ε-聚赖氨酸产生菌的筛选及鉴定  

Screening and identification of ε-poly-lysine producing strain

作　　者：杜国丰[1] 辛俊宏 李学雷[1] 牟涛[1] 张林[1] 陈红漫[3] 刘凤翊 DU Guofeng;XIN Junhong;LI Xuelei;MU Tao;ZHANG Lin;CHEN Hongman;LIU Fengyi(Department of Chemical and Material Engineering,Yingkou Institute of Technology,Yingkou 115014,China;Educational Adminstrafion office of Yingkou Institute of Technology,Yingkou 115014,China;College of Biotechnology,Shenyang Agricultural University,Shenyang 110866,China)

机构地区：[1]营口理工学院化学与材料工程系,辽宁营口115014 [2]营口理工学院教务处,辽宁营口115014 [3]沈阳农业大学生物科学技术学院,沈阳110866

出　　处：《黑龙江畜牧兽医》2018年第13期20-24,28,240,241,共8页Heilongjiang Animal Science And veterinary Medicine

基　　金：辽宁省教育厅科学研究一般项目(L2014595)

摘　　要：为了筛选和鉴定ε-聚赖氨酸（ε-poly-lysine,ε-PL）产生菌,试验利用酸性品红从辽宁营口东部山区土壤中筛选到一株放线菌E301,采用薄层层析法分析发酵产物及其酸水解成分,水杨醛保护法分析产物酰胺键连接方式,红外光谱法对产物进行表征,通过形态观察、培养特征、生理生化特征并结合16S r DNA序列分析对菌种进行鉴定。结果表明：放线菌E301发酵产物与Dragendorff试剂、甲基橙试剂作用都有沉淀产生,薄层层析显示发酵产物的酸水解成分为单体赖氨酸,产物酰胺键连接方式为ε型,红外光谱图与标准ε-聚赖氨酸一致,形态学及分子生物学鉴定放线菌E301与白色链霉菌的同源性大于99%。说明放线菌E301为白色链霉菌Streptomyces albus。The aim of the present study was to screen and identify the ε-poly-lysine producing strain. An actinomycetes E301 was isolated from the soil sample collected at eastern mountainous area of Yingkou by using the acid fuchsin. The fermentation product and its acid hydrolytic components were analyzed by thin layer chromatography,and the connection mode of amide linkage was analyzed by salicylaldehyde protection method. The product was characterized by infrared spectroscopy and identified by morphological observation,culture characteristics,physiological and biochemical characteristics and 16 S r DNA sequence analysis. The results showed that the fermentation product of actinomycete E301 was precipitated with dragendorff reagent and methyl orange reagent,respectively. Thin layer chromatography showed that the acid hydrolysis product was monomeric lysine. The bonding mode of product amide linkage was epsilon type. The infrared spectrogram was consistent with the standardε-poly-lysine. Morphological and molecular biology analysis showed that the homology of actinomycete E301 and Streptomyces albus was more than 99%. The results indicated that the actinomycete E301 was Streptomyces albus.

关 键 词：Ε-聚赖氨酸 筛选 放线菌 薄层层析 红外光谱 白色链霉菌 

分 类 号：Q517[生物学—生物化学] Q93