一株铜绿假单胞菌的筛选鉴定及其产鼠李糖脂条件的优化  

Screening and identification of a Pseudomonas aeruginosa strain and its rhamnolipid cultivative optimization

作　　者：杨佐毅[1] 崔佳豪 李平凡[2] 王玉洁[1] 谢甫玉 卓逢 YANG Zuoyi;CUI Jiahao;LI Pingfan;WANG Yujie;XIE Fuyu;ZHUO Feng(School of Environmental Science and Engineering,Guangdong University of Technology,Guangzhou Guangdong 510006;School of Food Engineering and Biotechnology,Guangdong Industry Polytechnic,Guangzhou Guangdong 510300)

机构地区：[1]广东工业大学环境科学与工程学院,广东广州510006 [2]广东轻工职业技术学院食品与生物技术学院,广东广州510300

出　　处：《环境污染与防治》2018年第2期128-132,共5页Environmental Pollution & Control

基　　金：国家自然科学基金资助项目(No.41603072);广东省科技计划项目(No.2014A020216038);广东工业大学青年基金资助项目(No.14QNZD006)

摘　　要：从重油污染的土壤中分离出菌株GTX1~GTX4,采用溴化十六烷三甲基铵(CTAB)蓝色凝胶培养基和排油圈法对菌株进行初筛,其中菌株GTX4排油圈直径最大且稳定,通过生理生化实验和16SrDNA鉴定,GTX4为铜绿假单胞菌。将GTX4接种于甘油培养基,在33℃、150r/min条件下培养96h后,经亚甲基蓝-氯仿法和薄层层析(TLC)测定产物为鼠李糖脂,蒽酮-硫酸法测定培养液中鼠李糖脂质量浓度为0.741g/L,培养液表面张力29.87mN/m。通过单因素实验确定GTX4培养的最佳碳源和氮源分别为调和油、NaNO_3,由正交实验得到GTX4产鼠李糖脂的优化条件为调和油45.0g/L,NaNO_3 4.0g/L,微量元素溶液3.5mL/L,菌液接种量0.35%(体积分数),在33℃、200r/min下摇床培养96h,培养液中鼠李糖脂质量浓度可达2.256g/L,液相表面张力降至27.39mN/m。In this study,the series strains named GTX1-GTX4 were isolated from the heavy-oil-contaminated soil.CTAB blue gel plate and oil displacement ring method were utilized to screen the strains at first,in which the oil displacement ring of GTX4 was the largest and the strain was stable.Afterward,the strain GTX4 was identified as Pseudomonas aeruginosa by the physiological-biochemical experiments and 16S rDNA sequence determination.When the strain GTX4 was inoculumed into the glycerol culture and cultivated on the condition of 33℃,150 r/min for 96 h,the rhamnolipid was qualitatively indentified by the methylene blue-chloroform method and the thin layer chromatography(TLC).The rhamnolipid of 0.741 g/L in the culture was determinated by the anthrone-sulfuric method and the surface tension was 29.87 mN/m.Blend oil and sodium nitrate were chosen as the best carbon source and nitrogen source for GTX4 by the single factor experiments.The results of orthogonal experiment revealed that the optimize conditions were as follows:sodium hydroxide 45.0 g/L,sodium nitrate 4.0 g/L,trace elements 3.5 mL/L,inoculation 0.35%(volume fraction).The concentration of rhamnolipid in the post-culture medium was 2.256 g/L after 96 h of cultivation at 33℃and 200 r/min,and the surface tension of the liquid was reduced to 27.39 mN/m.

关 键 词：铜绿假单胞菌 鼠李糖脂 表面张力 正交实验 

分 类 号：X172[环境科学与工程—环境科学] X53