【摘要】 在高产麦角硫因（ergothioneine,EGT）的食药用真菌筛选中，现有的EGT检测方法因其步骤繁琐、使用的试剂和设备昂贵而有待改进。该文用携带麦角硫酶质粒（pET28a）的转化体进行异源表达，得到麦角硫酶粗酶液（酶活为458.6 U/mL），以EGT标准品和食药用真菌为样本，对EGT含量的酶法快速检测体系进行优化，并将食药用真菌EGT含量的酶法快速检测结果和高效液相色谱检测结果进行相关性分析，验证酶法快速检测方法的实际可行性。结果表明，在100μL样品中加入麦角硫酶粗酶液100μL(45.86 U),311 nm下监测反应混合物吸光度，时间不超过20 min，可以快速且准确地检测出样本的EGT含量。更多还原

【Abstract】 In the microbial screening of medicinal fungi with high ergothioneine(EGT)production,the existing EGT assay method requires improvement due to its cumbersome steps and the high cost of reagents and equipment. Transformants harboring the ergothionase plasmid(pET28a)were employed for heterologous expression to yield a crude ergothionase solution(enzyme activity was 458.6 U/mL). The rapid detection system of the enzymatic method was optimized using EGT standards and medicinal fungi as samples,and a correlation analysis was conducted between the results of the enzymatic method-based rapid detection and those of highperformance liquid chromatography(HPLC)to validate the practical feasibility of the enzymatic method for rapid detection of medicinal fungi. The results indicated that when 100 μL of sample solution was mixed with100 μL of crude enzyme solution(45.86 U),and the absorbance of the reaction mixture was monitored at311 nm for more than 20 min,the EGT content across samples could be detected rapidly and accurately.更多还原