Effects of graded removal of lysine from an intravenously infused amino acid mixture on lactation performance and mammary amino acid metabolism in lactating goats

To investigate responses of milk protein synthesis and mammary AA metabolism to a graded decrease of postruminal Lys supply, 4 lactating goats fitted with jugular vein, mammary vein, and carotid artery catheters and transonic blood flow detectors on the external pudic artery were used in a 4 × 4 Latin square experiment. Goats were fasted for 24 h and then received a 9-h intravenous infusion of an AA mixture plus glucose. Milk yield was recorded and samples were taken in h 2 to 8 of the infusion period; a mammary biopsy was performed in the last hour. Treatments were graded decrease of lysine content in the infusate to 100 (complete), 60, 30, or 0% as in casein. Lysine-removed infusions linearly decreased milk yield, tended to decrease lactose yield, and tended to increase milk fat to protein ratio. Milk protein content and yield were linearly decreased by graded Lys deficiency. Mammary Lys uptake was concomitantly decreased, but linear regression analysis found no significant relationship between mammary Lys uptake and milk protein yield. Treatments had no effects on phosphorylation levels of the downstream proteins measured in the mammalian target or rapamycin pathway except for a tended quadratic effect on that of eukaryotic initiation factor 2, which was increased and then decreased by graded Lys deficiency. Removal of Lys from the infusate linearly increased circulating glucagon and glucose. Removal of Lys from the infusate linearly decreased arterial and venous concentrations of Lys. Treatments also had a significant quadratic effect on venous Lys, suggesting mechanisms to stabilize circulating Lys at a certain range. The 2 infusions partially removing Lys resulted in a similar 20% decrease, whereas the 0% Lys infusion resulted in an abrupt 70% decrease in mammary Lys uptake compared with that of the full-AA mixture infusion. Consistent with the abrupt decrease, mammary Lys uptake-to-output ratio decreased from 2.2 to 0.92, suggesting catabolism of Lys in the mammary gland could be completely prevented when the animal faced severe Lys deficiency. Mammary blood flow was linearly increased, consistent with the linearly increased circulating nitric oxide by graded Lys deficiency, indicating mechanisms to ensure the priority of the mammary gland in acquiring AA for milk protein synthesis. Infusions with Lys removed increased mammary clearance rate of Lys numerically by 2 to 3 fold. In conclusion, the decreased milk protein yield by graded Lys deficiency was mainly a result of the varied physiological status, as indicated by the elevated circulating glucagon and glucose, rather than a result of the decreased mammary Lys uptake or depressed signals in the mTOR pathway. Mechanisms of Lys deficiency to promote glucagon secretion and mammary blood flow and glucagon to depress milk protein synthesis need to be clarified by future studies.     

南美白对虾肝胰腺蛋白提取方法优化及无水保活环境胁迫对细胞凋亡蛋白的影响

为提高Western Blotting结果的可靠性,以蛋白溶出率和凝胶电泳蛋白条带完整性为指标,比较提取液种类、研磨方式、酶抑制剂种类及其体积分数和提取时间对南美白对虾肝胰腺蛋白提取效果的影响。采用优化后的提取方法获得高质量蛋白样品,并采用Western Blotting法分析无水环境胁迫后南美白对虾肝胰腺组织中细胞凋亡信号通路相关蛋白的表达水平。结果表明：RIPA裂解液作为提取溶剂所得的蛋白溶出率高于水提和磷酸盐缓冲液,电动匀浆和液氮研磨所得蛋白条带更完整,4%蛋白酶磷酸酶混合抑制剂能有效抑制肝胰腺内源酶引起的蛋白降解;采用Western Blotting法分析无水保活期间南美白对虾肝胰腺蛋白,发现低温诱导休眠的同时会引起细胞轻微凋亡,且凋亡水平呈应激时间依赖性增加,环境胁迫解除后有所回调。     

Preparation and Applications of Soybean Residue CNF Films

In this study, soybean residues were treated with HCl and soybean residue cellulose was extracted, which was used to prepare cellulose nanofiber (CNF) using the high-pressure homogenization method. The maximum yield of CNF, the reaction temperature, reaction time, and HCl concentration were optimized. The optimum HCl concentration for acid treatment was 6%, the reaction time was 60 min, the reaction temperature was 80℃, and the maximum yield of soybean residue cellulose was 78.8%. The different CNF films were then prepared; the color, mechanical property, and light transmittance of the CNF films were studied. Compared to the properties of the CNF film prepared with the soybean residue cellulose by high-pressure homogenization 15 times (HGT-15 film), the mechanical properties of the CNF film with soybean residue cellulose by decolorizing treatment decreased, but the light transmittance increased. The film prepared by adding HGT-15 CNF to whey protein was investigated for its mechanical property, light transmittance, and solubility. Unlike the pure whey protein film, addition of 2.0% CNF to the whey protein enhanced the mechanical property and water vapor transmission rate (WVT) of the film. With the increase in CNF content, the solubility of the whey protein film decreased, and then stabilized.     

