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Low Salt (low + salt)
Terms modified by Low Salt Selected AbstractsA comparison of the urea-induced unfolding of apoflavodoxin and flavodoxin from Desulfovibrio vulgarisFEBS JOURNAL, Issue 1 2002Brian Ó Nuallain The kinetics and thermodynamics of the urea-induced unfolding of flavodoxin and apoflavodoxin from Desulfovibrio vulgaris were investigated by measuring changes in flavin and protein fluorescence. The reaction of urea with flavodoxin is up to 5000 times slower than the reaction with the apoprotein (0.67 s,1 in 3 m urea in 25 mm sodium phosphate at 25 °C), and it results in the dissociation of FMN. The rate of unfolding of apoflavodoxin depends on the urea concentration, while the reaction with the holoprotein is independent of urea. The rates decrease in high salt with the greater effect occurring with apoprotein. The fluorescence changes fit two-state models for unfolding, but they do not exclude the possibility of intermediates. Calculation suggests that 21% and 30% of the amino-acid side chains become exposed to solvent during unfolding of flavodoxin and apoflavodoxin, respectively. The equilibrium unfolding curves move to greater concentrations of urea with increase of ionic strength. This effect is larger with phosphate than with chloride, and with apoflavodoxin than with flavodoxin. In low salt the conformational stability of the holoprotein is greater than that of apoflavodoxin, but in high salt the relative stabilities are reversed. It is calculated that two ions are released during unfolding of the apoprotein. It is concluded that the urea-dependent unfolding of flavodoxin from D. vulgaris occurs because apoprotein in equilibrium with FMN and holoprotein unfolds and shifts the equilibrium so that flavodoxin dissociates. Small changes in flavin fluorescence occur at low concentrations of urea and these may reflect binding of urea to the holoprotein. [source] COMBINATION WITH PLANT EXTRACTS IMPROVES THE INHIBITORY ACTION OF DIVERGICIN M35 AGAINST LISTERIA MONOCYTOGENESJOURNAL OF FOOD QUALITY, Issue 1 2008ABDEL-MAJEED ZOUHIR ABSTRACT The susceptibility of 11 strains of Listeria monocytogenes to divergicin M35, a bacteriocin produced by Carnobacterium divergens strain M35, and to aqueous extracts of garlic, onion, oregano, red chili and black pepper at 30 and 10C, was evaluated using a microdilution assay. The susceptibility of divergicin-resistant strains to combinations of these agents was also evaluated. Three strains were resistant to divergicin M35 (>500 µg/mL) at 30C but were more susceptible at 10C. Garlic gave the most inhibitory plant extract, followed by onion, while oregano, red chili and black pepper extracts were less active at both temperatures. Garlic extract and divergicin M35 combined or with other extracts increased inhibitory activity against the divergicin-resistant strains. The garlic/divergicin combination was the most effective at inhibiting these strains and was bactericidal at both temperatures. Log-phase cells were the most susceptible to the garlic/divergicin combination. Stationary-phase cells were much more resistant at both incubation temperatures. Furthermore, the effect of the garlic/divergicin combination at inhibiting divergicin-resistant L. monocytogenes in a food system was also studied using cold-smoked salmon as a food model. Results indicated that this combination could efficiently reduce the viability of L. monocytogenes in smoked salmon stored at 10C. PRACTICAL APPLICATIONS There is increasing popularity worldwide for chemical preservative-free, ready-to-eat and minimally processed seafood with low salt, fat and sugar content. Bacteriocins produced from lactic acid bacteria can have a potential application to prolong the shelf life of cold-smoked salmon. Also, plant and spice extracts have been shown to contain antibacterial substances with potential for application in foods. Thus, this research explores the combination of divergicin M35, a bacteriocin produced by Carnobacterium divergens strain M35, and aqueous extracts of garlic, onion, oregano, red chili and black pepper to inhibit Listeria monocytogenes and to prolong the shelf life of cold-smoked salmon. [source] Stabilization of human papillomavirus virus-like particles by non-ionic surfactantsJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 7 2005Li Shi Abstract Human papillomavirus (HPV) virus-like-particles (VLPs) produced by recombinant expression systems are promising vaccine candidates for prevention of cervical cancers as well as genital warts. At high protein concentrations, HPV VLPs, comprised of the viral capsid protein L1 and expressed and purified from yeast, are protected against detectable aggregation during preparation and storage by high concentrations of NaCl. At low protein concentrations, however, high salt concentration alone does not fully protect HPV VLPs from aggregation. Moreover, the analytical analysis of HPV VLPs proved to be a challenge due to surface adsorption of HPV VLPs to storage containers and cuvettes. The introduction of non-ionic surfactants into HPV VLP aqueous solutions provides significantly enhanced stabilization of HPV VLPs against aggregation upon exposure to low salt and protein concentration, as well as protection against surface adsorption and aggregation due to heat stress and physical agitation. The mechanism of non-ionic surfactant stabilization of HPV VLPs was extensively studied using polysorbate 80 (PS80) as a representative non-ionic surfactant. The results suggest that PS80 stabilizes HPV VLPs mainly by competing with the VLPs