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含血红蛋白基因(VHb)的铜绿假单胞菌重组菌的构建及鼠李糖脂表达条件的研究

Construction of recombinant strain containing vitreoscilla hemoglobin gene(VHb) of Pseudomonas aeruginosa and its rhamnolipid expressing conditions

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【作者】 冯蕾谭之京胡鸢雷杨新平陈竞秦新政唐娴林忠平

【Author】 FENG Lei~1,TAN Zhi-jing~2,HU Yuan-lei~2,YANG Xin-ping~1,CHEN Jin~1, QIN Xin-zheng~1,TANG Xian~1,LIN Zhong-ping~2 (1.Microbiology Institute of Xinjiang Academy of Agricultural Sciences,Urumqi 830091; 2.The College of Life Science,Beijing University,Beijing 100871)

【机构】 新疆农业科学院微生物所北京大学生命科学学院

【摘要】 将RDR(ribonucleotide diphosphate reductase)启动子驱动下的透明颤菌(Vitreoscilla sp.)血红蛋白(Vitreoscilla hemoglobin,VHb)基因的表达载体pSETRDR-VHb转入铜绿假单胞(Pseudomonasaeruginosa)S301菌株中。并对其中阳性转化子AY26菌株进行了鼠李糖脂表达条件的研究。正交实验L9(43)优化培养基,最佳组分为:硫酸镁0.075%、硝酸钠0.5%、清油3 mL/L、酒石酸钠0.4%。在限碳培养条件下,转化子SY26鼠李糖脂产量达到12.9 g/L,比对照菌株S301(8.4 g/L)提高150%,5 L发酵罐放大实验验证,重组菌AY26的表面活性剂产量达到33.12 g/L。

【Abstract】 In order to construct the Tn5 shuttle plasmid containing vitreoscilla hemoglobin(VHb) with strong expressed promoter RDR in bacteria,the high level biosurfactants producing strain S301 transudated with VHb gene using the technique of conjugation and chromosome transposition was labelled,selected and identified,then a high level biosurfactant expressed transformants was obtained.By orthogonal test,the optimized substrate components were 0.075%of MgSO4’7H2O,0.5%of sodium nitrate,0.4%of sodium tartrate,3 mL/L of bean oil.The result showed that the yield of rhamnolipid produced by SY26 reached 12.9 g/L,higher than that by S301(8.4 g/L) and the yield of biosurfactant produced by AY26 in 5 L fermenter was as high as 33.12 g/L.

【基金】 国家自然科学基金项目(20666006);国家科技支撑项目(2009BADA4B04);新疆维吾尔自治区高技术研究发展计划项目(201010104);新疆乌鲁木齐市科技局项目(Y101210003)
  • 【文献出处】 工业微生物 ,Industrial Microbiology , 编辑部邮箱 ,2012年03期
  • 【分类号】TQ920.6;TQ423
  • 【被引频次】3
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