pH对洛美沙星与BSA之间结合常数的影响

[目的]研究洛美沙星与血清白蛋白之间的作用,了解药物分子在体内的运输和分布情况。[方法]采用毛细管区带电泳法,通过测定在不同pH值、不同牛血清白蛋白浓度的缓冲溶液条件下药物迁移时间的变化,计算出洛美沙星与牛血清白蛋白相互作用的结合常数（Kb）。[结果]pH分别为6.8,7.4和8.0时,洛美沙星与BSA之间的Kb分别为10.185×1045、.319×1045、.356×104 L/mol。[结论]环境pH对洛美沙星与BSA之间的相互作用有强烈影响,当pH处在6.8～7.4之间时,Kb对pH尤为敏感。     

花生清蛋白的分离纯化及抗氧化、DNA酶活性研究

为了解花生清蛋白的生物学活性,本试验采用硫酸铵沉淀法初步分离花生种子中的花生清蛋白,并用DEAE-52柱纯化得到花生清蛋白,分析其体外抗氧化性及DNA酶活性。结果表明,65%硫酸铵分离花生清蛋白效果最好;纯化后的花生清蛋白由3个亚基组成,其相对分子质量分别为14.5、15.5、17.2 kDa。花生清蛋白体外抗氧化活性较高,对DPPH自由基和羟基自由基的清除率与花生清蛋白浓度呈正相关,清蛋白浓度从0.02 mg·mL^-1升至0.10 mg·mL^-1时,DPPH自由基清除率从26.3%增加到45.0%,羟基自由基清除率从10.8%增加到93.5%。DNA酶活性也与花生清蛋白浓度呈正相关,清蛋白浓度从0.1 mg·mL^-1提高到0.6 mg·mL^-1,清蛋白和质粒混合液的电泳条带逐渐变暗;当花生清蛋白浓度为0.8 mg·mL^-1时,电泳条带消失,质粒被完全降解;Ca²⁺、Mg²⁺、K⁺、Na⁺4种金属离子都有增强清蛋白DNA酶活性的作用,增强能力由大到小依次是Mg²⁺>Ca²⁺>Na⁺>K⁺。本研究结果为花生清蛋白功能性质研究及开发利用提供了一定的理论基础。     

几种添加物对蛋清蛋白热诱导不可逆凝胶性质的影响

以蛋清蛋白为原料,制备热诱导制取不可逆性凝胶,在单因素试验基础上,采用三因素二次正交旋转组合设计研究了三聚磷酸钠(X1)、瓜尔胶(X2)、酪蛋白酸钠(X3)对蛋清蛋白热诱导不可逆性凝胶性质的影响,建立了二次回归模型。结果表明,X12,X22在0.01水平上差异极显著,X32在0.05水平上差异显著;最佳配比为三聚磷酸钠质量分数0.100%,瓜尔胶质量分数0.100%,酪蛋白酸钠质量分数0.250%,此时蛋清蛋白凝胶强度预测值为550 g/cm2,实际值为542 g/cm2,二次正交旋转组合设计法可信度强,具有很好的预测意义。     

Antioxidant,anti-amylase and anti-glycation potential of brans of some Sri Lankan traditional and improved rice (Oryza sativa L.) varieties

Brans of 23 traditional and 12 improved (both red and white) rice varieties in Sri Lanka were screened for anti-amylase and anti-glycation activities in vitro. Varieties which showed the highest inhibitory activities at screening were further investigated for anti-glucosidase and glycation reversing as anti-diabetic properties. The same varieties were studied for selected antioxidant properties. Significantly high anti-amylase and anti-glycation activities were observed for bran extracts of red varieties compared to white varieties at screening. Traditional red rice varieties, Masuran, Sudu Heeneti, Dik Wee and Goda Heeneti, exhibited significant and dose dependent anti-amylase, anti-glycation and glycation reversing activities. These varieties also showed marked antioxidant properties. It is concluded that brans of Sri Lankan traditional red rice varieties Masuran, Sudu Heeneti, Dik Wee and Goda Heeneti may be potential food supplements for diabetes.     

