酪蛋白糖巨肽分离纯化方法的研究进展

酪蛋白糖巨肽(CGMP)主要是κ-酪蛋白经凝乳酶降解产生的一类含有糖链的多肽,具有许多的生理活性功能和独特的营养特性,可广泛地应用于保健食品和医药品。本文对酪蛋白糖巨肽的来源及结构作了简要概括,对其分离纯化方法及进展作出了详细论述。     

酪蛋白糖巨肽超滤分离的工艺优化研究

目的:研究超滤分离酪蛋白糖巨肽的最优工艺参数,以达到最佳超滤通量性能。方法:采用超滤方法分离乳清浓缩蛋白(WPC80)中的酪蛋白糖巨肽,研究料液pH值、操作压力和温度等对膜渗透通量的影响。利用SDS-PAGE凝胶电泳技术分析超滤截留液的相对分子量分布。结果:实验表明溶液pH7.0、操作压力0.3MPa、温度45℃时膜渗透通量最高,超滤酪蛋白糖巨肽的截留率达到97.54%,产物纯度达到92.45%,经SDS-PAGE凝胶电泳测定,酪蛋白糖巨肽的平均分子量68.86%为30.47ku,31.14%为19.22ku。结论:超滤技术是将酪蛋白糖巨肽与其他乳清蛋白分离的有效方法。     

乳源酪蛋白糖巨肽超滤分离的工艺优化研究

采用超滤方法分离酶解液中的酪蛋白糖巨肽,通过对料液pH值、操作压力和温度等对膜通量的影响,确定最优工艺参数,以达到最佳超滤通量性能,利用液相色谱法测定纯化后的酪蛋白糖巨肽分子量.试验结果表明:溶液pH9.0、操作压力0.3MPa、温度55℃时膜通量最高,超滤酪蛋白糖巨肽的截留率达到98.23%,产物纯度达到92.62%,酪蛋白糖巨肽的平均分子量为35682u.超滤方法分离酶解液中的酪蛋白糖巨肽是完全可行的.     

酪蛋白糖巨肽的生物学活性

酪蛋白糖巨肽(Caseino Glycomacropeptide,简称CGMP)主要是κ-酪蛋白经凝乳酶降解产生的一类含有糖链的多肽,具有许多的生理活性功能和独特的营养特性,可广泛地应用于保健食品和医药品.综述了CGMP的一些生理功能研究进展以及其生产制备工艺的现状,同时也介绍了它在食品和医药品中的应用前景.     

壳聚糖法分离纯化酪蛋白糖巨肽

探索壳聚糖法分离乳清粉中酪蛋白糖巨肽的工艺条件,通过单因素试验考察混合液pH值、壳聚糖添加量、吸附时间、氯化钠浓度和pH值对酪蛋白糖巨肽分离效果的影响。结果表明最佳分离工艺条件为:在混合液pH4.6时,采用质量浓度2g/L的壳聚糖吸附60min,再用2mol/L、pH9的氯化钠溶液进行洗脱。洗脱液经过超滤除盐、浓缩和冷冻干燥后可获得酪蛋白糖巨肽干粉。采用上述工艺进行放大实验,酪蛋白糖巨肽和唾液酸的回收率分别为83.3%和70.64%,每克酪蛋白糖巨肽含有10mg唾液酸。电泳检测结果显示制备的酪蛋白糖巨肽主要是分子质量约22ku和30ku的酪蛋白糖巨肽多聚体。     

酪蛋白糖巨肽的分离纯化研究

酪蛋白糖巨肽(CGMP)主要是-酪蛋白经凝乳酶降解产生的一类含有糖链的多肽,具有许多的生理活性功能和独特的营养特性,可广泛地应用于保健食品和医药品。对酪蛋白糖巨肽的适合于工业化生产的分离纯化条件进行更深层次的研究。     

乳清中酪蛋白糖巨肽的分离纯化

以酶凝干酪乳清为原料,通过沉淀、透析、超滤和离子交换层析等操作,分离提取酪蛋白糖巨肽,通过单因素分析和正交实验,优化出最佳提取工艺条件,同时采用SDS-聚丙烯酰胺凝胶电泳进行分析.结果表明:硫酸铵沉淀乳清中杂蛋白终饱和度为20%;超滤法纯化最佳工艺条件为温庹45℃,压力0.2 MPa,时间30 min;D201GF离子交换层析处理后制得的酪蛋白糖巨肽的纯度为83.97%;SDS-聚丙烯酰胺凝胶电泳结果显示,酪蛋白糖巨肽的相对分子量在15 ku左右.     

