铜绿假单胞菌高产鼠李糖脂菌株的筛选

doi:10.13560/j.cnki.biotech.bull.1985.2015.10.028

摘要/Abstract

摘要： 从多种来源筛选高产鼠李糖脂的菌株, 并研究菌种发酵特性和鼠李糖脂产物的理化性质。采用CTAB平板初步筛选鼠李糖脂合成菌株, 通过分析菌株的16S rRNA基因序列确定细菌种属, 采用薄层色谱、红外光谱分析产物性质。结果显示, 利用CTAB平板初筛获得163株阳性菌株, 初步发酵确定10株高产细菌鼠李糖脂的产量为12.2-17.7 g/L, 10株细菌均鉴定为铜绿假单胞菌。挑选产量最高的菌株B12, 分别以甘油、菜籽油、花生饼粉或葵花籽饼粉为碳源进行发酵, 发现菜籽油为合成鼠李糖脂的最佳碳源。进一步对比在35℃、37℃和40℃的发酵水平, 发现37℃条件下鼠李糖脂产量最高, 为26.8 g/L。最后, 对鼠李糖脂发酵产物进行了初步纯化, 并进行了薄层色谱和红外光谱分析。菌株B12能够合成较高水平的鼠李糖脂, 可能成为工业生产的候选菌株。

关键词: 生物表面活性剂, 铜绿假单胞菌, 鼠李糖脂, 条件优化

Abstract: This work aims to screen strains highly producing rhamnolipid from multiple sources, analyze the characterization of fermentation and physicochemical characteristics of rhamnolipid. CTAB（cetyltrimethyl ammonium bromide）methylene blue plate was used for primary screening the strains synthesizing the rhamnolipid. Then the strains were identified by analyzing 16S rRNA sequences, and the property of rhamnolipid was analyzed by TLC（thin layer chromatography）and FTIR（Fourier transform infrared spectroscopy）. Total 163 strains with a dark blue halo around the colony were selected for further analysis of producing rhamnolipid. Among them, 10 strains producing 12.2-17.7 g/L rhamnolipid were identified as Pseudomonas aeruginosa. Moreover, strain B12 yielding highest rhamnolipid was selected and used for the optimization of carbon resource, including glycerol, rapeseed oil, peanut cake or sunflower seed cake, and rapeseed oil was recognized as the optimal carbon source for the synthesis of rhamnolipid. Fermentation temperature was also evaluated at 35℃, 37℃ and 40℃, so the production of rhamnolipid was the highest of 26.8 g/L at 37℃. In addition, rhamnolipid produced by strain B12 was purified and analyzed by TLC and FTIR. In conclusion, strain B12 could synthesize a relatively high level of rhamnolipid and could be a candidate strain for industrial production.

Key words: bio-surfactant, Pseudomonas aeruginosa, rhamnolipid, optimization

张翠坤,常冬妹,杨洪江. 铜绿假单胞菌高产鼠李糖脂菌株的筛选[J]. 生物技术通报, 2015, 31(10): 177-183.

Zhang Cuikun, Chang Dongmei, Yang Hongjiang. Screening of Pseudomonas aeruginosa Strains Highly Producing Rhamnolipid[J]. Biotechnology Bulletin, 2015, 31(10): 177-183.

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