Search results for "Molecular mass"
showing 10 items of 155 documents
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…
Characterization of a β-glucosidase isolated from an alpeorujo strain of Candida adriatica
2017
ABSTRACTA β-glucosidase-producing strain, Candida adriatica CECT13142, was isolated from olive oil wastes (alpeorujo) and identified by PCR/restriction fragment length polymorphism of the rDNA internal transcribed spacer and sequence analysis of the D1/D2 region of the 26S rRNA gene techniques. The enzyme was purified by sequential chromatography on DEAE-cellulose and Sephadex G-100. The relative molecular mass of the enzyme was estimated to be 50 kDa by SDS-PAGE. The hydrolytic activity of the β-glucosidase had an optimum pH of 8.2 and an optimum temperature of 40°C. The enzyme displayed high substrate specificity and high catalytic efficiency (Km 0.85 mM, Vmax 12.5 U/g of cells) for p-nit…
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…
Purification and characterization of geranyl diphosphate synthase from Vitis vinifera L. cv Muscat de Frontignant cell cultures
1993
A geranyl diphosphate synthase (EC 2.5.1.1), which catalyzes the formation of geranyl diphosphate from dimethylallyl diphosphate and isopentenyl diphosphate, was isolated from Vitis vinifera L. cv Muscat de Frontignan cell cultures. Purification of the enzyme was achieved successively by ammonium sulfate precipitation and chromatography on DEAE-Sephacel, hydroxylapatite, Mono Q, Phenyl Superose, Superose 12, and preparative nondenaturing polyacrylamide gels. The enzyme formed only geranyl diphosphate as a product. In all cases, neither neryl diphosphate, the cis isomer, nor farnesyl diphosphate was detected. The enzyme showed a native molecular mass of 68 [plus or minus] 5 kD as determined …
Identification of Novel Phosphopeptides After Simulated Digestion of αs2-casein by Tandem Mass Spectrometry
2006
Casein phosphopeptides (CPPs) are encrypted in αs1-, αs2-and β-casein (CN) and can be released by in vitro, in vivohydrolysis or food processing of dairy foods. Bovine αs2-CN contains two cluster sequences of anionic phosphoseryl and glutamyl residues SpSpSpEE in its structure (residues 8–12 and 56–63), which can modulate mineral bioavailability. In this study αs2-casein (αs2-CN) was subjected to simulated gastrointestinal digestion. CPPs released were sequenced by on-line reversed-phase high performance liquid chromatography coupled to electrospray ionisation tandem mass spectrometry (RP-HPLC-ESIMS/MS). Six novel αs2-CN derived CPPs, Three of them (αs2-CN(54–87)4P,αs2-CN(24–70)4P and αs2-…
Identification of a Peptide Produced by Bifidobacterium longum CECT 7210 with Antirotaviral Activity
2016
Rotavirus is one of the main causes of acute diarrhea and enteritis in infants. Currently, studies are underway to assess the use of probiotics to improve rotavirus vaccine protection. A previous work demonstrated that the probiotic strain Bifidobacterium longum subsp. infantis CECT 7210 is able to hinder rotavirus replication both in vitro and in vivo. The present study takes a systematic approach in order to identify the molecule directly involved in rotavirus inhibition. Supernatant protease digestions revealed both the proteinaceous nature of the active substance and the fact that the molecule responsible for inhibiting rotavirus replication is released to the supernatant. Following pur…
Null mutants of Candida albicans for cell-wall-related genes form fragile biofilms that display an almost identical extracellular matrix proteome.
2016
By two-dimensional gel electrophoresis (2-DE) and mass spectrometry, we have characterized the polypeptide species present in extracts obtained by 60% ethanol treatment of whole mature (48 h) biofilms formed by a reference strain (CAI4- URA3 ) and four Candida albicans null mutants for cell-wall-related genes ( ALG5, CSA1, MNN9 and PGA10) . Null mutants form fragile biofilms that appeared partially split and weakly attached to the substratum contrary to those produced by the reference strain. An almost identical, electrophoretic profile consisting of about 276 spots was visualized in all extracts examined. Proteomic analysis led to the identification of 131 polypeptides, corresponding to 86…
Disclosing diversity of exopolysaccharide-producing lactobacilli from Spanish natural ciders
2018
24 p.-2 fig.-2 tab.-1 fig. supl.-1 tab.supl.
Synthesis and solution properties of star-shaped poly(tert-butyl acrylate)
2000
A series of star polymers consisting of poly(tert-butyl acrylate) arms and an ethyleneglycol dimethacrylate (EGDMA) microgel core were synthesized using anionic polymerization. The effect of various parameters (precursor length, ratio [[EGDMA]/[Initiator], reaction time, and overall concentrations) on the average number of arms was investigated. Molecular weights were determined using GPC coupled with an online viscometer and MALLS. The exponents for the relation between intrinsic viscosity or radius of gyration and molecular weight, respectively, are extremely low, indicating that the dimensions of the star polymers only slightly increase with the number of arms. After a certain number of …
Die bestimmung von sedimentationskoeffizient, diffusions-koeffizient und molekulargewicht von α-amylasen nach der methode des aktivitätstransportes
1965
Abstract Sedimentation and diffusion coefficients determined by optical methods on pure (crystallized) enzymes are compared with those obtained by activity measurements on the unpurified enzymes. On performing the experiments under suitable conditions, results of the two methods are in good agreement. The calculation of the molecular weights of the unpurified enzymes is somewhat uncertain because the partial specific volume in most cases is not accurately known.