生物技术通报 ›› 2017, Vol. 33 ›› Issue (4): 191-197.doi: 10.13560/j.cnki.biotech.bull.1985.2017.04.025

• 研究报告 • 上一篇    下一篇

一株产鼠李糖脂菌株的筛选、鉴定及其产物特性分析

王江1, 2, 鞠酒2, 3, 曹海龙2, 郭卫华1, 尹恒2   

  1. 1. 大连海洋大学水产与生命学院,大连 116023;
    2. 中国科学院大连化学物理研究所天然产物与糖工程课题组 辽宁省碳水化合物重点实验室,大连 116023;
    3. 大连海洋大学食品科学与工程学院,大连 116023
  • 收稿日期:2016-10-28 出版日期:2017-04-25 发布日期:2017-04-25
  • 作者简介:王江,男,硕士研究生,研究方向:微生物和酶;E-mail:jiangwang1024@126.com
  • 基金资助:
    中科院青年创新促进会(2015144),大连市杰出青年科技人才(2015R010)

Screening and Identification of Rhamnolipid-producing Strains and Characterization of the Product

WANG Jiang1, 2, JU Jiu2, 3, CAO Hai-long2, GUO Wei-hua1, YIN Heng2   

  1. 1. College of Fisheries and Life Science,Dalian Ocean University,Dalian 116023;
    2. Natural Products and Glyco-Biotechnology,Dalian Institute of Chemical Physics,Chinese Academy of Sciences,Dalian 116023;
    3. College of Food Science and Engineering,Dalian Ocean University,Dalian 116023
  • Received:2016-10-28 Published:2017-04-25 Online:2017-04-25

摘要: 鼠李糖脂是最常见,研究最深入,应用最广泛的一类生物表面活性剂。从油田附近、沼气池旁的土壤中分离得到了25株菌,通过硫酸-苯酚反应,乳化实验,排油性实验,薄层色谱实验筛选产鼠李糖脂的菌株并表征产生的鼠李糖脂,通过16S rDNA 序列确定细菌的种属。硫酸-苯酚反应显示有2株菌可能产鼠李糖脂;5株菌的发酵液具有明显的乳化效果,命名为“其红”这株菌的乳化指数可达58.97%;1株菌的发酵液上清稀释10倍后排油圈直径仍可达3.53 cm;薄层色谱实验也显示“其红”菌产鼠李糖脂。在四个实验中 “其红”菌都检测到了鼠李糖脂,故确认“其红”菌可以产鼠李糖脂。16S rDNA序列分析表明“其红”菌属于希瓦氏菌属(Shewanella)并与腐败希瓦菌(S.putrefaciens LMG 26268(T))相似度最高。

关键词: 表面活性剂, 鼠李糖脂, 菌株筛选, 产物特性, 希瓦氏菌属

Abstract: Rhamnolipid is the most common biosurfactant that has been deepest studied and the most widely applied. Total 25 strains were isolated from soil of the near oil field and biogas plant. The strains that produced rhamnolipid were screened and characterized by sulfuric acid-phenol reaction,emulsification experiment,oil-spreading experiment,and thin layer chromatography experiment. The strains were identified by 16S rDNA sequence analysis. The sulfuric acid-phenol reaction showed that 2 strains might produce rhamnolipid;the fermentation supernatants of 5 strains showed emulsified effect and the emulsifying index of the strain was up to 58.97%. The oil-spreading diameter of the fermentation supernatant,diluted 10 times,of one strain reached 3.53 cm. The thin layer chromatography experiment verified that the strain “Qihong” did produce rhamnolipid. The rhamnolipid in the strain “Qihong” was detected by all four experiments,confirming that strain “Qihong” produced rhamnolipid. The 16S rDNA sequence analysis revealed that the strain “Qihong” belonged to Shewanella,and was the most similar to Shewanella putrefaciens LMG 26268(T).

Key words: surfactant, rhamnolipid, strain isolation, product characterization, Shewanella