CdS-BSA-CTMAB荧光光度法测定食品中蛋白质的含量

实验以六偏磷酸钠为稳定剂,巯基乙酸为修饰剂水相合成了荧光试剂CdS量子点。结果表明,牛血清白蛋白(BSA)可使CdS的荧光峰增强,而阳离子表面活性剂十六烷基三甲基溴化铵(CTMAB)的加入,可使体系荧光峰显著增敏,并且荧光强度与BSA浓度在8.0×10-5～1.3mg/mL范围内呈良好的线性关系,回归方程为F=1486.94793+937.70328C,相关系数R=0.9957,检出限为7.4×10-5mg/mL。方法已用于食品中蛋白质的测定,与考马斯亮蓝法对照,结果满意。     

柿子单宁对牛血清白蛋白的荧光猝灭作用研究

采用荧光光谱和紫外吸收光谱法研究了柿子单宁对牛血清白蛋白(BSA)的荧光猝灭作用.结果表明,柿子单宁可以有规律地使BSA内源荧光猝灭,其猝灭机理可认为是柿子单宁与BSA形成复合物的静态猝灭,并获得了不同温度下柿子单宁与BSA作用的结合常数和热力学参数.根据所得结果可推断柿子单宁与BSA的作用力为疏水作用力和静电作用力,同时由Forster非辐射能量转移理论计算得出了柿子单宁与BSA结合位置的距离.     

聚醚砜微孔膜对牛血清蛋白吸附性能的研究

主要研究了聚醚砜(PES)微孔膜对牛血清蛋白(BSA)的吸附性能.用BCA(bicinchoninincacid)法测定了膜孔中吸附的蛋白量.运用扫描电子显微镜和电子单纱强力仪分别对PES膜结构和力学性能进行表征.随着聚乙二醇20000(PEG20000)与聚乙二醇400(PEG400)比值的增大牛血清蛋白的吸附量先减小后增大,水通量先增大后减小,截留率始终维持在97.93%～99.39%之间,结果表明,PEG20000与PEG400比例为2时PES膜的亲水性最好,对BSA的吸附量最小.     

Electroseparation of an antibacterial peptide fraction from snow crab by-products hydrolysate by electrodialysis with ultrafiltration membranes

Recently, a snow crab by-products hydrolysate has demonstrated antibacterial properties due to a peptide with a molecular weight of about 800 Da, but only at high concentration. Consequently, peptide hydrolysate has been fractionated to obtain peptides in a more purified form. The aim of this work was to separate a snow crab by-products hydrolysate by electrodialysis with ultrafiltration membranes (EDUF). EDUF, which allows separation of molecules according to their charges and molecular weights, was used to recover and concentrate the active antibacterial fraction. Two different ultrafiltration membranes (20 and 50 kDa) and two electrical field strengths (2 and 14 V/cm) were used as separation parameters. After EDUF separation, the 300-600 Da peptide molecular weight range was the most recovered with an abundance of 94%. Moreover, fractionation at 14 V/cm with ultrafiltration membranes of 50 kDa allowed the recovery of an anionic fraction which showed antibacterial properties on Escherichia coli ATCC 25922 and Listeria innocua HPB 13.     

Proteomics in the Diagnosis of Endometriosis: Opportunities and Challenges

The non‐surgical diagnosis of endometriosis is still challenging for the clinician. Ultrasonography and magnetic resonance imaging can be used to diagnose ovarian endometriotic cysts and deep infiltrating endometriosis; but their performance is poor in the diagnosis of initial stages of endometriosis. CA‐125 and other serum markers (such as CA 19‐9, serum protein PP14, interleukins, and angiogenetic factors) have been measured in women with endometriosis but they are not reliable for the diagnosis of the disease. Although several studies used proteomics technologies to identify plasmatic markers of endometriosis, the non‐invasive diagnosis of endometriosis is far from being achieved. In this issue, Manousopoulou et al. compare the integrated quantitative proteomic profile of eutopic endometrium and serum of women with endometriosis and controls. 1214 proteins are differentially expressed in the eutopic endometrium and 404 proteins in the serum of the two study groups. 21 proteins are aberrantly expressed in both eutopic endometrium and serum of women with endometriosis. More work is needed to assess if the differentially expressed proteins identified in this study can be used as clinical markers of endometriosis.     

Investigation of the Binding Behavior between the S-heterocyclic Aromatic Palladium(II) Complex and Human Serum Albumin: Spectroscopic Approach

The interaction of 1, 10-phenanthroline octhyldithiocarbamato palladium(II) nitrate ([Pd(Oct-dtc)(phen)]NO₃) with human serum albumin (HSA) has been investigated by various spectroscopic techniques under physiological conditions. Here, HSA was titrated with the Pd(II) complex, followed by UV–Vis absorption spectroscopy to estimate a binding constant (K_b) and other thermodynamic parameters. The results indicate that the Pd (II) complex has a high affinity for bind HSA. Thermodynamic analysis showed that the enthalpy (ΔH°) and entropy changes (ΔS°) are positive and Gibbs free energy change (ΔG°) is negative which indicated that hydrophobic interactions played the predominant role in the binding process. Fluorescence spectroscopy were used to show the mechanism and binding parameters of this interaction. Utilizing the Stern–Volmer equation, the Pd(II) complex quenched the intrinsic fluorescence of HSA via a static quenching procedure. The specific binding distances between the tryptophan (donor) proteins and Pd(II) complex (acceptor) were estimated by Forster resonance energy transfer. The CD results also showed the conformational changes on serum albumin upon binding with the Pd(II) complex.     

