Search results for "electrophoresis"
showing 10 items of 1007 documents
Three-dimensional crystallization of the light-harvesting complex from Mantoniella squamata (Prasinophyceae) requires an adequate purification proced…
1995
Abstract We present a new purification procedure for the light-harvesting complex of Mantoniella squamata whereupon three-dimensional crystallization succeeded. Previous purification methods were based on density centrifugations as the only separating principle. We have extended this preparation procedure by applying anion-exchange and molecular-sieve chromatography techniques. Purity and stability of the complex were proved by denaturing and non-denaturing polyacrylamide-gel electrophoresis, and spectroscopic measurements. With respect to contaminating lipids the purified pigment-protein complex was examined by thin-layer chromatography and the aggregation and/or oligomeric states were inv…
Recombinant laccase from Pediococcus acidilactici CECT 5930 with ability to degrade tyramine
2017
Biogenic amines degradation by bacterial laccases is little known, so we have cloned and heterologously expressed, in E. coli, a new laccase from Pediococcus acidilactici CECT 5930 (Lpa5930), a lactic acid bacterium commonly found in foods able to degrade tyramine. The recombinant enzyme has been characterized by physical and biochemical assays. Here we report the optimization of expression and purification procedures of this laccase. DNA encoding sequence of laccase from P. acidilactici was amplified by PCR and cloned into the expression plasmid pET28a for induction by isopropyl-β-D-thiogalactoipyranoside. Protein expression was performed in E. coli BL21(DE3) harboring pGro7 plasmid expres…
An integrated proteomic and metabolomic study to evaluate the effect of nucleus-cytoplasm interaction in a diploid citrus cybrid between sweet orange…
2018
Key message: Our results provide a comprehensive overview how the alloplasmic condition might lead to a significant improvement in citrus plant breeding, developing varieties more adaptable to a wide range of conditions. Abstract: Citrus cybrids resulting from somatic hybridization hold great potential in plant improvement. They represent effective products resulting from the transfer of organelle-encoded traits into cultivated varieties. In these cases, the plant coordinated array of physiological, biochemical, and molecular functions remains the result of integration among different signals, which derive from the compartmentalized genomes of nucleus, plastids and mitochondria. To dissect …
The shell matrix of the pulmonate land snail Helix aspersa maxima.
2012
12 pages; International audience; In mollusks, the shell mineralization process is controlled by an array of proteins, glycoproteins and polysaccharides that collectively constitute the shell matrix. In spite of numerous researches, the shell protein content of a limited number of model species has been investigated. This paper presents biochemical data on the common edible land snail Helix aspersa maxima, a model organism for ecotoxicological purposes, which has however been poorly investigated from a biomineralization viewpoint. The shell matrix of this species was extracted and analyzed biochemically for functional in vitro inhibition assay, for amino acid and monosaccharides composition…
Subcellular localization and purification of a p-hydroxyphenylpyruvate dioxygenase from cultured carrot cells and characterization of the correspondi…
1997
p-Hydroxyphenylpyruvate dioxygenase catalyses the transformation of p-hydroxyphenylpyruvate into homogentisate. In plants this enzyme has a crucial role because homogentisate is the aromatic precursor of all prenylquinones. Furthermore this enzyme was recently identified as the molecular target for new families of potent herbicides. In this study we examine precisely the localization of p-hydroxyphenylpyruvate dioxygenase activity within carrot cells. Our results provide evidence that, in cultured carrot cells, p-hydroxyphenylpyruvate dioxygenase is associated with the cytosol. Purification and SDS/PAGE analysis of this enzyme revealed that its activity is associated with a polypeptide of 4…
Impact of Pressurized Liquid Extraction and pH on Protein Yield, Changes in Molecular Size Distribution and Antioxidant Compounds Recovery from Spiru…
2021
The research aims to extract nutrients and bioactive compounds from spirulina using a non-toxic, environmentally friendly and efficient method—Pressurized Liquid Extraction (PLE). In this work, Response Surface Methodology (RSM)–Central Composite Design (CCD) was used to evaluate and optimize the extraction time (5–15 min), temperature (20–60 °C) and pH (4–10) during PLE extraction (103.4 bars). The multi-factor optimization results of the RSM-CCD showed that under the pressure of 103.4 bars, the optimal conditions to recover the highest content of bioactive compounds were 10 min, 40 °C and pH 4. Furthermore, the compounds and antioxidant capacity of PLE and non-pressurized extraction extra…
Preferential induction of 20S proteasome subunits during elicitation of plant defense reactions: towards the characterization of "plant defense prote…
2003
Plants have evolved efficient mechanisms to resist pathogens. The earliest defense response is the hypersensitive response (HR) considered as the main step leading to plant systemic acquired resistance (SAR) that protects the whole plant against a large spectrum of pathogens. We showed previously that elicitation of defense reactions in tobacco cells by cryptogein, a proteinaceous elicitor of plant defense reactions, leads to a rapid and differential accumulation of transcripts corresponding to genes encoding defense-induced (din) subunits of 20S proteasome: beta1din, alpha3din and alpha6din.Here, expression of these three subunits was investigated by Northern blotting and by Western blotti…
Removal of 2-butoxyethanol gaseous emissions by biotrickling filtration packed with polyurethane foam
2016
The removal of 2-butoxyethanol from gaseous emissions was studied using two biotrickling filters (BTF1 and BTF2) packed with polyurethane foam. Two different inoculum sources were used: a pure culture of Pseudomonas sp. BOE200 (BTF1) and activated sludge from a municipal wastewater treatment plant (BTF2). The bioreactors were operated at inlet loads (ILs) of 130 and 195 g m(-3) hour(-1) and at an empty bed residence time (EBRT) of 12.5s. Under an IL of ∼130 g m(-3) hour(-1), BTF1 presented higher elimination capacities (ECs) than BTF2, with average values of 106±7 and 68±8 g m(-3) hour(-1), respectively. However, differences in ECs between BTFs were decreased by reducing the irrigation inte…
A technical trick for studying proteomics in parallel to transcriptomics in symbiotic root-fungus interactions
2004
We have developed a protocol in which proteins and mRNA can be analyzed from single root samples. This experimental design was validated in arbuscular mycorrhiza by comparing the proteins profiles obtained with those from a classical protein extraction process. It is a step forward to make simultaneous proteome and transcriptiome profiling possible.
Proteomics as a way to identify extra-radicular fungal proteins from Glomus intraradices - RiT-DNA carrot root mycorrhizas
2004
To identify fungal proteins involved in the arbuscular mycorrhizal symbiosis, root-inducing transferred-DNA transformed roots of carrot (Daucus carota L.) were in vitro inoculated with Glomus intraradices. Proteins extracted from the extra-radical fungus were analysed by two-dimensional gel electrophoresis. A fungal reference map displaying 438 spots was set up. Four proteins, among the 14 selected for tandem mass spectrometry analysis, were identified including a NmrA-like protein, an oxido-reductase, a heat-shock protein and an ATP synthase beta mitochondrial precursor. The possible fungal origin of a MYK15-like protein found in mycorrhizal roots was further discussed. This is the first r…