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目的将透明颤菌血红蛋白基因(vgb)整合到那西肽产生菌—活跃链霉菌(Streptomyces actuo-sus)的染色体上进行表达,从而提高菌体对溶解氧的利用,提高那西肽的发酵产量。方法采用分子生物学方法构建了携带vgb的重组质粒pZV02,采用接合转移的方法将其导入那西肽产生菌—活跃链霉菌中,采用抗生素抗性标记筛选法获得了基因重组菌株。结果在低溶氧的发酵条件下,重组菌株的那西肽产量显著高于对照菌株。结论 vgb在活跃链霉菌中的表达,可以改善菌体对溶解氧的利用,在低溶氧的条件下可以提高发酵产物那西肽的产量。
OBJECTIVE: The purpose of this study was to integrate the Vitreoscilla hemoglobin gene (vgb) into the chromosome of nosiheptide producing bacterium Streptomyces actuo-sus, so as to improve the use of dissolved oxygen and increase the fermentation yield of nosiheptide . Methods The recombinant plasmid pZV02 carrying vgb was constructed by molecular biology method. The recombinant plasmid pZV02 was introduced into nosiheptide producing Streptomyces - Streptomyces by conjugate transfer method. The recombinant strain was obtained by antibiotic resistance marker screening. Results Under the condition of low DO fermentation, the production of nosiheptide from the recombinant strain was significantly higher than that of the control strain. Conclusion The expression of vgb in Streptomyces can improve the utilization of dissolved oxygen in the bacteria, and increase the production of nosiheptide in the fermentation under low DO condition.