Efficient adsorptive extraction materials by surface protein-imprinted polymer over silica gel for selective recognition/separation of human serum albumin from urine

In this study, we report the development of adsorptive extraction materials by surface protein-imprinted polymers (MIPs) over silica gel for selective recognition/separation of human serum albumin (HSA) from urine. The HSA-imprinted polymers prepared on silica particle had at interface between the silica gel and different MIPs greatly produced enrichment for the binding of protein from the urine. The solid-phase extraction of the optimized polymer layer was prepared by copolymerization of methacrylic acid (MAA), acrylamide (AAm), and dimethylaminoethylmethacrylate (DMAEMA) and a crosslinker methylenebisacrylamide (MBA) at the mole ratio of 1:158:88 (T:M:C) and showed moderate affinity (<10⁴ order M⁻¹) toward target protein HSA and selectivity. Four analogues, egg white albumin (EWA), bovine serum albumin (BSA), lysozyme (Lyz), and creatinine (Cre) were selected to study the binding efficiency of MIPs in single and binary protein solutions. We studied the influence on recognition ability for HSA and found that prepolymer mixture and matrix flexibility of the optimized thin polymer layer (35 ± 10 nm) on the submicrosilica particles. The high-binding affinity (Q_MIP, 86.7 mg g⁻¹) and fast kinetics (180 min) were observed for this synthesized HSA-MIP when compared with other reported HSA-MIPs in surface imprinting (5.9 and 11.3 mg g⁻¹) and epitope surface imprinting (46.6 mg g⁻¹) methods. We demonstrated the application in real and synthetic urine samples that the approach allowed the efficient adsorption of HSA in real urine (129.48 mg g⁻¹) is almost double to the binding of HSA in synthetic urine (67.84 mg g⁻¹). Apart from this, only minor interference of Cre (2.74 mg g⁻¹) was observed, eventhough Cre is the final metabolite in urine. These adsorptive submicrosilica materials have potential in the pharmaceutical industry and clinical analysis applications. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 46894.     

Blood parameters of young calves at abattoirs are related to distance transported and farm of origin

Nonreplacement dairy calves, or bobby calves, are fasted and transported to abattoirs from as young as 5 d of age in Australia. The aims of this cross-sectional observational study were (1) to assess the welfare status, as measured by blood parameters, of bobby calves in the commercial supply chain after transport and lairage, and (2) to assess whether distance and duration of transport are risk factors for poor bobby calf welfare, as measured by blood parameters. We hypothesized that bobby calves transported greater distances would be more likely to show evidence of compromised welfare, as measured by blood indicators of hydration, energy status, and muscle fatigue or damage. We also hypothesized that there would be a large amount of variability in indicators of energy status between calves from different farms. We analyzed blood samples collected at slaughter over a spring and an autumn calving period from 4,484 Australian bobby calves aged approximately 5 to 14 d old from 3 different states, after transport, fasting, and lairage. Packed cell volume (PCV), plasma glucose, and serum urea, total protein, β-hydroxybutyrate (BHB), and creatine kinase (CK) were measured. Radio frequency identification ear tag data were used to estimate the distance that the calves were transported and to identify the farm of origin. Data were analyzed using linear mixed models, except for BHB, which was analyzed using a Goodman-Kruskal gamma test due to left censoring of the data. Twelve percent of calves showed evidence of anemia (PCV less than 0.23 L/L), and 11% had urea concentrations consistent with dehydration (urea more than 7.7 mmol/L). Thirty-six percent of calves had CK activity above normal resting values, and 1% of calves had CK >2,000 U/L, indicating muscle fatigue or damage. Distance transported had significant effects on all blood variables except urea and BHB. With increasing distance transported, calves were more likely to show evidence of a negative energy balance (low plasma glucose) or dehydration (high PCV or total protein). The estimated effect of distance overall was small, but for calves transported more than 500 km, plasma glucose concentration declined more per kilometer. The calves' farm of origin accounted for a reasonable amount of the random variation between calves for plasma glucose (20%). Our results suggest that longer transport distances may increase the risk of poor calf welfare (dehydration, negative energy balance) after transport, and on-farm calf management (e.g., nutrition, timing of feeding before transport) may affect transported calves' energy status; improving this area could result in better energy availability during fasting.     

Interactions of different polyphenols with bovine serum albumin using fluorescence quenching and molecular docking

Polyphenols are responsible for the major organoleptic characteristics of plant-derived foods and beverages. Here, we investigated the binding of several polyphenols to bovine serum albumin (BSA) at pH 7.5 and 25 °C: catechins [(−)-epigallocatechin-3-gallate, (−)-epigallocatechin, (−)-epicatechin-3-gallate], flavones (kaempferol, kaempferol-3-glucoside, quercetin, naringenin) and hydroxycinnamic acids (rosmarinic acid, caffeic acid, p-coumaric acid). Fluorescence emission spectrometry and molecular docking were applied to compare experimentally determined binding parameters with molecular modelling. Among these polyphenols, (−)-epicatechin-3-gallate showed the highest Stern–Volmer modified quenching constant, followed by (−)-epigallocatechin-3-gallate. Similarly, (−)-epicatechin-3-gallate had the highest effect on the Circular Dichroic spectrum of BSA, while the changes induced by other polyphenols were negligible. Molecular docking predicted high binding energies for (−)-epicatechin-3-gallate and (−)-epigallocatechin-3-gallate for the binding site on BSA near Trp213. Our data reveal that the polyphenol structures significantly affect the binding process: the binding affinity generally decreases with glycosylation and reduced numbers of hydroxyl groups on the second aromatic ring.