极地研究

极地研究 ›› 2014, Vol. 26 ›› Issue (3): 292-299.DOI: 10.13679/j.jdyj.2014.2.292

• 研究论文 • 上一篇    下一篇

海洋放线菌Y18的分子鉴定、生物活性及其抗菌物质发酵条件优化

苑孟1,2 俞勇2 李会荣2 董宁1,2 张晓华1   

  1. 1中国海洋大学海洋生命学院，山东 青岛266003； 2国家海洋局极地科学重点实验室，中国极地研究中心，上海 200136
  • 收稿日期:2013-09-04 修回日期:2013-10-22 出版日期:2014-09-30 发布日期:2014-09-30
  • 通讯作者: 俞勇
  • 基金资助:

    国家自然科学基金项目（冰雷达与卫星测高协同的南极冰下湖遥感识别研究）;上海市自然科学基金项目

MOLECULAR IDENTIFICATION, ANTIMICROBIAL ACTIVITY AND FERMENTATION CONDITIONS OF ANTIBIOTIC SUBSTANCES OF MARINE ACTINOBACTERIA NOCARDIOPSIS SP. Y18

YUAN Meng1,2, YU Yong2, LI Huirong2, DONG Ning1,2, ZHANG Xiaohua1   

  1.  
    1Department of Marine Biology, Ocean University of China, Qingdao 266003, China;
    2SOA Key Laboratory for Polar Science, Polar Research Institute of China, Shanghai 200136, China
  • Received:2013-09-04 Revised:2013-10-22 Online:2014-09-30 Published:2014-09-30

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摘要： 从北冰洋楚科奇海海洋沉积物中分离到一株放线菌Y18，其良好生长需要海水。经16S rRNA序列分析确定该菌株属于拟诺卡氏菌属（Nocardiopsis），次级代谢产物合成基因及抗菌活性筛选发现，它含有聚酮类Ⅱ型（PKSⅡ）和非核糖体含硫多肽类（non ribosome-bound sulfur-containing peptides，NRPS）化合物合成基因，并且对枯草芽孢杆菌和白色念珠菌有抑制作用。以枯草芽孢杆菌为指示菌对Y18的发酵条件进行了初步优化，适合的发酵条件为：发酵培养基（蛋白胨 2g，酪素水解物 2g，酵母膏 2g，葡萄糖 1g，海水 1L），种子液接种量4%，发酵温度28℃，发酵培养基初始pH值8.0，培养时间6天，装液量40%。并对Y18发酵液稳定性进行了测定，结果显示：发酵液对酸碱、热的稳定性都较强，4℃下贮藏发酵液的抗菌活性保留时间较长。

关键词: 海洋放线菌, 分子鉴定, 抗菌活性, 发酵条件, 北冰洋

Abstract: The actinomycete strain Y18 was isolated from ocean sediments collected from the Chukchi Sea in the Arctic Ocean. This actinomycete requires seawater for growth. Based on 16S rRNA sequence analysis, it belongs to the genus Nocardiopsis. Results of secondary-metabolite biosynthesis screening show that it contains polyethylene ketone type Ⅱ (PKSⅡ) and non ribosome-bound sulfur-containing peptide (NRPS) genes. It has antimicrobial activity against both Bacillus subtilis and Candida albicans. The optimal fermentation conditions of Y18 against Bacillus subtilis are as follows: fermentation medium containing peptone 2 g, casein hydrolysate 2 g, yeast extract 2 g, glucose 1 g, sea water 1 L, and seed liquid inoculum to 4%, an initial pH 8.0, a culture temperature of 28°C, a liquid volume of 20% in a 1 000 mL flask, and a fermentation time of 6 days. We found that antimicrobial activity remained stable with changes in heat, acid, alkali, and during storage at 4°C.

Key words: Marine actinomycetes, molecular identification, antibacterial activity, fermentation conditions, Arctic Ocean