基于抑菌活性的ε-聚赖氨酸的微孔板生物检测法  

A microplate bioassay to determine ε-polylysine based on antimicrobial activity

作　　者：骆健美[1,2,3] 成永新[3] 李培君[3] 王建锋[3] 郑宇[1,2,3] 王敏[1,2,3] 

机构地区：[1]天津科技大学工业发酵微生物教育部重点实验室,天津300457 [2]天津市工业微生物重点实验室,天津300457 [3]天津科技大学生物工程学院,天津300457

出　　处：《微生物学通报》2011年第7期976-981,共6页Microbiology China

基　　金：天津市高等学校科技发展基金计划项目(No.20080603);国际科学基金项目(No.F/4239-1)

摘　　要：基于ε-聚赖氨酸的抑菌活性,以96微孔板为平台,建立适合大规模样品快速分析的微孔板生物检测法。结果表明,藤黄微球菌(Micrococcus luteus)为最适敏感指示菌,当敏感指示菌初始浓度为107-108 CFU/mL,培养时间为4 h时,ε-聚赖氨酸浓度在100.00-500.00 mg/L范围内与抑菌率呈较显著的线性关系(R2=0.997 5)。该方法不仅表现出良好的精密度和准确度,与HPLC和甲基橙法的实验结果对比,微孔板生物检测法的相对标准偏差(RSD)和相对偏差(RD)分别小于3.5%和3.0%,说明该方法是一种与HPLC法有类似分析精度,但能直接反映样品的抑菌活性,且更适合大规模样品快速检测的方法,可用于发酵过程中ε-聚赖氨酸产量的检测和突变株的高通量筛选。Based on the antimicrobial activity of ε-polylysine,a 96-well microplate bioassay was estab-lished to rapidly determine large numbers of ε-polylysine samples.The results showed that a high cor-relation coefficient（R2=0.997 5） was obtained for linear regression in the ε-polylysine concentration range of 100.00-500.00 mg/L with 107-108 CFU/mL indicator strain（Micrococcus luteus） and the in-cubation time of 4 h.Using these parameters,the microplate bioassay has showed good precision and accuracy,and the relative standard derivation（RSD） and relative derivation（RD） were lower than 3.5% and 3.0% respectively.Compared with the results by Itzhaki method and HPLC assay,the microplate bioassay showed no considerable difference with HPLC,but could directly indicate the inhibition activ-ity and more suitable for rapid determination of numerous samples,which could be used for ε-polylysine quantification in fermentation process and high throughput screening from large numbers of mutants.

关 键 词：Ε-聚赖氨酸 微孔板 生物检测 抑菌活性 藤黄微球菌 

分 类 号：Q936[生物学—微生物学]