周万龙,焦学,孙亚杰,等.一株海洋来源鞘氨醇单胞菌胞外多糖的含量测定[J].中国海洋药物,2017,36(2):27-34.
一株海洋来源鞘氨醇单胞菌胞外多糖的含量测定
Determination of extracellular polysaccharides produced by marine derived Sphingomonas sp. WG
投稿时间:2016-09-08  修订日期:2016-11-03
DOI:
中文关键词:  鞘氨醇单胞菌  胞外多糖  苯酚-硫酸法  条件优化
English Keywords:Sphingomonas sp. WG  extracellular polysaccharides  phenol-sulfuric acid method  process optimization
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作者单位E-mail
周万龙 中国石油大学(华东) 402356382@qq.com 
焦学 中国石油大学(华东) 生物工程与技术中心  
孙亚杰 中国石油大学(华东) 生物工程与技术中心  
李慧 中国石油大学(华东) 生物工程与技术中心  
朱虎* 中国石油大学(华东) 生物工程与技术中心 zhuhu@upc.edu.cn 
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中文摘要:
      目的 建立一种简便快捷的测定海洋来源鞘氨醇单胞菌分泌的胞外多糖威兰胶含量的方法。方法通过测定威兰胶同葡萄糖的换算因子以及发酵液中葡萄糖和总糖的浓度,建立了一种利用苯酚-硫酸法测定发酵液中威兰胶含量的方法,并采用单因素及正交实验对该方法的影响因素进行了优化及方法学验证。 结果 最佳的优化条件为:苯酚用量为0.8 mL、浓硫酸用量为6 mL、显色时间为30 min、显色温度为80 ℃。方法学验证结果表明,该方法的平均回收率为98.94%,相对标准偏差(RSD)为0.53%;显色产物在2 h内稳定,RSD为0.48%;仪器的精密度良好,RSD为0.77%。结论 建立的苯酚-硫酸法可用于发酵液中威兰胶的含量测定。
English Summary:
      Objective To establish a simple and rapid method for determination of the concentration of welan gum, extracellular polysaccharide produced by marine derived bacterium Sphingomonas sp. WG. Methods An phenol-sulfuric acid colorimetry method was developed to calculate the production of welan gum by the estimation of the conversion factor of welan gum to glucose, the concentration of residual glucose and total sugars in fermentation broth. Furthermore, the influence factor of the methods were optimized by single factor and orthogonal experiment, and validation of the method was carried out. Results The optimum determination conditions were 0.8 mL phenol, 6 mL sulfuric acid, reaction at 80°C for 30 min. The results showed that the average recovery rate is up to 98.94%, the RSD is 0.53%. The chromogenic product could keep stable within 2 h and RSD is 0.48%. In addition, the precision of the instrument is good, RSD is 0.77%. Conclusion The developed phenol sulfuric acid method can be used for quantitative determination of welan gum in fermentation broth.
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