摘要
目的通过测定转人α-乳清白蛋白基因的克隆牛所表达的重组人α-乳清白蛋白在模拟胃肠液及小型猪胃肠液中的消化时间,并与人α-乳清白蛋白标准品在上述消化液中的消化时间相比较,判定该重组蛋白的消化稳定性,为其致敏性评价提供基础资料。方法根据《转基因生物及其产品食用安全检测——模拟胃肠液外源蛋白质消化稳定性试验方法》消化人α-乳清白蛋白标准品和重组人α-乳清白蛋白,以十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白免疫印迹法(Western blotting)分析消化后的样品。结果重组人α-乳清白蛋白在模拟胃液中0 s即被完全消化,模拟肠液中10 min内完全消化,在小型猪胃液和肠液中均在20 min内被完全消化。虽然完全消化的时间存在差异,但均在20 min内。重组蛋白与人α-乳清白蛋白标准品的消化稳定性相似。结论重组人α-乳清白蛋白在模拟胃肠液及小型猪胃肠液中均不具有消化稳定性。
Objective To judge the stability and digestibility of human α-lactalbumin derived from recombinant bovine.To compare the digestion time for the recombinant human α-lactalbumin with that for standard human α-lactalbumin in simulated gastric and intestinal fluid or in gastric and intestinal fluid extracted from miniature swine.This would provide basic materials for assessing the allergenicity of recombinant proteins.MethodsThe recombinant protein and human α-lactalbumin standard substance were digested according to the national standard method Food safety detection of genetically modified organisms and derived products——Method of target protein digestive stability in simulative gastric and intestinal fluid.The samples after digestion were analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis（SDS-PAGE） and Western blotting.ResultsThe recombinant protein samples were digested completely within 0 s in simulated gastric fluid;10 min in simulated intestinal fluid;and 20 min in gastric and intestinal fluid of miniature swine.The digestion time was all less than 20 min for recombinant protein samples,which was similar for human α-lactalbumin standard substance,although some differences were existed.ConclusionThere was no digestion resistance of the recombinant protein in simulated gastric and intestinal fluid or in gastric and intestinal fluid extracted from miniature swine.
出处
《中国食品卫生杂志》
北大核心
2011年第3期205-211,共7页
Chinese Journal of Food Hygiene
基金
国家高科技研究发展计划(863计划)(2007AA100506)
