真菌Penicillium sp. MO3 产生的抗菌肽AF2的分离纯化和性质的研究

从真菌Penicillium sp. M03发酵液中提取到一种具有抗菌活性的肽类物质AF2,并对其性质进行初步研究. 真菌发酵液经纱布过滤,离心后经10000D超滤膜超滤,冻干,以纯水溶解经Sephadex LH-20,DEAE Sepharose及Sephadex G-15进行分离纯化得到纯净物质,并对其基本性质进行研究. 结果证明：AF2具有典型的肽类物质特征反应,分子量约为1155,含有12种常见氨基酸,对酸碱,胰蛋白酶,β-内酰胺酶稳定,对热有较好的耐受性.     

瑞士乳杆菌发酵制备牦牛乳酪蛋白源性降血压肽的工艺研究

目的:以牦牛乳源酪蛋白为原料,利用瑞士乳杆菌,对发酵法制备降血压肽的工艺进行初步探讨.方法:试验首先采用凝乳酶沉淀法从耗牛乳中提取酪蛋白,依据瑞士乳杆菌的发酵特点,按5％(体积比)接菌量进行发酵制备降血压肽.然后并以降血压肽对血管紧张素转换酶(ACE)的抑制率为指标,主要探讨了底物浓度、pH值、发酵时间三因素对瑞士乳杆菌发酵法制备降血压肽工艺的影响.结果:在接菌量为5％,其他三因素各设置五个水平进行工艺优化,最优工艺条件为:底物浓度4％、发酵pH值8.0、发酵时间16 h.此工艺条件下ACE抑制率可达约68％.结论:通过实验说明底物浓度、pH值、发酵时间等参数的控制对发酵酪蛋白制备抑制肤很重要,此试验数据为后续进一步完善制备降血 压肤的实验奠定了重要的基础.     

抗菌多肽的研究进展

抗菌多肽是在生物体中具有抗菌活性的肽类物质的总称,是无数动植物天然免疫系统的重要组成部分,具有抗菌谱广、作用强且迅速、不易产生耐药等优点。抗菌多肽已经成为药物开发的研究热点。本文简要介绍抗菌多肽的分类、作用和作用机理,并对抗菌多肽的应用前景及其存在问题进行分析。     

Enhancement of transfection efficiency for HeLa cells via incorporating arginine moiety into chitosan

Arginine-rich peptides have attracted considerable attention due to their distinct internalization mechanism. It was reported that arginine and guanidino moieties were able to translocate through cell membranes and played a critical role in the process of membrane permeation. In this work, arginine was conjugated to the backbone of chitosan to form a novel chitosan derivative, arginine modified chitosan (Arg-CS). Arg-CS/DNA complexes were prepared according to the method of coacervation process. The physicochemical properties of Arg-CS and Arg-CS/DNA complexes were characterized and the transfection activity and efficiency mediated by Arg-CS/DNA complexes were investigated taking HeLa cells as target cells. Arg-CS was characterized by FTIR and 13C NMR. Arg-CS/DNA polye- lectrolyte complexes were investigated by agarose gel retardation, dynamic light scattering (DLS) and atomic force microscopy (AFM). The results revealed that the Arg-CS/DNA complexes started to form at N/P ratio of 2:1, and the size of particles varied from 100 to 180 nm. The cytotoxicity of Arg-CS and their complexes with plasmid DNA were determined by MTT assay for HeLa cells, and the results suggested that Arg-CS/DNA complexes were slightly less toxic than Arg-CS. Moreover, the derivative alone and their complexes showed significantly lower toxicity than PEI and PEI/DNA complexes, respectively. Taking HeLa cells as target cells and using pGL3-control as reporter gene, the luciferase expression mediated by Arg-CS was greatly enhanced to about 100 folds compared with the luciferase expression mediated by chitosan at different pH media. These results suggest that Arg-CS is a promising candi- date as a safe and efficient vector for gene delivery and transfection.     

鲷鱼鳞多肽的酶法制备及抗氧化活性研究

以鲷鱼鳞为原料,经物理方式处理后,采用单因素实验考察液料比、pH值、温度、酶底比及酶解时间对鲷鱼鳞酶解液水解度和DPPH自由基清除率的影响.结合响应面优化设计对酶解条件进行优化,结果在液料比为28∶1、酶底比7.0%、pH值7.3、50℃、酶解6 h条件下,所得酶解液水解度为10.35%,DPPH自由基清除率为82.33%.DPPH自由基清除、羟基自由基清除、还原力、金属螯合测试表明,酶解产物的抗氧化活性与浓度呈现剂量依赖性,具有抗氧化活性.分子量测定及氨基酸组成分析表明,酶解产物含有大量的寡肽,相对分子质量分布区间为180~1 000 u,其富含甘氨酸、丙氨酸、羟脯氨酸,且含有多种人体必需的氨基酸.     

抗菌肽的研究进展

抗菌肽具有分子量小、热稳定性好、抗菌谱广等特点,介绍了抗菌肽的组成结构、分类、生物学特性、抗病机制、基因工程、发展前景和存在的问题。     

鳕鱼皮复合肽的制备工艺研究

采用木瓜蛋白酶和风味酶构成的复合酶对鳕鱼皮蛋白进行水解,通过单因素和正交试验确定复合酶水解的最佳条件为：木瓜蛋白酶和风味酶复合比为3：1,加酶量为40 000 U/g,水解温度45℃,料液比为1：2,水解pH为6.5,水解时间为4.5 h。最佳条件下鳕鱼皮蛋白水解度可达22.45%。     

燕麦多肽数据库及降血糖活性的虚拟筛选

为了深入研究燕麦多肽中可能发挥降血糖功能的活性多肽分子，本文首先从文献中调研了从燕麦中提取鉴定得到的多肽，构建了对应的燕麦多肽数据库，并基于DPP4蛋白对多肽数据库进行了虚拟筛选.随后，针对筛选获得的6个多肽分别进行了100 ns的分子动力学模拟.从模拟之后稳定结合的构象分析了不同多肽分子与DPP4的相互作用信息，并分别计算了不同多肽分子与DPP4的结合自由能.结果表明，从燕麦多肽数据库中筛选得到的多肽可以与DPP4蛋白稳定结合，其中2个多肽分子与DPP4的亲和力相对较强.本文得到的多肽分子可以作为后续DPP4抑制剂设计和改造的先导分子，燕麦多肽数据库也可用于研究燕麦的其他生物学功能.