超滤法分离纯化酪蛋白糖巨肽的研究

酪蛋白糖巨肽(Casein glycoma cropeptide,简称CGMP)是蛋白酶酶解κ-酪蛋白得到的C末端亲水性糖肽,具有多种生理功能。研究了超滤法分离纯化CGMP的条件,得到超滤的最佳条件是在室温下,压差为0.02MPa,浓缩比为8。通过此方得到了比较纯的CGMP,蛋白回收率为1.77%,糖基化程度(唾液酸/蛋白质)为70.1μg/mg,并且此方法适用于工业化生产。     

酪蛋白糖巨肽分离条件的研究

探讨了利用了(NH4)2SO4分级沉淀、三氯乙(TCA)纯化酪蛋白的胃蛋白酶水解物中 CGMP的工艺.结果表明,经过20%和65%饱和度的硫酸铵沉淀胃蛋白酶水解物,质量分数为12%的TCA纯化得到的样品,其纯度为91.2%.整个工艺路线简便、快捷、成本较低,适合工业化生产.     

辣木凝乳酶水解酪蛋白位点及其酪蛋白磷酸肽、酪蛋白糖巨肽

基于十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,液相色谱-串联质谱法研究辣木凝乳酶对酪蛋白的酶切位点和水解特性,并分析该凝乳酶水解产生的酪蛋白糖巨肽和酪蛋白磷酸肽.结果表明,辣木凝乳酶酶切κ-酪蛋白的Arg 93-His 94位点导致凝乳,区别于其他已报道的凝乳酶,具有明显特异性.酶作用下κ-酪蛋白的米氏常数Km=0.49 m...     

酪蛋白糖巨肽(CGMP)生物活性的初步研究

酪蛋白糖巨肽(Casein Glycomacropeptide,CGMP)是一种来源于哺乳动物κ-酪蛋白中的具有64个氨基酸残基(106-109)的糖肽。它具有许多的生理活性功能和独特的营养特性。可以广泛的添加于保健食品和医药品。本文研究了用蛋白酶水解制备的酪蛋白糖巨肽的生物学活性,即抑制口腔致病菌-变形链球菌的生长、产酸和黏附的功能。     

超滤法纯化酪蛋白凝乳酶水解物中的酪蛋白糖巨肽

酪蛋白糖巨肽(CGMP)是蛋白酶酶解κ-酪蛋白得到的C末端亲水性糖肽,具有多种生理功能.研究了超滤法分离纯化CGMP的条件,得到超滤的最佳条件是在室温下,压差为0.02MPa,浓缩比为8.通过此方得到了比较纯的CGMP,蛋白回收率为1.77%,糖基化程度(唾液酸/蛋白质)为70.1 mg/g,并且此方法适用于工业化生产.     

凝乳酶水解酪蛋白生产酪蛋白糖巨肽工艺条件的研究

酪蛋白糖巨肽(casein glycomacropeptide,简称CGMP)是K-酪蛋白经酶解后生成的一类舍有糖链的多肽,具有许多功能特性.本文研究了凝乳酶水解酪蛋白生产CGMP的工艺条件,通过正交实验得到最佳工艺条件是底物浓度10mg/mL、酶的添加量为0.6%、酶解时间为2.5h.唾液酸回收率为80.7%,蛋白质回收率为3.03%,糖基化程度(唾液酸/蛋白质)为77.6μg/mg.     

Raman spectra of micellar casein powders prepared with wet blending of glycomacropeptide and micellar casein concentrate

Understanding the molecular properties and interactions of components within high-protein dairy powders is important in predicting rehydration performance. Raman spectroscopy was used to generate molecular information which provided insights into interactions between caseins in micellar casein concentrate (MCC) powder formulated with varying proportions of glycomacropeptide (GMP). The Raman bands showed intense peaks at 1446 to 1476 cm⁻¹ (due to CH₂ vibration) and 1653 to 1668 cm⁻¹ (due to C=O of Amide I and C=C bond) in MCC powder samples blended with GMP. Enhanced rehydration characteristics of MCC powder containing GMP matched with these differences in Raman peak intensity.     

