The influence of bovine serum albumin on β-lactoglobulin denaturation, aggregation and gelation

peer-reviewedThe effect of bovine serum albumin (BSA) on the heat-induced denaturation, aggregation and subsequent acid-induced gelation of β-lactoglobulin (β-lg) was investigated in this work. Changes in the denaturation kinetics of β-lg during heating at 78 °C were determined by monitoring the disappearance of the native protein by reverse-phase chromatography. Replacing β-lg with increasing amounts of BSA, while keeping the total protein concentration constant at 5% (w/w), significantly increased the denaturation rate of β-lg from 2.57±0.30×10−3(g L−1)(1−n)s−1 to 5.07±0.72×10−3(g L−1)(1−n)s−1 (β-lg: BSA ratio of 3:1 w/w). The reaction order for β-lg was 1.40±0.09. Partial replacement of β-lg with BSA (β-lg: BSA ratio of 3:1 w/w) significantly increased the reaction order to 1.67±0.13. Heat-induced aggregates between β-lg and BSA were studied by dynamic light scattering, two-dimensional electrophoresis and size exclusion chromatography. The partial replacement of β-lg with BSA significantly changed the gelling properties of the acid-induced gels. A rapid rate of acidification resulted in a significant decrease, while a slow acidification rate resulted in a significant increase in gel strength. Size exclusion chromatography demonstrated that intermolecular disulphide bond formation occurred during both heat-induced denaturation/aggregation and subsequent acid-induced gelation. Results clearly indicate that BSA contributed to the formation of these disulphide bonds.This work was funded under the Food Institutional Research Measure (FIRM) of the National Development Plan 2000-2006. J. Kehoe is funded by the Teagasc Walsh Fellowship schem

Characterization of new membrane materials by means of fouling experiments Adsorption of bsa on polyetherimide-polyvinylpyrrolidone membranes

The hydrophilicity of polyetherimide-polyvinylpyrrolidone (PEI-PVP) microfiltration membranes can be adjusted by means of a suitable post-treatment. The influence of the nature of the membrane surface on fouling properties was studied using permeation experiments before and after exposure to a protein (BSA) solution and adsorption experiments with 14C labelled BSA. A correlation between the permeation experiments and the radiolabelled BSA adsorption experiments was found. The PVP in the membrane matrix prevents BSA adsorption taking place to a large extent and it appeared that heat-treated PEI-PVP membranes showed the same nonfouling behaviour as, for example, cellulose acetate membranes

Determination of the Optimal Conditions for Bovine Serum Albumin Surface Enhanced Raman Scattering on Silver Colloids

Bovine serum albumin (BSA) was analyzed using surface enhanced Raman scattering (SERS) to find the optimal conditions to observe BSA with SERS. Colloidal silver, Na2SO4, and BSA were mixed together at varying pHs and concentrations to obtain multiple spectra.The most favorable conditions using SERS for BSA were 500ug/mL and pH 4.The spectrum under those conditions showed the most intense and discernible peaks and the alpha helical secondary structure was very distinct at 1297 cm-1. SERS can be used for label free detection of proteins, thus finding the best conditions to obtain spectra using this technique may be very beneficial to proteomic research

Determination of the Optimal Conditions for Bovine Serum Albumin Using Surface Enhanced Raman Scattering on Silver Colloids and Nanoparticle Films

Bovine serum albumin (BSA) was analyzed using surface enhanced Raman scattering (SERS) to find the optimal conditions to observe BSA with SERS on colloids and nanoparticles films. The optimal conditions were determined by varying the concentrations of colloidal silver, Na2SO4, and BSA. The most favorable conditions using SERS for BSA were 500ug/mL, pH 4, and 0.1 M Na2SO4. Under these conditions, peaks due to an alpha helical secondary structure by the amide 3 vibrations at 1297 cm-1 were most distinct. The peaks appearing in the spectrum are the parts of the BSA molecule which are interacting with the silver colloids. Colloidal metal films were made and soaked in varying pH of 500 µg/mL BSA solutions, the spectrum at pH 4 showed tyrosine peaks at 1566 and 1339 cm-1 and tryptophan peaks at 1158 and 1038 cm-1. The peaks on colloidal films only appear when the amino acid residue is perpendicular to the colloidal surface; therefore, one could deduce that the tyrosine molecules at 1566 and 1339 cm-1 and tryptophan molecules at 1158 and 1038 cm-1 are perpendicular to the film. SERS can be used for label-free detection of proteins, thus finding the best conditions to obtain spectra using this technique may be very beneficial to proteomic research

Buffalo Sewer Authority

The Buffalo Sewer Authority is a public benefit corporation created by the New York State legislature in 1935 to clean wastewater before it is released into the environment. The BSA also maintains the storm drains for the City of Buffalo. The BSA serves the residents and businesses of the Buffalo area as well as some neighboring communities. Currently, around 98,000 Buffalo residents and nearly 400 businesses in the City of Buffalo are served by the BSA

Improved linkage analysis of Quantitative Trait Loci using bulk segregants unveils a novel determinant of high ethanol tolerance in yeast

