ε-聚赖氨酸生产菌株的筛选和鉴定

Screening and Identification of ε-PL Producing Strain

建立了一种灵敏的ε-PL产生菌高通量的筛选方法。在分离培养基中加入150mg/LK2Cr2O7,以有利于放线菌的富集分离;并添加美蓝染料,利用产碱菌株碱性分泌物和美蓝之间的静电作用而形成独特的透明圈,从而灵敏地筛选得到产碱菌株;利用Dragendorff试剂与生物碱特有的颜色反应从产碱菌株中检出得到46株生物碱产生株;最后对产生物碱的菌株的发酵液进行ε-PL含量分析,确定4株ε-PL产生菌株。对其中一株ε-PL高产菌PL6-3菌株进行形态、生理生化和16SrDNA分析,结果表明PL6-3为北里孢菌属(Kita-satospora)。

A simple and sensitive method was developed to screen ε-PL produced strains from soils. 150mg/L K2Cr2O7 was added to the solid medium for actinomycetes enriching. A basic dye, methylene blue, incorporated in the agar pla.te to detect alkali preducers, because dye reacted with the secreted basic polymers by electrostatic interaction with the secreted basic pdymers and formed special zoon. And then dragendorff regent was used to detect alkaloid producers. Four ε-PL producers were obtained by TLC analysis. Meanwhile, phylogenetic analysis of PL6-3 strain, including morphology, physiological and biochemical characters and chemotaxomy were performed and phylogenetic tree was constructed based on the 16S rDNA sequences. And the results indicated that PL6-3 strain is a member of Kitasatospora.