Denaturation of Bovine Serum Albumin (BSA) and Ovalbumin by High Pressure, Heat and Chemicals

Both native and denatured protein samples were examined by determining fluorescence and specific rotation, and by polyacrylamide gel electrophoresis (PAGE) and differential scanning calorimetry (DSC). Denaturation of ovalbumin by pressure was much less than by heat or by the chemical denaturants. Ovalbumin was denatured under high pressure, as confirmed by the decrease in its a-helical content to 72% and DSC endothermic enthalpy to 61%, but it showed no change in the PAGE pattern. With bovine serum albumin decrease in fluorescence was observed after denaturation by chemicals, but it did not change under high pressure.     

High pressure unfolding of ovalbumin

Summary The effects of high pressure processing of ovalbumin have been investigated. After treatment with pressures in excess of 400 MPa at pH 6.5, circular dichroism (CD) and Fourier transform infra-red spectroscopy (FTIR) spectroscopy showed limited irreversible changes in secondary structure. Fluorescence and derivative spectroscopy as well as fluorescence-quenching experiments indicated greater solvent exposure of aromatic residues in pressure-treated protein. Pressure treatment also caused enhanced binding of anilino-1-naphthalene-8-sulphonic acid (ANS). The pressure necessary to cause these changes was lower at low pH. These data indicate a pressure-induced molten-globule formation. The pressurized protein may be structurally similar to forms of the protein found at acid pH or as intermediates in protein folding.     

鸡蛋清中三种主要蛋白质的连续化提取分离研究

对鸡蛋清进行微滤、超滤浓缩后,加入60%的饱和硫酸铵盐析,得到了卵白蛋白粗品;上清液用Q-sepharoseF.F.离子交换层析,分离得到溶菌酶和卵铁传递蛋白,从而实现了蛋清中三种蛋白质的连续化提取。     

鸡蛋清磷酸化蛋白质组鉴定与分析

为了系统地阐明鸡蛋清磷酸化蛋白质的结构和功能特性,本研究采用蛋白质组学策略对鸡蛋清的磷酸化修饰蛋白质组进行鉴定与分析。鸡蛋清酶解产物首先经固定化金属螯合亲和层析分离富集磷酸肽,再经纳升液相色谱-串联质谱分析和鉴定。通过数据库检索匹配,共鉴定出33 个特异性磷酸化修饰多肽,包含41 个磷酸化修饰位点,归属于25 种鸡蛋清磷酸化蛋白。基序分析显示,大多数磷酸化修饰位点的序列特征为“S-X-E”;基因本体功能注释分析表明,鸡蛋清磷酸化蛋白质主要参与“生物调节”、“刺激反应”、“发育过程”等生物学过程,主要涉及“结合”、“催化”等分子功能。本研究结果将为鸡蛋清蛋白质相关研究提供关键的磷酸化修饰结构信息。     

鸡卵类黏蛋白过敏原性的在体与离体实验

通过观察卵类黏蛋白引起的豚鼠过敏性休克以及对致敏豚鼠离体肠平滑肌过敏性收缩反应（Schultz- Dale反应）的影响,研究鸡卵类黏蛋白的过敏原性,并初步建立其过敏实验的动物模型。结果显示：1）在实验 观察剂量下,1 mg/mL卵清白蛋白组豚鼠100%出现过敏性休克并死亡,全身性过敏反应呈极强阳性（4 分）; 卵类黏蛋白组豚鼠的反应与卵白蛋白组相似（P＞0.05）,大部分动物出现死亡,全身性过敏反应100%呈阳性 （2.04～3.68 分）;不同剂量的卵类黏蛋白引起的豚鼠全身过敏反应呈现出剂量依赖性,当致敏剂量为4 mg/mL 时,全身性过敏反应评分为3.68。2）卵类黏蛋白对Schultz-Dale反应的影响也与卵清白蛋白相似,使致敏的肠平滑 肌收缩活动明显加强;以2 mg/mL致敏时,豚鼠离体肠收缩力改变率最高,为538.5%。3）不同致敏剂量卵类黏蛋 白组豚鼠血清免疫球蛋白（immunoglobulin,Ig）G水平较对照组明显升高,且呈剂量依赖性。由此建立的卵类黏 蛋白在体与离体过敏实验的动物模型可用以检测卵类黏蛋白过敏原及评价其过敏原的消减效果。     

复合卵白蛋白-大豆分离蛋白对肉糜凝胶特性和微观结构的影响

运用卵白蛋白改善大豆蛋白与肉糜凝胶强度。通过在大豆分离蛋白中添加不同比例的卵白蛋白,应用于肉糜制品中,观察肉糜凝胶理化特性和微观结构,综合分析卵白蛋白和大豆分离蛋白复合凝胶在肉糜制品中的应用效果。结果表明：复合组显著改善产品的硬度、弹性、蒸煮损失、保水性（P＜0.05）,复合蛋白质量比为1∶1时,产品的脂肪聚集体达到最小,硬度得到改善,产品的弹性稳定且达到最大值,产品的可接受度最佳。     

温度对碱作用下卵白蛋白-葡萄糖体系色泽、理化特性及抗氧化活性的影响

探讨温度（4、25、37 ℃）对碱作用下卵白蛋白-葡萄糖体系色泽、理化特性及其抗氧化活性的影响。结果表明,随着孵育时间延长和温度升高,卵白蛋白-葡萄糖体系色泽不断加深,棕色产物以及荧光化合物含量不断增加,其美拉德反应中间产物含量在4 ℃和25 ℃时不断上升,而在37 ℃时先上升后下降,其pH值在整个孵育期间呈现出先上升后下降的趋势。傅里叶变换红外光谱分析表明,卵白蛋白和葡萄糖发生了缀合反应。氨基酸分析表明在碱诱导下卵白蛋白中的丝氨酸、组氨酸和半胱氨酸具有较高的反应性,其中反应性最高的是半胱氨酸。孵育后卵白蛋白-葡萄糖体系的抗氧化活性与卵白蛋白葡萄糖混合物相比有了显著提升,但随孵育时间的延长有轻微下降。     

Effect of ascorbic acid on the foaming and gelling of globular proteins

Foam expansion and foam stability of plasma and egg albumen proteins were enhanced in the presence of increasing concentrations of ascorbic acid (0.1–1.0%). BSA showed the greatest improvement in foaming properties following treatment with ascorbic acid, whilst foaming properties of egg albumen were improved to a limited extent. A combination of partial acid hydrolysis and treatment with 1% ascorbic acid was required to dramatically improve the foaming properties of bovine blood plasma. In the presence of sucrose, foam expansion of both native and ascorbic acid-treated blood plasma was decreased. In contrast the foam expansion of ascorbic acid-treated egg albumen was greater although this effect decreased slightly on incubation. the gelation of ascorbic acid treated proteins increased with increasing temperatures over 80–90°C, particularly for BSA and blood plasma, and with increasing concentrations of ascorbic acid. Ascorbic acid-treated proteins exhibited enhanced surface and exposed hydrophobicity and reduced numbers of sulphydryl groups indicating the involvement of these factors in foam and gel formation.