海洋来源微生物的分离培养与抗肿瘤活性筛选

目的 从海洋环境样品中分离纯化微生物，经发酵培养与活性筛选，获取抗肿瘤活性菌株以供筛选药源活性产物，获无活性菌株以供核糖体工程转化研究。方法 通过单菌落挑选与划线培养分离纯化微生物菌株，经摇床发酵和提取操作制备活性测试样品。采用MTT法结合显微镜下细胞形态学检测的方法，测试样品的抗肿瘤活性。结果 从渤海湾驴驹河潮间带海泥样品中分离得到了微生物127株，其中真菌80株、放线菌47株。在127株中100 mg·L^-1样品浓度下对K562细胞的抑制率大于40%的活性菌株为8株，占菌株总数的6.3%(其中放线菌4株，占放线菌数的8.5%，真菌4株，占真菌数的5%)，抑制率在20%～40%的放线菌6株，占放线菌数的12.7%。结论 从渤海湾海泥样品中分离得到真菌80株、放线菌47株，从中获得抗肿瘤活性真菌4株、放线菌10株，从放线菌中获得抗肿瘤活性菌株的频率远远高于真菌。活性菌株为寻找药源活性产物提供了菌株，无活性菌株则为核糖体工程拓展药源菌株来源研究提供了资源。

Isolation of marine-derived microorganisms and screening for their anti-tumor activity

Objective To isolate marine-derived microbial strains for obtaining the bioactive metabolite producing strains to investigate anti-tumor active metabolites and non-producing strains to provide the strain resource for ribosome-engineering study. Methods Microbial strains were isolated by single colony isolation technique. The anti-tumor activity was assayed by MTT method using K562 cells together with morphological observation of K562 cells under inversed light microscope.Results Total 127 microbial strains, including 80 fungi and 47 actinomycetes, were isolated from soil samples collected at the tideland of Bohai Bay around Lüjühe in Tanggu District of Tianjin, China. Among them, samples from 4 actinomycetic and 4 fungal strains inhibited the proliferation of K562 cells with the inhibition rates over 40% at the 100 mg·L^-1 of concentration, while the samples from another 6 actinomycetic strains showed anti-tumor activity with inhibition rates ranging in 20% to 40% at 100 mg·L^-1. Conclusion The 80 fungal and 47 actinomycetic strains were isolated from the Bohai marine-environment samples, from which, the 4 fungal and 10 actinomycetic strains that produce metabolites with anti-tumor activity have been obtained. The frequency of obtaining the bioactive metabolite producing strains is obviously higher in actinomycetes than in fungi. These microbial strains including both the bioactive metabolite producing and non-producing strains have provided microbial strain resources for further studies on the bioactive metabolites and on the alteration of metabolic capacity of microbes to produce bioactive metabolites by ribosome-engineering, respectively.