食品科学2016，Vol.37Issue(19)：64-69,6.DOI:10.7506/spkx1002-6630-201619011
基于质谱检测转基因生物外源蛋白质的消化稳定性
Mass Spectrometry-Based Analysis of Digestive Stability of Target Protein in Genetically Modified Organism
摘要
Abstract
Digestive stability analysis of exogenous protein is one of the important indexes of geneticaly modified organisms (GMO) safety assessment. In the present study, we established a novel assessment assay for protein digestive stability based on label-free quantification of mass spectrometry using bovineβ-lacto globulin (BLG), bovine serum albumin (BSA) and soybean trypsin inhibitor (STI) as standard proteins. Protein samples were treated with simulated gastric/intestinal fluids (SGF/SIF), and then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The target bands were then cut out and trypsinized. The extracted peptides were analyzed by nano liquid chromatography-electronic spray ion-mass spectrum/mass spectrum and identified by Mascot software. By calculating the ratio between the matched peptide numbers of the target protein before and after SGF/SIF treatment, the digestibility of the target protein was estimated. When the ratio was lower than 0.50, the target protein was considered digestible. This newly developed assay was applied to Cry1Ab/1Ac, the exogenous protein of the geneticaly modified rice‘Huahui No. 1’. The results showed that the ratio was lower than 0.50 after digestion in SGF for 2 minutes, while it decreased to less than 0.50 after digestion in SIF for 15 seconds, indicating that it is very liable to be digested. Our study provides a novel MS-based method for digestive stability analysis of target proteins from genetically modified organisms, which is easy to operate with no need for specific antibody.关键词
转基因生物/消化稳定性/质谱/外源蛋白Cry1Ab/1Ac/无标记定量Key words
genetically modified organisms/digestive stability/mass spectrometry/exogenous protein Cry1Ab/1Ac/lable-free quantification分类
轻工纺织引用本文复制引用
毛劼,孙兴,程娟献,王心正,赵永强,王红霞,何昆,夏晴..基于质谱检测转基因生物外源蛋白质的消化稳定性[J].食品科学,2016,37(19):64-69,6.基金项目
国家科技重大专项（2014ZX08011007；2016ZX08011007）；国家重大科学仪器设备开发专项 （）