Binding of phenthoate to bovine serum albumin and reduced inhibition on acetylcholinesterase

Organophosphorus pesticides (OPs) are of environmental significance due to their high toxicity to animals. Binding to plasma proteins may effective influence the toxicological properties of xenobiotics. In an attempt to evaluate the affinity of phenthoate (PTA) to bovine serum albumin (BSA) and inhibitory ability of bound PTA to acetylcholinesterase (AChE), we investigated the interactions between phenthoate (PTA) and bovine serum albumin (BSA) using tryptophan fluorescence quenching and subsequent inhibition on AChE activity by PTA. The results showed that PTA caused the fluorescence quenching of BSA because of the formation of a PTA-BSA complex. Quenching constants (K_sv), determined using the Sterns-Volmer equation to provide a measure of the binding affinity between PTA and BSA at 303, 306, 310 and 313 K were (3.4295 ± 0.0763) × 10⁻⁴, (3.2446 ± 0.0635) × 10⁻⁴, (3.0434 ± 0.0856) × 10⁻⁴ and (2.8262 ± 0.0569) × 10⁻⁴ M⁻¹, respectively. The thermodynamic parameters, ΔH and ΔS were −25.04 kJ mol⁻¹ and 168.94 J mol⁻¹ K⁻¹, respectively, which indicated that the electrostatic interactions played a major role in PTA-BSA association. The presence of BSA consistently reduced the inhibitory ability of PTA on AChE, with the relative activity being increased from 46.98 to 61.71% for the concentration range of BSA between 0 and 4.0 g L⁻¹.     

二苯-α-吡喃酮类链格孢霉毒素和人血清白蛋白的相互作用及机理探究

为探究链格孢酚单甲醚(AME)和链格孢酚(AOH)这两种二苯-α-吡喃酮类链格孢霉毒素与人血清白蛋白(HSA)之间的分子相互作用及机理,本研究模拟血液生理pH值,采用稳态荧光光谱、同步荧光光谱、3D荧光光谱以及圆二色光谱等方法研究两者之间的相互作用。结果表明,AME和AOH与HSA相互作用均发生了静态猝灭,具有较高亲和力,以氢键和范德华力(ΔH<0,ΔS<0)结合形成1:1复合物,互作过程中这两种类毒素破坏了人血清白蛋白中稳定二级结构的氢键网络,使蛋白二级结构展开,且AME和AOH使人血清白蛋白中的α-螺旋结构含量从48.93%分别减少至39.41%和44.01%,并使色氨酸残基极性降低、疏水性增加,但酪氨酸残基极性变化不大,即结合位点可能位于色氨酸残基所在的空腔(即结合位点I);AME对HSA的猝灭程度、猝灭常数(K_sv)及结合常数(K_a)均大于AOH,结合距离也更小(r_AME=2.56 nmAOH=2.60 nm)。本研究结果为进一步探究二苯-α-吡喃酮类链格孢霉毒素的毒代动力学和药代动力学,完善丰富其毒理学相关资料,以及进行风险评估和在食品中限量标准的制定提供了参考依据。     

诺氟沙星与牛血清白蛋白及卵清蛋白结合物的合成

采用碳二亚胺法制备诺氟沙星与牛血清白蛋白（BSA）的结合物，作为诺氟沙星酶联免疫用的免疫原。经紫外光谱法测定，每分子牛血清白蛋白连接的诺氟沙星分子数为16.1个。分别用碳二亚胺法和混合酸酐法制备诺氟沙星与卵清蛋白（OA）的结合物，作为诺氟沙星酶联免疫测定用的包被抗原。经紫外光谱法测定，每分子卵清蛋白连接的诺氟沙星分子数分别为5.8和1.9个。     

磺胺对甲氧嘧啶全抗原的制备研究

采用戊二醛法 ,将半抗原磺胺对甲氧嘧啶 (SMD)与载体牛血清白蛋白 (BSA)偶联 ,制得磺胺对甲氧嘧啶全抗原 SMD-BSA。其紫外扫描光谱和红外光谱表明偶联成功。通过改变半抗原 SMD和偶联剂戊二醛的用量 ,制得多种结合比的全抗原 ,这为进一步制备抗 SMD抗体提供了良好的免疫原