利用温敏性凝胶的缓释作用促进变性溶菌酶复性

为了开发温敏凝胶在蛋白质复性方面的方法，利用N-异丙基丙烯酰胺温敏凝胶具有的缓释作用，对变性蛋白质进行了复性．考察不同合成条件下凝胶的性能，结果表明，在制胶温度为22℃、凝胶单体浓度为10％、交联度为5％制得的凝胶，具有较好的机械强度、较大的溶胀倍率、缩水倍率及良好的温敏性能．凝胶的缓释作用对变性还原溶茵酶辅助复性的影响表明，在复性酶终浓度为0．5mg／mL时，凝胶辅助蛋白质的复性收率可达68％，比直接稀释复性（收率为57％）高19％．特别是对高浓度蛋白质的复性（1．5mg／mL），凝胶辅助复性也获得了很好的效果，复性收率高出直接稀释复性37％，表现出温敏凝胶在实际蛋白质复性中的应用前景．     

溶菌酶脂质体的制备及其对生物膜的剥离作用

采用逆向蒸发法制备了稳定的溶菌酶脂质体.在不锈钢表面培养出稳定生物膜后,分别利用溶菌酶和溶菌酶脂质体对其进行剥离.运用Zeta电位仪、扫描电子显微镜(SEM)和红外光谱(FT-IR)对脂质体和生物膜进行表征.结果表明制备的脂质体平均粒径为80~100 nm,包封率为82.4%.相同浓度下溶菌酶及其脂质体对混合菌种形成的生物膜剥离效率分别达到62.4%和86.5%.溶菌酶脂质体在24h内对生物膜和水体中微生物去除率分别达到89.6%和99.6%.因此,溶菌酶脂质体能够有效控制不锈钢表面生物膜污染风险.     

定量负荷对唾液溶菌酶含量影响初探

为探讨定量负荷对大学生唾液溶菌酶(lysozyme缩写LZM)含量的影响,我们对168名大学生LZM含量进行了测定分析。实验结果表明,在静态时,体育系学生LZM含量低于普系学生;定量负荷后即刻体育系学生LZM含量明显增加;不同运动专项的体育学生LZM含量亦有差别,球类专项学生其含量低于田径专项。     

巴氏毕赤酵母SMD1168表达人溶菌酶的发酵条件

研究用毕赤酵母SMD1168菌株(MutS,HIS4+)表达人溶菌酶(HLZ)的发酵条件。此菌株具有对胞外分泌的外源蛋白不降解、对反应器氧传递效率的限制不敏感的特性,有利于在常规反应器条件下的过程放大。在培养基中添加复合维生素,在流加甲醇溶液中添加山梨醇或甘露醇可维持细胞正常的生长和代谢,并表达高活性的HLZ。采用恒溶解氧的方式流加甘油溶液以提高细胞密度,在一段时间内使细胞处于碳源半饥饿状态后,使用恒速流加甲醇溶液诱导HLZ表达。HLZ的体积生成速率达到9.6mg/(L·h)左右,上清液中HLZ的含量约为1.6g/L,发酵周期为110h。     

Expression of recombinant human lysozyme in the milk of transgenic mice

Human lysozyme is a 130-aa (amino acid) alkaline polypeptide, and has both anti-bacterial and anti-viral properties which make it an important component of human natural immunity system. As a first step toward the ultimate goal of improving the anti-bacterial properties of bovine and ovine milk, a transgenic mouse that contains the genomic DNA sequence of the human lysozme gene has been generated for the first time. From 83 mice generated by microinjection, a total of 6 positive transgenic mice were identified by PCR and Southern blot. F1 mice positive for transgene in lines were also detected by PCR. This shows that transgene could be transmitted from founder transgenic mice to their offspring. Recombinant human lysozyme (rHlys) was found in the whey of 3 female positive transgenic mice by Western blot. The highest concentration of rHlys for transgenic mice was 0.2mg/mL. The antibacterial activity of the whey for transgenic mice was highly enhanced up to 0.4 times as much as that of human, while that of non-transgenic mouse was very low. Although the lysozyme activity of transgenic mice is still lower than that of human, the rHlys exhibits the same specific activity as that of human lysozyme. It provides a strong basis for further studies into the possible application of rHlys express in mammary gland.     

Biological activity and pathological implications of misfolded proteins

The physiological metabolism of proteins guarantees that different cellular compartments contain the appropriate concentration of proteins to perform their biological functions and, after a variable period of wear and tear, mediates their natural catabolism. The equilibrium between protein synthesis and catabolism ensures an effective turnover, but hereditary or acquired abnormalities of protein structure can provoke a premature loss of biological function, an accelerated catabolism and diseases caused by the loss of an irreplaceable function. In certain proteins, abnormal structure and metabolism are associated with a strong tendency to self-aggregation into a polymeric fibrillar structure, and in these cases the disease is not principally caused by the loss of an irreplaceable function but by the action of this new biological entity. Amyloid fibrils are an apparently inert, insoluble, mainly extracellular protein polymer that kills the cell without tissue necrosis but by activation of the apoptotic mechanism. We analyzed the data reported so far on the structural and functional properties of four prototypic proteins with well-known biological functions (lysozyme, transthyretin, β2-microglobulin and apolipoprotein AI) that are able to create amyloid fibrils under certain conditions, with the perspective of evaluating whether the achievement of biological function favors or inhibits the process of fibril formation. Furthermore, studying the biological functions carried out by amyloid fibrils reveals new types of protein-protein interactions in the transmission of messages to cells and may provide new ideas for effective therapeutic strategies. Received 9 November 1998; received after revision 15 January 1999; accepted 15 January 1999     

偶氮类食品着色剂诱惑红与蛋溶菌酶的相互作用研究

采用荧光光谱、紫外吸收光谱和计算机模拟分子对接研究了偶氮类食品着色剂诱惑红与蛋溶菌酶的相互作用.讨论了诱惑红对蛋溶菌酶荧光的猝灭机理,属于静态猝灭.测定了诱惑红与蛋溶菌酶反应体系的结合常数和结合位点数,根据热力学参数焓变(ΔH>0)和熵变(ΔS>0)可推断诱惑红和蛋溶菌酶的相互作用力为典型的疏水作用力.在298、304、310 K温度下,两者间的结合常数分别为17897、19865、23267.     

杨树叶对鸡日粮中蛋白质和脂肪利用率的影响

研究了杨树叶对滨白蛋鸡蛋白质利用率和脂肪利用率的影响,结果表明:20%杨树叶可使滨白蛋鸡对日粮中蛋白质利用率提高7.80%,对日粮中脂肪利用率提高3.96% .用杨树叶干粉部分替代蛋鸡日粮可提高蛋鸡对蛋白质、脂肪等营养物质的利用率.     

盐酸溶液中溶菌酶对Q235钢的缓蚀作用

运用失重法、动电位极化曲线法、交流阻抗法研究了溶菌酶在盐酸溶液中对Q235钢的缓蚀作用和吸附行为,并用扫描电子显微镜(SEM)观察腐蚀表面的形貌.结果表明,溶菌酶是一种缓蚀效果显著的混合型缓蚀剂,缓蚀效率受温度影响较小,吸附行为属于自发的单分子层化学吸附,符合Langmuir等温吸附式.