发形霞水母共附生微生物的分离鉴定及其发酵液的抑菌活性研究  

Isolation and identification of the symbiotic and epiphytic microorganisms from the jellyfish Cyanea capillata and antibacterial activities of their fermentation broths

作　　者：尹慢慢 徐凤[2] 肖良[1] 王倩倩[1] 郑杰民[1] 张黎明[1] 

机构地区：[1]第二军医大学海医系海洋生物技术教研室,上海200433 [2]大连海洋大学食品科学与工程学院微生物教研室,辽宁大连116021

出　　处：《中国海洋药物》2014年第1期31-39,共9页Chinese Journal of Marine Drugs

基　　金：国家自然科学基金(41176126);国家高技术研究发展计划课题(2013AA092904)资助

摘　　要：目的对发形霞水母(Cyanea capillata)共附生微生物进行分离鉴定并测定其抑菌活性,以期获得具有较高抑菌活性的菌株。方法利用4种分离培养基从C.capillata各部位分离共附生微生物,首先对挑选的单个菌落进行反复的划线分离,获得各菌株的纯培养;利用形态学观察、生理生化特征检测和16SrDNA序列测定和分析等手段对获得的菌株进行鉴定;以金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌作为指示菌,利用滤纸片扩散法测定菌株发酵液的抑菌活性;通过正交试验方法进一步优化发酵条件,提高菌株发酵产生抑菌活性物质的能力。结果从C.capillata触手、胃囊、伞部、肌肉4个部位共分离出83株菌株,其中放线菌11株,细菌31株,真菌41株;其中菌株cmf-2经鉴定属于假单胞菌属。抑菌活性检测发现,在31株细菌中有14株对金黄色葡萄球菌、大肠杆菌、枯草芽孢杆菌均有较强的抑菌活性,占分离菌株的45.2%;正交试验结果显示,菌株cmf-2发酵液的最优条件为:培养温度35℃,pH 7.3,培养时间7d,蔗糖50g/L,蛋白胨10g/L。结论分离到的发形霞水母共附生微生物中含有较高比例的能产生抑制细菌生长的活性物质的细菌菌株,这些菌株具有较强的研究和开发价值。Objective To isolate and identify the symbiotic and epiphytic microorganisms from the jellyfish Cyanea capillata and find bacteria exhibiting high antibacterial activities from them. Methods Various culture media were utilized for isolation of the symbiotic and epiphytic microorganisms and strains selected were purified by repeated streaking. Microorganisms obtained were identified through morphological, physiological and biochemical characterization and 16SrDNA sequence analysis. Filter-paper disc diffusion method was used to evaluate the antibacterial activities of bacteria obtained, with Staphy- lococcus aureus, Escherichia coli and Bacillus subtilis being as the indicator bacteria. Orthogonal intersection experiment was used to optimize the prescription of the medium and the conditions of the fermentation broth to improve the antibacterial activity. Results 81 strains of symbiotic and epiphytic mi croorganisms were isolated from the jellyfish Cyanea capillata, including 11 stratus ot actmomyces, 31 strains of bacteria, 41 strains of fungi. 14 of the 31 strains of bacteria （45.2%） exhibited strong anti- bacterial activities against Staphylococcus aureus, Escherichia coli and Bacillus subtilis. Orthogonal intersection experiment results suggested that the best culture condition for cmf-2 fermentation broth was that 70 g/L of sucrose, 10 g/L of peptone, 1 L of distilled water,incubating at 37℃ for 6 days with pH 7.3. Conclusion The symbiotic and epiphytic microorganisms isolated from C. capillata contained high proportions of bacteria exhibiting strong antibacterial activities, which provided strains basis for further researches and exploitations.

关 键 词：发形霞水母 共附生微生物 分离鉴定 抑菌活性 

分 类 号：R1[医药卫生—公共卫生与预防医学]