双效型酵母细胞壁提取物及其对黄曲霉毒素B_1吸附特性研究

Double function yeast cell-wall extract and its capability of adsorbing aflatoxin B_1

本研究旨在利用麝香草酚裂解酵母细胞,同时以酵母膜脂为载体,与细胞壁共同组成具备抑制黄曲霉孢子萌发和吸附毒素能力的双效型细胞壁提取物。结果表明:通过流式细胞仪及荧光显微镜检测发现麝香草酚可以杀死酵母细胞;当使用500μg/m L麝香草酚制备其酵母细胞壁提取物,采用苯酚硫酸法分析发现制备物中β-葡聚糖的含量升高51.5%,而通过SDS-PAGE研究结果表明,胞内可溶性蛋白含量降低40%。这些结果表明麝香草酚可以诱导酵母细胞发生裂解,破坏其细胞膜的完整性,导致细胞壁上肽聚糖的结构发生变化、胞内内容物释放,增强了对黄曲霉毒素B_1的吸附能力。利用薄层层析(TLC)和液相色谱(HPLC)检测酵母细胞壁提取物对黄曲霉毒素的吸附情况,结果表明制备的提取物对黄曲霉毒素的吸附能力达到2.5μg/mg,对麝香草酚的载药量达到41.5μg/mg。本文可为粮食、饲料中控制黄曲霉毒素污染提供新的研究思路及理论依据。

In this study,we managed to produce a type of cell-wall extract with double functions i.e.spore inhibition and toxins adsorption by using thymol to anchor the phospholipid of yeast cells and lyse them. It was showed that thymol led to the yeast cell death with the observation by using the flow eytometry and fluorescence microscope.When yeast cell-wall extract by yeast cell was treated with 500 μg/mL thymol, the content of β-glucan was increased by 51.5 % in the cell-wall extracts by phenol -sulfuric ,and soluble protein in cells was decreased by 40% by SDS-PAGE.This indicated that thymol gave rise to tile lysis of yeast cell and destroyed the integrity of the cell membrane,the structural of peptidoglyean on the cell wall changed,yeast cell released their contentand and the adsorption capacity of AFB1 increased .The capability of cell-wall extract adsorbing AFB1 was 2.5 μg/mg by thin layer chromatography （ TLC ） and high performance liquid chromatography （ HPLC ）. And its thymol- loading capacity was 41.5 μg/mg. This study provided new ideas and theoretical basis for the control of aflatoxin contamination in food and feed.