再生蛋白纤维研究进展

再生蛋白纤维是一种性能优良,应用广泛的新型绿色环保材料,可分为再生植物蛋白质纤维与再生动物蛋白质纤维两大类。文章介绍了再生蛋白纤维的发展历程,以及以大豆蛋白、牛奶蛋白、玉米蛋白等为原料加工再生蛋白纤维的制备、性能及其应用,并对其今后的发展前景进行了展望。     

谷氨酰胺转移酶对大豆分离蛋白膜性能的影响

研究注模前酶作用时间对谷氨酰胺转移酶(TGase)改性大豆分离蛋白(SPI)膜性能的影响。在注模之前,将TGase(8U/g SPI)加入到成膜溶液中,分别在磁力搅拌下作用0、30、60、120min,然后注模成膜。利用质构仪检测蛋白膜的机械性能,结合哈克流变仪的动态黏弹实验及SDS-PAGE 实验进一步分析。注模前适度的酶作用(≤60min)在一定程度上有利于TGase 改性的SPI 膜机械强度的提高,特别是抗拉强度(TS)值;但是,时间不宜过长,因为注模前的酶作用也会诱导SPI 蛋白组分的聚沉反应,从而降低成膜溶液中可溶解蛋白的含量。结果表明,TGase改性SPI 膜时,一方面会诱导蛋白交联;另一方面,交联过多又会导致沉淀;在利用TGase 提高SPI 膜的机械性能时如何把握两者之间的关系,在交联的同时抑制酶促聚沉非常重要。     

酶法从全酯大豆中同时制备大豆油和大豆水解蛋白工艺的研究

研究了酶法从全脂大豆中同时制各大豆油和大豆水解蛋白的工艺，水提过程的最佳工艺条件为固液比１：１０，温度４４℃，ｐＨ７．７０和提取时间３６ｍｉｎ。含油大豆蛋白进行两次有控制的酶解，得到等电点可溶大豆水解蛋白（ＩＳＳＰＨ）和稳定性低的乳化油。确定了转相法破乳的工艺条件，从乳化油分离得到纯度较高的大豆油。水解蛋白和油的得率分别达到７４％和６６％。探讨了等电点或可溶大豆水解蛋白的功能性质及其在食品中的应用。     

椰子分级蛋白制备的研究

通过改良的Osboren法对脱脂椰子粉进行分级提取,得到清蛋白、球蛋白、醇溶蛋白和谷蛋白,并对4种椰子蛋白进行氨基酸组成分析和SDS-PAGE电泳分析。在单因素实验基础上,利用正交实验对4种椰子蛋白提取条件进行优化。结果表明,4种椰子蛋白最佳提取工艺条件为:在液料比10∶1、提取时间4 h、提取温度50℃条件下,清蛋白提取率为49. 05%,纯度为57. 1%;在液料比10∶1、提取时间4 h、提取温度50℃条件下,球蛋白提取率为46. 65%,纯度为70. 8%;在液料比10∶1、提取时间5 h、提取温度55℃、乙醇体积分数75%条件下,醇溶蛋白提取率为18. 01%,纯度为32. 7%;在液料比10∶1、提取时间4 h、提取温度50℃、碱溶p H 10. 0条件下,谷蛋白提取率为26. 78%,纯度为70. 4%。椰子清蛋白、球蛋白和谷蛋白中谷氨酸、精氨酸和天冬氨酸的含量明显高于其他氨基酸。除了清蛋白有2条谱带相对分子质量较大,其他3种椰子蛋白相对分子质量都相对较小,整体来说椰子蛋白是一种较为优质的蛋白质。     

蛋清蛋白对豌豆淀粉凝胶化及凝胶特性的影响

为探究蛋清蛋白对豌豆淀粉凝胶化及凝胶特性的影响。分别以0%、3%、6%、9%和12%的蛋清蛋白替代豌豆淀粉,研究蛋清蛋白对豌豆淀粉糊化特性、热特性、静态流变、动态流变、凝胶质构及水分子状态的影响。结果表明：蛋清蛋白以浓度依赖的方式影响豌豆淀粉的糊化特性,峰值黏度、谷值黏度、崩解值、最终黏度、回生值均随蛋清蛋白添加量增加而降低,而糊化温度则升高。蛋清蛋白对豌豆淀粉凝胶化温度影响不显著,但降低了淀粉的凝胶化焓值。添加不同量蛋清蛋白的豌豆淀粉糊均表现为假塑性流体特征,Herchel-Bulkley模型能够很好地拟合其静态流变行为,稠度指数和屈服应力均随蛋清蛋白添加量增加而降低。添加蛋清蛋白降低了豌豆淀粉糊体系的黏弹性及凝胶的硬度、强度和可塑性。蛋清蛋白对豌豆淀粉凝胶中自由水含量影响不显著,但使结合水含量增加而不可流动束缚水含量降低。     

Development of a homogeneous competitive immunoassay for phosphorylated protein antigen based on the enhanced fluorescence resonance energy transfer technology

We describe a homogeneous competitive immunoassay for a phosphorylated protein antigen. The assay takes advantage of the enhanced fluorescence resonance energy transfer (FRET) technology, which has a unique characteristic that the FRET signal is increased by the specific interaction of two fluorolabeled leucine zippers. We chose extracellular signal-regulated kinase (ERK) as a model antigen and constructed two molecular probes in which either anti-phosphorylation site antibody or the antigen peptide was chemically conjugated to the enhanced FRET probes. While these molecular probes indicated sufficient FRET signal without antigen, they displayed a significant change in the fluorescent spectrum by mixing with phosphorylated antigens. With this competitive enhanced FRET immunoassay, a phosphorylated ERK concentration within the range from 15 nM to 250 nM could be determined. Because the assay is very simple, it would be applied to not only in vitro assay but also in vivo detection of protein phosphorylation.     

热处理和碱处理对可食性大豆分离蛋白膜性能的影响

研究了热处理和碱处理对可食性大豆分离蛋白(SPI)膜性能的影响。对成膜液进行适当的加热和调节pH可以提高SPI膜的抗拉强度(TS)和伸长率(E),降低水蒸气透过系数(WVP)。调节成膜液的pH到9,在70℃加热20min,所得到的膜机械性能和阻湿性能最好。