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(Pseudomonas aeruginosa)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3mL/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％,5L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.%In order to construct the Tn5 shuttle plasmid containing vitreosdlla 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.
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