Understanding the inuence of nanoparticles on the formation of protein amyloid brillation is cru-cial to extend their application in related biological diagnosis and nanomedicines.In this work,Ra-man spectroscopy was ...Understanding the inuence of nanoparticles on the formation of protein amyloid brillation is cru-cial to extend their application in related biological diagnosis and nanomedicines.In this work,Ra-man spectroscopy was used to probe the amyloid brillation of hen egg-white lysozyme in the pres-ence of silver nanoparticles(Ag-NPs)at di erent concentrations,combined with atomic force mi-croscopy and thioavin T(ThT)uorescence assays.Four representative Raman indicators were utilized to monitor transformation of the protein tertiary and secondary structures at the molecular level:the Trp doublet bands at 1340 and 1360 cm^(-1),the disul de stretching vibrational peak at 507 cm^(-1),the N-Cα-C stretching vibration at 933 cm^(-1),and the amide I band.All experimental results con rmed the concentration-dependent inuence of AgNPs on the hen egg-white lysozyme amyloid brillation kinetics.In the presence of AgNPs at low concentration(17μg/mL),electrostatic interaction of the nanoparticles stabilizes disul de bonds,and protects the Trp residues from exposure to hydrophilic environment,thus leading to formation of amorphous aggregates rather than brils.However,with the action of AgNPs at high concentration(1700μg/mL),the native disul de bonds of hen egg-white lysozyme are broken to form Ag-S bonds owing to the competition of electrostatic interaction from a great deal of nanoparticles.As for providing functional surfaces for protein to interact with,AgNPs play a bridge role in direct transformation from-helices to organized-sheets.The present investigation sheds light on the controversial e ects of AgNPs on the kinetics of hen egg-white lysozyme amyloid brillation.展开更多
