壁虎仿生酶解产物的凝胶分析系统比较研究

Comparative Study on Gel Analysis System of Biomimetic Enzymatic Hydrolysis Products of Gekko

关键词:  壁虎  仿生酶解  小分子肽  Tricine-SDS-PAGE法

Key words:  gekko  biomimetic enzymatic hydrolysis  small molecular peptide  Tricine-SDS-PAGE method

摘要:

目的 优选适合分离壁虎小分子肽的凝胶分离体系，为提高壁虎多肽的生物利用度和小分子肽分离分析提供实验依据。方法 采用仿生酶解法体外酶解壁虎蛋白质，采用福林酚比色法测定仿生酶解前后肽含量的变化，分别采用SDS-PAGE凝胶系统和Tricine-SDS-PAGE凝胶系统比较分析仿生酶解前后肽数量及分子量的变化。结果 壁虎经仿生酶解处理后，小分子肽含量增加，分子量降低，且Tricine-SDS-PAGE凝胶系统所能分离的条带数量明显多于SDS-PAGE凝胶系统。结论 Tricine-SDS-PAGE电泳三层凝胶系统更适用于分离壁虎小分子肽。

Abstract:

OBJECTIVE To optimize the gel separation system suitable for the separation of gekko small molecular peptides and to provide experimental basis for improving the bioavailability of gekko peptides and the separation and analysis of small molecular peptides. METHODS Biomimetic enzymatic hydrolysis was used to hydrolyze gekko protein in vitro, and folin-ciocalteu colorimetry was used to determine the changes in peptide content before and after biomimetic enzymatic hydrolysis. SDS-PAGE gel system and Tricine-SDS-PAGE gel system were respectively used to compare and analyze the changes in the number and molecular weight of peptide before and after biomimetic enzymatic hydrolysis. RESULTS After the gekko was treated by biomimetic enzymatic hydrolysis, the content of small peptides increased while the molecular weight decreased. Moreover, the number of bands separated by Tricine-SDS-PAGE gel system was significantly more than that of SDS-PAGE gel system. CONCLUSION The three-layer gel system of Tricine-SDS-PAGE electrophoresis is more suitable for the separation of gekko small molecular peptides.