Background: Bulk segregant analysis (BSA) coupled to high throughput sequencing is a powerful method to map genomic regions related with phenotypes of interest. It relies on crossing two parents, one inferior and one superior for a trait of interest. Segregants displaying the trait of the superior parent are pooled, the DNA extracted and sequenced. Genomic regions linked to the trait of interest are identified by searching the pool for overrepresented alleles that normally originate from the superior parent. BSA data analysis is non-trivial due to sequencing, alignment and screening errors. Results: To increase the power of the BSA technology and obtain a better distinction between spuriously and truly linked regions, we developed EXPLoRA (EXtraction of over-rePresented aLleles in BSA), an algorithm for BSA data analysis that explicitly models the dependency between neighboring marker sites by exploiting the properties of linkage disequilibrium through a Hidden Markov Model (HMM). Reanalyzing a BSA dataset for high ethanol tolerance in yeast allowed reliably identifying QTLs linked to this phenotype that could not be identified with statistical significance in the original study. Experimental validation of one of the least pronounced linked regions, by identifying its causative gene VPS70, confirmed the potential of our method. Conclusions: EXPLoRA has a performance at least as good as the state-of-the-art and it is robust even at low signal to noise ratio's i.e. when the true linkage signal is diluted by sampling, screening errors or when few segregants are available

Filtration characteristics of hollow fiber microfiltration membranes used in a specific double membrane bioreactor

The performance of the microfiltration in a specifically designed membrane bioreactor operating under various transmembrane pressures with periodic backwashing was investigated for model media. These media were representative of some usual components of a fermentation medium: BSA solution (2 g L−1), yeast suspension (8 g L−1, dry mass) and a mixture of BSA/yeast (2 g L−1/8 g L−1). In this system, the separation was provided by a 0.1 μm polysulfone hollow fiber membrane. The net permeate fluxes observed for yeast/BSA mixture were proportional to the transmembrane pressure applied (ΔP) but were less than those obtained with water osmosis, showing that, in spite of the periodic backwash, a small amount of irreversible fouling remained. This fouling can be assumed to be due to internal fouling by protein and/or external fouling by a residual yeast cake. Moreover, the net permeate flux obtained with the yeast/BSA mixture was higher than that obtained with the BSA alone, showing that a thin yeast cake probably acted as a primary filtration layer that could protect the polysulfone membrane against protein fouling. These experiments enable operating recommendations to be made for the use of this specific bioreactor concerning the transmembrane pressure value and the possible addition of inert particles

A pH-sensitive stearoyl-PEG-poly(methacryloyl sulfadimethoxine)-decorated liposome system for protein delivery: an application for bladder cancer treatment

Stealth pH-responsive liposomes for the delivery of therapeutic proteins to the bladder epithelium were prepared using methoxy-poly(ethylene glycol)5kDa-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (mPEG5kDa-DSPE) and stearoyl-poly(ethylene glycol)-poly(methacryloyl sulfadimethoxine) copolymer (stearoyl-PEG-polySDM), which possesses an apparent pKa of 7.2. Liposomes of 0.2:0.6:100, 0.5:1.5:100 and 1:3:100 mPEG5kDa-DSPE/stearoyl-PEG-polySDM/(soybean phosphatidylcholine + cholesterol) molar ratios were loaded with bovine serum albumin (BSA) as a protein model. The loading capacity was 1.3% w/w BSA/lipid. At pH 7.4, all liposome formulations displayed a negative zeta-potential and were stable for several days. By pH decrease or addition to mouse urine, the zeta potential strongly decreased, and the liposomes underwent a rapid size increase and aggregation. Photon correlation spectroscopy (PCS) and transmission electron microscopy (TEM) analyses showed that the extent of the aggregation depended on the stearoyl-PEG-polySDM/lipid molar ratio. Cytofluorimetric analysis and confocal microscopy showed that at pH 6.5, the incubation of MB49 mouse bladder cancer cells and macrophages with fluorescein isothiocyanate-labelled-BSA (FITC-BSA) loaded and N-(Lissamine Rhodamine B sulfonyl)-1, 2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine triethylammonium salt (rhodamine-DHPE) labelled 1:3:100 mPEG5kDa-DSPE/stearoyl-PEG-polySDM/lipid molar ratio liposomes resulted in a time-dependent liposome association with the cells. At pH 7.4, the association of BSA-loaded liposomes with the MB49 cells and macrophages was remarkably lower than at pH 6.5. Confocal images of bladder sections revealed that 2 h after the instillation, liposomes at pH 7.4 and control non-responsive liposomes at pH 7.4 or 6.5 did not associate nor delivered FITC-BSA to the bladder epithelium. On the contrary, the pH-responsive liposome formulation set at pH 6.5 and soon administered to mice by bladder instillation showed that, 2 h after administration, the pH-responsive liposomes efficiently delivered the loaded FITC-BSA to the bladder epitheliu

Will anyone vote? Prospects for turnout in the general election

One of the most notable features of the last two general elections was the low level of turnout. Before 2001 turnout at general elections was always at least 70% (and often far higher). But in 2001 it fell to just 59% and, at 61%, the figure in 2005 was little better. Over 17 million people eligible to vote that year chose not to do so, seven million more than voted for the winning Labour party. Britain found itself almost at the bottom of the turnout league among established European democracies. The failure of large sections of the public to go to the polls has led to considerable concern about the health of Britain’s democracy and stimulated many a suggestion as to how the country’s politicians might be able to reconnect with the electorate