大豆粘性功能肽的构效关系、粘性机理及对骨折愈合作用的研究

项目名称： 大豆粘性功能肽的构效关系、粘性机理及对骨折愈合作用的研究

项目编号： No.21476103

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 有机化学

项目作者： 沈新春

作者单位： 南京财经大学

项目金额： 90万元

中文摘要： 从大豆蛋白中制备安全、高效、无毒的医用粘性肽具有重要的社会和经济意义，而阐明其构效关系和粘接作用机制是目前亟待解决的问题。本研究将采用独创的限制酶解技术，以在体和离体动物骨骼为实验对象进行粘接性能的评价，从大豆亚基蛋白中筛选出高粘接活性的功能肽；用质谱技术、圆二色光谱（CD）、红外光谱（2D-IR）、差示扫描量热法（DSC）等方法对高粘接活性肽进行结构鉴定与表征，运用氨基酸变异技术对粘性肽进行构效关系和表征的研究，确定其粘性作用的功能区域（关键位点），在分子水平阐明其粘性机理；通过粘性肽与动物骨骼的吸附、浸润和粘附性能的研究，阐明其粘接作用机理；以成骨细胞（MC3T3-E1）和动物骨折模型为研究对象，采用多种研究方法和角度从不同水平阐明粘性肽在骨折愈合中的作用。本研究将阐明大豆粘性肽的构效关系、粘接机理及其在骨折愈合中的作用，为大豆蛋白高值化加工利用和新型医用生物粘合剂的开发奠定理论基础。

中文关键词： 大豆蛋白；医用粘性肽；限制酶解；构效关系；骨折愈合

英文摘要： Safe, efficient, non-toxic medical bio-adhesive peptide from soy protein has important social and economic significance. However, how to prepare such peptides with high-added value from soy protein, and its structure-activity relationships (SAR) are unknown, and its adhesive mechanisms are not yet fully elucidated. In the present study, highly adhesive peptides from the subunits of soybean protein will be obtained by using a novel limited proteolysis and separation. The shear and tensile strengths of adhesive peptides will be investigated using both in vitro and in vivo evaluation methods, and their adhesive structures will be characterized using functional proteomics, circular dichroism (CD), infrared spectroscopy (2D-IR), differential scanning calorimetry (DSC) and so on. In addition, we will further confirm the SAR and characterization of adhesive peptides, identify the functional regions for affecting its adhesive strength, and clarify the adhesive molecular mechanism using replacement (mutation) of amino acid method and by determining of the correlations between adsorption, infiltration, and adhesion properties of highly adhesive peptides with the substrate (animal bones). Moreover, we will incubate the osteoblasts (MC3T3-E1) cells with adhesive peptides and determine the effect of its peptides on the proliferation, differentiation, and safety of cells. We will establish the rabbit models of mandibular fractures and glue by the adhesive peptides, and then observe ossification and perform biological evaluation during the process of fracture healing. Taken together, this study will have significant implications in understanding important SAR and molecular mechanisms of adhesive peptides in the process of fracture healing, and provide insightful theoretical basis for developing the high-value processing and utilization of soybean protein as new medical bio-adhesives.

英文关键词： Soybean protein;Medical adhesive peptides;Limited proteolysis;Structure-activity relationship （SAR）;Fracture healing