α-乳白蛋白(bovine α-lactalbumin,BLA)可引发过敏反应,且与肠道菌群有关,通过不同蛋白质修饰技术对BLA进行改性,探究其对致敏性和人肠道菌群的影响。以糖基化、磷酸化及乙酰化修饰的BLA为研究对象,采用电泳、光谱学和ELISA等技术研究其结构和IgE结合能力的变化,并运用16S rRNA测序方法分析人肠道菌群的结构及组成。结果表明:BLA经磷酸化、糖基化和乙酰化修饰后,其结构发生了改变;修饰后BLA的IgE结合能力逐渐降低;16S rRNA高通量测序结果表明,与BLA相比,修饰后的BLA在门水平上降低拟杆菌门的相对丰度,增加厚壁菌门的相对丰度;在属水平上,其能明显增加巨单胞菌属和罗姆布茨菌属的相对丰度,而降低普雷沃氏菌属和粪杆菌属的相对丰度。研究结果表明,糖基化、磷酸化和乙酰化修饰通过破坏BLA的结构降低其致敏性,且影响人肠道微生物的结构,影响程度为乙酰化>糖基化>磷酸化。
Bovine α-lactalbumin (BLA) can induce allergic reactions and is related to intestinal microbiota.The effects of BLA modification on allergenicity and human intestinal microbiota were investigated by modifying BLA with different protein modification techniques.In this study,glycationed,phosphorylated and acetylated BLA was used as the object of study.Electrophoresis,spectroscopy and ELISA were used to study the changes of its structure and IgE binding ability.16S rRNA sequencing was used to analyze the structure and composition of human intestinal microbiota.The results showed that the structure of BLA was changed by phosphorylation,glycation and acetylation.After modification,the IgE binding ability of BLA decreased gradually.High-throughput sequencing of 16S rRNA showed that,compared with BLA,the modified BLA decreased the relative abundance of Bacteroidota and increased the relative abundance of Firmicutes at the phylum level.At the genus level,it significantly increased the relative abundance of Megamonas and Romboutsia,but decreased the relative abundance of Prevotella and Faecalibacterium.The results showed that glycation,phosphorylation and acetylation reduced the allergenicity of BLA by destroying its structure and affected the structure of human intestinal microbiota to the extent of acetylated>glycation>phosphorylation
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