酪蛋白糖巨肽对花生过敏原免疫反应性的影响

目的 探究酪蛋白糖巨肽与花生过敏原相互作用并降低其免疫反应性的潜力。方法 通过蛋白-蛋白分子对接技术探讨酪蛋白糖巨肽(casein glycomacropeptides, CGMP)与Ara h1、Ara h2是否有相互作用的潜力。进一步通过混合水浴加热制备CGMP与花生蛋白的混合溶液(mixed solution of casein glycomacropeptides and peanut proteins, MCGP),建立MCGP致敏、花生蛋白激发的BALB/c小鼠模型,研究MCGP对花生过敏反应的影响。最后使用圆二色谱法研究酪蛋白糖巨肽与Ara h1、Ara h2的相互作用力及对其结构的影响。结果 CGMP与Ara h1、Ara h2间存在次级键(盐桥、氢键、范德华力),部分作用于过敏原表位;MCGP致敏组血清中的花生蛋白特异性免疫球蛋白E(Specific immunoglobulin E, sIgE)、sIgG₁、sIgG_2a含量显著下降,白介素-4(interleukin-4, IL-4)、IL-5、转化生长因子-β(transforming growth factor-β, TGF-β)、组胺水平显著下降,肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)水平显著升高, MCGP中Ara h2的α-螺旋与β-折叠的比例改变。结论 CGMP能够改变Ara h2的结构,遮蔽花生过敏原表位,抑制sIgE、sIgG结合Ara h1、Ara h2,降低部分花生过敏原的免疫反应性。     

酪蛋白抗氧化肽的制备及初步分离

采用凝乳酶对牛乳酪蛋白酶解,以水解液对DPPH的清除率为指标,进行了酶解工艺优化。酶解液经超滤,Sephadex G-15凝胶层析柱和DEAE-52离子交换层析分离,得到了初步纯化。     

Dynamics of gelation,textural and microstructural properties of gelatin gels in the presence of casein glycomacropeptide

The aim of this work was to study the interaction between gelatin and casein glycomacropeptide (CMP) in the dynamic of gelation and the textural and microstructural properties of the mixed gels. Size particle, dynamic of gelation and textural and microstructural properties of CMP, gelatin and CMP-gelatin systems at pH 3.5 and pH 6.5 were determined. Size particle of gelatin increased by decreasing temperature from 35 °C to 5 °C, while no differences were observed in the size particle of CMP. At pH 6.5 the critical gelling concentration of gelatin was 1.5% and CMP did not gel, but the behavior of mixed systems was similar to gelatin. The more relevant result was observed at pH 3.5 since at concentrations in which CMP and gelatin did not gel on its own, the mixed systems gelled suggesting a synergistic effect.     

Heterogeneity of the kappa-casein glycomacropeptide and physiological activity of its fractions

A number of fractions with differing intensity of the inhibitory action on the gastric secretion were obtained by gel filtration in a column with biogels P-30 and P-2 of cappa-casein glycomacropeptide. A low-molecular fraction displays the greatest capacity for inhibiting gastric secretion. Desialism, heating and partial hydrolysis of glycomacropeptide with pepsin, trypsin and chemotrypsin used separately have no effect on its inhibitory activity. The resulting data suggest that the inhibitory action of glycomacropeptide is caused by a certain part rather than by entire molecule.     

Functional and biological activities of casein glycomacropeptide as influenced by lipophilization with medium and long chain fatty acid

This study determined the functional and different biological activities of casein glycomacropeptide (GMP) after conjugation with fatty acids. Medium (i.e. caproic, lauric and myristic acid) and long (i.e palmitic and stearic acid) fatty acids were conjugated to GMP at the available amino group. Functionalities of lipophilized GMP conjugates included foaming and emulsifying activities, and biological activities for bacterial growth inhibition, cell damage and anti-invasion. Greater lipophilization of GMP was achieved with medium chain fatty acids (p < 0.05), which resulted in reduced GMP solubility regardless of fatty acid conjugate. Foaming activity of GMP was lost after lipophilization, but emulsification activity of GMP was enhanced (p < 0.05). A parallel increase in growth inhibition of Salmonella spp. coupled with anti-invasion of Salmonella enteritidis (Inv A) into mammalian cells was induced by lipophilization of GMP with long chain fatty acid. Our results show that GMP modified by lipophilization with specific fatty acids provides improved functionality as a surfactant with enhanced antibacterial activity towards gram negative bacteria.