated wide cultivar variability in number of patatin forms. Especially 2D-PAGE showed 17-23 detected protein spots independently on the utility group. Specific lipid acyl hydrolase (LAH) activity of purified patatins from the individual tested cultivars varied between 0.92 and 5.46 μmol/(min mg). Patatin samples within most of the TPCs exhibited higher values of specific LAH activity than samples of PPCs. It may be supposed that individual patatin forms do not have similar physiological roles.
- MeSH
- druhová specificita MeSH
- glykosylace MeSH
- hlízy rostlin chemie MeSH
- karboxylesterhydrolasy chemie izolace a purifikace metabolismus MeSH
- molekulová hmotnost MeSH
- protein - isoformy chemie izolace a purifikace metabolismus MeSH
- rostlinné proteiny analýza chemie izolace a purifikace metabolismus MeSH
- Solanum tuberosum chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
