Search results for " ELECTROPHORESIS"
showing 10 items of 480 documents
Characterization of cell wall proteins from yeast and mycelial cells of Candida albicans by labelling with biotin: Comparison with other techniques
1992
Candida albicans ATCC 26555 blastoconidia and blastoconidia bearing germ tubes were metabolically labelled by incubating the cells with 14C-labelled protein hydrolysate and were subsequently tagged with biotin. Double-labelled (radioactive and biotinylated) cell wall proteins and glycoproteins were extracted from intact cells of both growth forms by treatment with 2-mercaptoethanol (beta ME) and with beta-glucanases (Zymolyase) after treatment with beta ME. The beta ME- and Zymolyase-extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blotted (immunoblotted) to nitrocellulose paper. Polyacrylamide gels were stained with Coomassie blue and process…
Salmon (Salmo salar) side streams as a bioresource to obtain potential antioxidant peptides after applying pressurized liquid extraction (PLE)
2021
The pressurized liquid extraction (PLE) technique was used to obtain protein extracts with antioxidant capacity from salmon muscle remains, heads, viscera, skin, and tailfins. A protein recovery percentage ≈28% was obtained for all samples except for viscera, which was ≈92%. These values represented an increase of 1.5–4.8-fold compared to stirring extraction (control). Different SDS-PAGE profiles in control and PLE extracts revealed that extraction conditions affected the protein molecular weight distribution of the obtained extracts. Both TEAC (Trolox equivalent antioxidant capacity) and ORAC (oxygen radical antioxidant capacity) assays showed an outstanding antioxidant activity for viscer…
Accelerated Solvent Extraction and Pulsed Electric Fields for Valorization of Rainbow Trout (Oncorhynchus mykiss) and Sole (Dover sole) By-Products: …
2021
Fishery by-products are rich in biologically active substances and the use of green and efficient extraction methods to recover these high-added-value compounds is of particular importance. In this study, head, skin and viscera of rainbow trout and sole were used as the target matrices and accelerated solvent extraction (ASE) (45–55 °C, 15 min, pH 5.2–6.8, 103.4 bars) and pulsed electric fields (PEF) (1–3 kV/cm, 123–300 kJ/kg, 15–24 h) were applied as extraction technologies. The results showed that ASE and PEF significantly increased the protein extract efficiency of the fish by-products (p < 0.05) by up to 80%. SDS-PAGE results showed that ASE and PEF treatments changed the molecular size…
Antioxidant capacity of trans -resveratrol dietary supplements alone or combined with the mycotoxin beauvericin
2017
Trans-resveratrol (trans-RSV) is a polyphenol with multiples biological properties, such as anti-inflammatory, antioxidant, anti-aging, anti-diabetic, and antiplatelet. It occurs naturally in grapes and derivate, peanuts and berries. Beauvericin (BEA) is a mycotoxin present in cereals that produces cytotoxicity, intracellular reactive oxygen species and lipid peroxidation. The general objective of this research was to evaluate whether trans-RSV could be used as a good polyphenol against damages produced by BEA. Because trans-RSV can be ingested through dietary supplements, to reach this goal, the following specific objectives were proposed: to determine a) the trans-RSV content in different…
Preliminary characterization of the material released to the culture medium by Candida albicans yeast and mycelial cells.
1995
Culture filtrate concentrates were obtained from Candida albicans yeast and mycelial cells grown in the presence of 14C-protein hydrolysate for radioactive labeling of cellular polypeptides. Both growth forms released to the medium minor but significant amounts of proteinaceous materials. The analysis of culture filtrate concentrates by means of SDS-polyacrylamide gel electrophoresis and fluorography revealed a similar and complex electrophoretic pattern, though some qualitative and quantitative differences between samples obtained from yeast and mycelial cells were observed. Materials released, mostly composed of mannoproteins as shown by their affinity towards concanavalin A, presented (i…
Glutamine synthetases of green and etiolated leaves ofSinapis alba : Evidence of the identity of the respective enzyme proteins.
1989
Studies on the glutamine synthetases (GS, EC 6.3.1.2) of green (GS2) and etiolated leaves (GSet) ofSinapis alba L. (cv. Steinacher) revealed striking similarities between the respective enzyme proteins. The enzymes showed corresponding chromatographic properties, both on dimethylaminoethyl-Sephacel and on hydroxylapatite columns. The purified GS proteins were also identical with regard to the molecular weight of their subunits. Isoelectrofocusing of pure GSet yielded two distinct polypeptide bands in the pH 5.6 region of the gels. This pattern corresponded to the two strong bands of GS2. Two charge variants of GS polypeptides could be detected by Western-blot analysis of the soluble protein…
UV-induced cross-linking of Tet repressor to DNA containing tet operator sequences and 8-azidoadenines.
1990
The synthesis of 8-azido-2'-deoxyadenosine-5'-triphosphate is described. The photoreactive dATP analog was characterized by thin layer chromatography, proton resonance spectroscopy, infrared spectroscopy and UV spectroscopy. Its photolysis upon UV irradiation was studied. After incorporation of this dATP analog into DNA containing the tet operator sequence the investigation of the interactions between tet operator DNA and Tet repressor protein by UV photocross-linking becomes possible. Photocross-linking of protein to DNA was demonstrated by the reduced migration of the DNA in SDS polyacrylamide gel electrophoresis. Addition of the inducer tetracycline prior to UV irradiation significantly …
Comparison of pulsed-field gel electrophoresis and whole-genome-sequencing-based typing confirms the accuracy of pulsed-field gel electrophoresis for…
2020
Summary Aim To determine whether pulsed-field gel electrophoresis (PFGE) accurately recognizes isolates belonging to clusters defined by techniques based on whole-genome sequencing (WGS) using Pseudomonas aeruginosa as a model. Methods We selected 65 isolates of ST395 P. aeruginosa isolated in seven European hospitals between 1998 and 2012. Isolates were typed by PFGE and sequenced by WGS. A core genome multi-locus sequence typing (cgMLST) analysis based on 3831 genes was performed with a homemade pipeline. Findings PFGE identified eight pulsotypes and cgMLST differentiated nine clusters and nine singletons. Five cgMLST clusters and pulsotypes (31/65 isolates) coincided perfectly. Isolates …
Shifts in size, genetic structure and activity of the soil denitrifier community by nematode grazing
2010
International audience; Bacterial-feeding nematodes represent an important driver of the soil microbial activity and diversity. This study aimed at characterizing the impact of nematode grazing on a model functional bacterial guild involved in N-cycling, the denitrifiers. Bacterial-feeding nematodes (Cephalobus pseudoparvus) were inoculated into soil microcosms whose indigenous nematofauna had previously been removed. The size, genetic structure and activity of the soil denitrifier community were characterized 15 and 45 days after nematodes inoculation using quantitative PCR of the nirK, nirS and nosZ denitrification genes, fingerprinting of the nirK and nirS genes and denitrification enzym…
Analytical methods to determine cocaine contamination of banknotes from around the world
2008
Abstract The presence of cocaine in a significant number of UK pounds (Xxxx), Euros (€) and North American banknotes ($) in general circulation requires appropriate tools to do determinations. This article discusses the-state-of-the-art in the analysis of cocaine on banknotes. We summarize the usual extraction methods of currency samples and compare them, especially with respect to avoiding sample damage. We critically discuss analytical methods, namely gas chromatography (GC) and liquid chromatography (LC), capillary electrophoresis (CE), immunoassay, thermal desorption tandem mass spectrometry (TD-MS2) and ion-mobility spectrometry (IMS). We also review cocaine levels on banknotes around …