for various container surfaces and air/water interfaces. No appreciable binding of PS80 to intact HPV VLPs was observed although PS80 does bind to the denatured HPV L1 protein. Even in the presence of stabilizing level of PS80, however, an ionic strength dependence of HPV VLP stabilization against aggregation is observed indicating optimization of both salt and non-ionic surfactant levels is required for effective stabilization of HPV VLPs in solution. © 2005 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 94:1538,1551, 2005 [source] Guanine quadruplex formation by RNA/DNA hybrid analogs of Oxytricha telomere G4T4G4 fragmentBIOPOLYMERS, Issue 10 2008Jitka Vondru Abstract Using circular dichroism spectroscopy, gel electrophoresis, and ultraviolet absorption spectroscopy, we have studied quadruplex folding of RNA/DNA analogs of the Oxytricha telomere fragment, G4T4G4, which forms the well-known basket-type, antiparallel quadruplex. We have substituted riboguanines (g) for deoxyriboguanines (G) in the positions G1, G9, G4, and G12; these positions form the terminal tetrads of the G4T4G4 quadruplex and adopt syn, syn, anti, and anti glycosidic geometries, respectively. We show that substitution of a single sugar was able to change the quadruplex topology. With the exception of G4T4G3g, which adopted an antiparallel structure, all the RNA/DNA hybrid analogs formed parallel, bimolecular quadruplexes in concentrated solution at low salt. In dilute solutions (,0.1 mM nucleoside), the RNA/DNA hybrids substituted at positions 4 or 12 adopted antiparallel quadruplexes, which were especially stable in Na+ solutions. The hybrids substituted at positions 1 and 9 preferably formed parallel quadruplexes, which were more stable than the nonmodified G4T4G4 quadruplex in K+ solutions. Substitutions near the 3,end of the molecule affected folding more than substitutions near the 5,end. The ability to control quadruplex folding will allow further studies of biophysical and biological properties of the various folding topologies. © 2008 Wiley Periodicals, Inc. Biopolymers 89: 797,806, 2008. This article was originally published online as an accepted preprint. The "Published Online" date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com [source] Effects of ionic strength on lysozyme uptake rates in cation exchangers.BIOTECHNOLOGY & BIOENGINEERING, Issue 2 2005I: Uptake in SP Sepharose FF Fluorescence scanning confocal microscopy was used in parallel with batch uptake and breakthrough measurements of transport rates to study the effect of ionic strength on the uptake of lysozyme into SP Sepharose FF. In all cases the adsorption isotherms were near-rectangular. As described previously, the intraparticle profiles changed from slow-moving self-sharpening fronts at low salt concentration, to fast-moving diffuse profiles at high salt concentration, and batch uptake rates correspondingly increased with increasing salt concentration. Shrinking core and homogeneous diffusion frameworks were used successfully to obtain effective diffusivities for the low salt and high salt conditions, respectively. The prediction of column breakthrough was generally good using these frameworks, except for low-salt uptake results. In those cases, the compressibility of the stationary phase coupled with the shrinking core behavior appears to reduce the mass transfer rates at particle-particle contacts, leading to shallower breakthrough curves. In contrast, the fast uptake rates at high ionic strength appear to reduce the importance of mass transfer limitations at the particle contacts, but the confocal results do show a flow rate dependence on the uptake profiles, suggesting that external mass transfer becomes more limiting at high ionic strength. These results show that the complexity of behavior observable at the microscopic scale is directly manifested at the column scale and provides a phenomenological basis to interpret and predict column breakthrough. In addition, the results provide heuristics for the optimization of chromatographic conditions. © 2005 Wiley Periodicals, Inc. [source] Circular dichroism spectroscopy of conformers of (guanine + adenine) repeat strands of DNACHIRALITY, Issue 7 2003Iva Kejnovská Abstract (Guanine+adenine) strands of DNA are known to associate into guanine tetraplexes, homodimerize into parallel or antiparallel duplexes, and fold into a cooperatively melting single strand resembling the protein alpha helix. Using CD spectroscopy and other methods, we studied how this conformational polymorphism depended on the primary structure of DNA. The study showed that d(GGGA)5 and d(GGA)7 associated into homoduplexes at low salt or in the presence of LiCl but were prone to guanine tetraplex formation, especially in the presence of KCl. In addition, they yielded essentially the same CD spectrum in the presence of ethanol as observed with the ordered single strand of d(GA)10. Strands of d(GA)10, d(GGAA)5, d(GAA)7, and d(GAAA)5 associated into homoduplexes in both LiCl and KCl solutions, but not into guanine tetraplexes. d(GAAA)5 and d(GAA)7 further failed to form the single-stranded conformer in aqueous ethanol. Adenine protonation, however, stabilized the single-stranded conformer even in these adenine-rich fragments. The ordered single strands, homoduplexes as well as the guanine tetraplexes, all provided strikingly similar CD spectra, indicating that all of the conformers shared similar base stacking geometries. The increasing adenine content only decreased the conformer thermostability. Chirality 15:584,592, 2003. © 2003 Wiley-Liss, Inc. [source] | ||||||||||