产生物活性物质海洋放线菌的筛选和鉴定  

Screening and identification of active substances from Marine actinomycetes

作　　者：王艺颖[1] 丛丽娜[1] 王伟[1] 孙珊[1] 于咏梅[1] 孙野[1] 李秋月[1] 李成[1] 

机构地区：[1]大连工业大学生物工程学院,辽宁大连116034

出　　处：《工业微生物》2017年第3期30-35,共6页Industrial Microbiology

基　　金：辽宁省自然科学基金项目(201602047);国家海洋公益性行业科研专项项目(201405003);辽宁省教育厅重点实验室(LZ2014029)

摘　　要：本实验从辽宁大连海域的海水、海泥和海参养殖圈的样品中分离得到38株海洋放线菌,以金黄色葡萄球菌、溶壁微球菌、枯草芽孢杆菌、副溶血性弧菌和铜绿假单胞杆菌为指示菌,筛选出两株抑菌活性高的菌株,分别命名为HS-B31和HS-B34。16S r DNA测序鉴定及构建系统发育树的结果显示,它们都属于放线菌目、链霉菌属的不同种。对两株链霉菌的发酵上清液进行萃取、粗提和浓缩得到粗提物,即B31和B34;抗菌作用的结果显示,这两种粗提物的抗菌效果均为显著。经薄层层析分析,并利用制备型层析板对粗提物B31和B34进行活性物质的分离制备,共得到八个组分,即B31-1、B31-2、B31-3、B31-4、B31-5和B34-1、B34-2、B34-3。用滤纸片法对这些组分进行抗菌检测,结果显示组分B31-3和B34-3不仅对革兰氏阳性菌金黄色葡萄球菌有较好的抑菌作用,而且对海洋致病菌革兰氏阴性菌副溶血性弧菌也显示出较强的抑菌效果。研究为新型抗生素的研制和应用提供了微生物新资源。In this study, 38 marine actinomycetes were isolated from samples of sea water, sea mud and sea cucumber farming circles in Dalian sea area. Five indicators such as bacteria, Staphylococcus aureus, Micrococcus lysodeikticus, Bacillus subtilis, Vibrio parahaemolyticus and Pseudomonas aeruginosa were chosen for screening. And two strains named as HS-B31 and HS-B34 exhibited strong antibacterial activity. Both strains were identified by 16S rDNA sequence, Blast and phylogenetic analysis. The results indicated that the two marine actinomyeetes belonged to different Streptomyces species. The fermentation supernatants of the two strains were extracted by ethyl acetate. The crude extracts were concentrated to obtain the crude substances, named as B31 and B34. It was also found that both substances showed remarkable antibacterial activity. The active substances were isolated and analyzed by thin-layer chromatography. Five components （B31-1, B31-2, B31-3, B31-4 and B31-5） were isolated from B31, and three components （B34-1,B34-2, B34-3 ） were isolated from B34. Then all components of antibacterial activity were analyzed. The results revealed that the compounds B31-3 and B34-3 had stronger antibacterial activity against the gram-positive bacterium of V. parahaemolyticus and the gram-negative bacterium of S. aureus. These results would provide new microorganism resources for the development and application of antibiotics.

关 键 词：海洋放线菌 16S RDNA 鉴定 抗菌活性物质 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]