为提高蛋清蛋白凝胶性,本研究以蛋清蛋白(Egg white protein,EWP)为研究对象,通过分析质构、持水率、分子作用力、傅里叶红外色谱、圆二色谱、粒径、电位、巯基含量、表面疏水力及扫描电镜的变化,探究超高压处理(Ultra-high pressure,U...为提高蛋清蛋白凝胶性,本研究以蛋清蛋白(Egg white protein,EWP)为研究对象,通过分析质构、持水率、分子作用力、傅里叶红外色谱、圆二色谱、粒径、电位、巯基含量、表面疏水力及扫描电镜的变化,探究超高压处理(Ultra-high pressure,UHP)、谷氨酰胺转胺酶处理(Transglutaminase,TG)及超高压协同TG酶处理(Ultra-high pressure synergistic Transglutaminase,UTG)的蛋清蛋白热诱导凝胶机理及结构的变化。结果表明:UHP-EWP、TG-EWP、UTG-EWP的硬度、弹性和持水性均有所提高,疏水相互作用力是维持凝胶的主要作用力;UHP-EWP、TG-EWP、UTG-EWP的α-螺旋含量均不同程度下降、β-折叠含量均上升;UHP-EWP的平均粒径值下降、电位绝对值下降,TG-EWP、UTG-EWP的变化与之相反;UHP-EWP、TG-EWP、UTG-EWP的游离巯基含量上升、总巯基含量下降、表面疏水性升高;凝胶结构更加致密光滑,平整度提高。本研究为蛋清蛋白热诱导凝胶改性提供了理论基础及研究思路。展开更多
本实验以蛋清蛋白微粒(egg white protein microparticles,EWPM)为代脂基质,研究EWPM/黄原胶(xanthan gum,XG)共混质量分数、pH值、氯化钠含量、糖含量、均质条件等对超低脂沙拉酱产品流变学特性的影响规律。研究发现,XG对EWPM基超低脂...本实验以蛋清蛋白微粒(egg white protein microparticles,EWPM)为代脂基质,研究EWPM/黄原胶(xanthan gum,XG)共混质量分数、pH值、氯化钠含量、糖含量、均质条件等对超低脂沙拉酱产品流变学特性的影响规律。研究发现,XG对EWPM基超低脂沙拉酱具有显著的增稠效果,然而含量过高会导致低脂沙拉酱恢复性下降。基础调味组分对超低脂沙拉酱品质的影响存在较大差异,pH值和盐对体系黏度影响较大,糖添加量对黏度的影响相对较小。适度的胶体磨均质转速和均质时间可以减小产品的粒径,增加其内部均一性,显著提高产品的整体黏度。本实验以沙拉酱流变学行为调控为重点,为低脂沙拉酱的研究提供一个更加明确的工艺优化方向。展开更多
建立高脂小鼠模型,以辛伐他汀和市售醋蛋液水解物作为药品和食品阳性对照组,综合评价肽酶433P酶解蛋清获得蛋清水解物(egg white hydrolysate,EWH)的体内降血脂效果。结果表明,EWH可以显著抑制小鼠体质量增长,调节血脂异常,减轻肝脏脂...建立高脂小鼠模型,以辛伐他汀和市售醋蛋液水解物作为药品和食品阳性对照组,综合评价肽酶433P酶解蛋清获得蛋清水解物(egg white hydrolysate,EWH)的体内降血脂效果。结果表明,EWH可以显著抑制小鼠体质量增长,调节血脂异常,减轻肝脏脂质异常与炎症损伤,抑制肝脏脂肪细胞的积累、变性和脂肪组织甘油三酯的堆积。之后研究了胃肠消化对EWH血脂调节活性影响,结果发现,经胃肠消化后EWH的胆固醇胶束溶解度抑制率下降,但胆汁酸结合率和胰脂肪酶抑制率基本无明显变化。最后通过质谱鉴定及分子对接,得到了3个潜在的胰脂肪酶抑制活性肽,其氨基酸序列分别为LWVPSVY、YPILPEYLQ和WNIPIGTL,上述肽段可以通过氢键、疏水相互作用和静电相互作用等作用力与受体蛋白1ETH相互作用,从而发挥降血脂作用。展开更多
以蛋清蛋白(egg white protein,EWP)和黄油为原料,按照不同比例(1∶1、1∶2、1∶3、1∶4、1∶5,质量比)进行复配,经热诱导制备EWP/黄油复合凝胶,并分析其质构特性、持水性、乳化性、乳化稳定性、流变特性、扫描电镜及激光共聚焦的变化...以蛋清蛋白(egg white protein,EWP)和黄油为原料,按照不同比例(1∶1、1∶2、1∶3、1∶4、1∶5,质量比)进行复配,经热诱导制备EWP/黄油复合凝胶,并分析其质构特性、持水性、乳化性、乳化稳定性、流变特性、扫描电镜及激光共聚焦的变化规律。结果表明,质构特性中的硬度及胶黏性随着黄油添加量的增加而增加,而弹性及内聚性则在EWP/黄油为1∶3(质量比)时达到最高(P<0.05);复合凝胶的持水性随着黄油添加量的增加而增加,在1∶4及1∶5(质量比)时差异不显著(P>0.05);当二者质量比为1∶3时,乳化性及乳化稳定性显著高于其他组(P<0.05);流变结果显示当二者质量比为1∶3时,复合凝胶具有较大的储能模量(G′)及较低的损耗因子tanδ,说明此比例下的黄油提高了复合凝胶的黏弹性;扫描电镜结果表明二者质量比为1∶3时促使复合凝胶形成了更致密紧凑的网络结构,而未添加黄油的蛋清蛋白凝胶则呈现孔洞较大且大小不均匀的结构特征;激光共聚焦结果显示EWP/黄油1∶3(质量比)时网络结构相对均匀,且无析水无孔隙。综上所述,EWP和黄油的质量比为1∶3时可以改善蛋清蛋白的乳化特性及凝胶特性,为后续替代食品中部分乳化盐,以降低食品中钠含量,制备相关低钠食品提供了数据支撑。展开更多
基金This work was supported by the National Natu-ral Science Foundation of China(No.22073088 and No.21873089).
