摘要
对采自海南、湛江等海域的海绵样品进行放线菌选择性分离，采用其发酵液进行抗肿瘤活性筛选，并对活性较好的菌株进行鉴定。采用MTT法筛选分离出的菌株，通过对菌株的培养特徵、形态特徵、生理生化特徵、16SrDNA序列测定及其系统发育分析对菌株HA01041进行鉴定。以含50μg/ml的重铬酸钾作抑制剂的海水高氏一号合成培养基，用于海绵放线菌的分离培养具有很好的选择性，从海绵样品中共分离得到放线菌165株，活性筛选得到细胞毒活性达80％以上的阳性菌株10株，其中菌株HA01041的发酵液细胞毒活性为91％。结合形态观察，生理生化特徵和16SrDNA序列的比封分析，初步鉴定菌株kHA01041为春季链霉菌Streptomyces verne。菌株HA01041的发酵液细胞毒活性为91％，初步鉴定菌株HA01041为春季链霉菌Streptomyces verne。
Objective To isolate actinomycete from marine sponge collected from the sea area in Hainan and Zhanjiang, screen their antitumor activity, identify strains with better antitumor activity. Methods The antitumor activity was assayed by MTI＂ method, the strain HA01041 was identified by culture characteristics, morphological characteristics, physiological and biochemical properties, 16S rDNA sequence and phylogenetic analysis. Results The results Gause 1 medium added 50~tg/mL K2Cr205 possessed good selectivity. 165 strains of actinomycete were isolated from sponge and among them 10 strains showed strong antitumor activity, and one actinomycete HA01041 possessed cytotoxic activity 91%. The mycelium morphology, physiological and biochemical properties, 16S rDNA and phylogenetic tree suggested that strain HA01041 is Streptomyces verne. Conclusions One actinomycete HA01041 possessed cytotoxic activity 91%, and strain HA01041 is Streptomyces verne.
