天然产物研究与开发 ›› 2020, Vol. 32 ›› Issue (9): 1484-1490.doi: 10.16333/j.1001-6880.2020.9.005

• 研究论文 • 上一篇    下一篇

一株胞外多糖产生菌的筛选鉴定及其多糖成分研究

陆娟1，周慧1，王贵生1，余梅霞1，唐俊1,2*


  

  1. 1阜阳师范大学生物与食品工程学院；2环境激素与生殖发育安徽省重点实验室，阜阳 236037

  • 出版日期:2020-09-28 发布日期:2020-09-24
  • 基金资助:
    国家级大学生创新训练项目（201710371033）

Screening and identification of an exopolysaccharide-producing strain and the investigation of its polysaccharide

LU Juan1,ZHOU Hui1,WANG Gui-sheng1,YU Mei-xia1,TANG Jun1,2*#br#

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  1. 1School of Biotechnology and Food Engineering,Fuyang Normal University; 2Anhui Province Key Laboratory of Environmental Hormone and Reproduction,Fuyang Normal University,Fuyang 236037,China

  • Online:2020-09-28 Published:2020-09-24

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摘要：

以菌落周围有粘稠分泌物为初筛标准从土壤中筛选胞外多糖产生菌，苯酚-硫酸法测定初筛菌胞外多糖的产量；将胞外多糖产生菌LE-3进行形态学观察、BIOLOG分析与16S rDNA鉴定；提取菌株LE-3胞外粗多糖，该粗多糖经Sevage法除蛋白、透析和CM-琼脂糖凝胶FF、DEAE-琼脂糖凝胶FF、丙烯葡聚糖凝胶S-200HR柱层析进行纯化，傅里叶红外光谱法、薄层层析法和高效液相色谱法分析胞外多糖的单糖组成。结果表明：从土壤中筛出26株符合初筛标准的菌株，3株产糖量在5 g/L以上，其中菌株LE-3产量为5.29 g/L；根据形态学特征、BIOLOG和分子生物学分析，初步鉴定菌株LE-3为一株解淀粉芽孢杆菌亚种（Bacillus amyloliquefaciens subsp. amyloliquefaciens）；该菌株胞外粗多糖经Sevage法除蛋白、透析和三次柱层析纯化后，得到单一组分的LE-3胞外多糖；傅里叶红外光谱法、薄层层析法和高效液相色谱法分析确定该多糖为果聚糖。

关键词: 胞外多糖, 鉴定, 芽孢杆菌, 纯化, 果聚糖

Abstract:

Exopolysaccharide-producing bacteria were isolated from the soil according to the mucoid secretion around the colony,the yield of extracellular polysaccharide was determined by phenol-sulfuric acid method.The exopolysaccharide-producing strain LE-3 was identified based on morphological characteristics,Biolog analysis and 16S rDNA sequence analysis.The crude extracellular polysaccharide isolated from strain LE-3 was purified by Sevage method,dialysis and CM-Sepharose FF chromatography,DEAE-Sepharose FF chromatography and Sephacryl S-200HR chromatography.The monosaccharide composition of the polysaccharide was analyzed by FT-IR,TLC and HPLC.Results showed that among the 26 strains isolated from soil on the basis of the first screening standard,there were 3 strains that could produce exopolysaccharide more than 5 g/L,and the exopolysaccharide yield of strain LE-3 was 5.29 g/L.Strain LE-3 was identified as Bacillus amyloliquefaciens subsp. amyloliquefaciens based on morphological characteristics,Biolog analysis and 16S rDNA sequence analysis.A single component of exopolysaccharide isolated from strain LE-3 was obtained after deproteinization,dialysis and chromatography.Finally,the polysaccharide was determined to be a fructan by FT-IR,TLC and HPLC analysis.

Key words: exopolysaccharide, identification,  , Bacillus, purification, fructan

中图分类号:  Q539 Q93

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