文摘Understanding the inuence of nanoparticles on the formation of protein amyloid brillation is cru-cial to extend their application in related biological diagnosis and nanomedicines.In this work,Ra-man spectroscopy was used to probe the amyloid brillation of hen egg-white lysozyme in the pres-ence of silver nanoparticles(Ag-NPs)at di erent concentrations,combined with atomic force mi-croscopy and thioavin T(ThT)uorescence assays.Four representative Raman indicators were utilized to monitor transformation of the protein tertiary and secondary structures at the molecular level:the Trp doublet bands at 1340 and 1360 cm^(-1),the disul de stretching vibrational peak at 507 cm^(-1),the N-Cα-C stretching vibration at 933 cm^(-1),and the amide I band.All experimental results con rmed the concentration-dependent inuence of AgNPs on the hen egg-white lysozyme amyloid brillation kinetics.In the presence of AgNPs at low concentration(17μg/mL),electrostatic interaction of the nanoparticles stabilizes disul de bonds,and protects the Trp residues from exposure to hydrophilic environment,thus leading to formation of amorphous aggregates rather than brils.However,with the action of AgNPs at high concentration(1700μg/mL),the native disul de bonds of hen egg-white lysozyme are broken to form Ag-S bonds owing to the competition of electrostatic interaction from a great deal of nanoparticles.As for providing functional surfaces for protein to interact with,AgNPs play a bridge role in direct transformation from-helices to organized-sheets.The present investigation sheds light on the controversial e ects of AgNPs on the kinetics of hen egg-white lysozyme amyloid brillation.
文摘本实验以蛋清蛋白微粒(egg white protein microparticles,EWPM)为代脂基质,研究EWPM/黄原胶(xanthan gum,XG)共混质量分数、pH值、氯化钠含量、糖含量、均质条件等对超低脂沙拉酱产品流变学特性的影响规律。研究发现,XG对EWPM基超低脂沙拉酱具有显著的增稠效果,然而含量过高会导致低脂沙拉酱恢复性下降。基础调味组分对超低脂沙拉酱品质的影响存在较大差异,pH值和盐对体系黏度影响较大,糖添加量对黏度的影响相对较小。适度的胶体磨均质转速和均质时间可以减小产品的粒径,增加其内部均一性,显著提高产品的整体黏度。本实验以沙拉酱流变学行为调控为重点,为低脂沙拉酱的研究提供一个更加明确的工艺优化方向。
文摘建立高脂小鼠模型,以辛伐他汀和市售醋蛋液水解物作为药品和食品阳性对照组,综合评价肽酶433P酶解蛋清获得蛋清水解物(egg white hydrolysate,EWH)的体内降血脂效果。结果表明,EWH可以显著抑制小鼠体质量增长,调节血脂异常,减轻肝脏脂质异常与炎症损伤,抑制肝脏脂肪细胞的积累、变性和脂肪组织甘油三酯的堆积。之后研究了胃肠消化对EWH血脂调节活性影响,结果发现,经胃肠消化后EWH的胆固醇胶束溶解度抑制率下降,但胆汁酸结合率和胰脂肪酶抑制率基本无明显变化。最后通过质谱鉴定及分子对接,得到了3个潜在的胰脂肪酶抑制活性肽,其氨基酸序列分别为LWVPSVY、YPILPEYLQ和WNIPIGTL,上述肽段可以通过氢键、疏水相互作用和静电相互作用等作用力与受体蛋白1ETH相互作用,从而发挥降血脂作用。
文摘以蛋清蛋白(egg white protein,EWP)和黄油为原料,按照不同比例(1∶1、1∶2、1∶3、1∶4、1∶5,质量比)进行复配,经热诱导制备EWP/黄油复合凝胶,并分析其质构特性、持水性、乳化性、乳化稳定性、流变特性、扫描电镜及激光共聚焦的变化规律。结果表明,质构特性中的硬度及胶黏性随着黄油添加量的增加而增加,而弹性及内聚性则在EWP/黄油为1∶3(质量比)时达到最高(P<0.05);复合凝胶的持水性随着黄油添加量的增加而增加,在1∶4及1∶5(质量比)时差异不显著(P>0.05);当二者质量比为1∶3时,乳化性及乳化稳定性显著高于其他组(P<0.05);流变结果显示当二者质量比为1∶3时,复合凝胶具有较大的储能模量(G′)及较低的损耗因子tanδ,说明此比例下的黄油提高了复合凝胶的黏弹性;扫描电镜结果表明二者质量比为1∶3时促使复合凝胶形成了更致密紧凑的网络结构,而未添加黄油的蛋清蛋白凝胶则呈现孔洞较大且大小不均匀的结构特征;激光共聚焦结果显示EWP/黄油1∶3(质量比)时网络结构相对均匀,且无析水无孔隙。综上所述,EWP和黄油的质量比为1∶3时可以改善蛋清蛋白的乳化特性及凝胶特性,为后续替代食品中部分乳化盐,以降低食品中钠含量,制备相关低钠食品提供了数据支撑。