重组海藻糖合酶工程菌高密度发酵条件的研究

目的:通过对前期构建的海藻糖合酶基因工程菌进行高密度发酵的研究,获得了其高密度工艺条件。方法:采用摇瓶发酵和10L自控罐高密度发酵研究了培养基、pH、发酵方式对工程菌生长及目的蛋白表达的影响,并考察了工程菌中重组质粒的遗传稳定性。结果:海藻糖合酶基因工程菌高密度发酵的培养基为2YT+0.2%葡萄糖,最适pH为7.0,发酵方式为分批补料,通过10L自控罐高密度发酵最终得到的工程菌细胞密度达到了50.78g/L,酶活达到了3.197U/mL。所构建的重组质粒在宿主中得到了稳定遗传。结论:优化了海藻糖合酶基因工程菌高密度发酵的条件,为海藻糖规模化生产奠定了基础。 

High cell density fermentation condition of trehalose synthase genetic engineering bacteria

Object:To obtain the high dense fermentation procedure of trehalose synthase(Tres)genetic engineering bacteria previously construct.Methods:Studied the influence of medium,pH,ferment mode for incubation and induction on the growth of recombinant genetic engineering bacterial and expression of target protein in a 10L auto control biostat fermentator,and explored the genetic stability of recombinant plasmid.Result:The high-density culture medium of trehalose synthase genetic engineering bacteria was 2YT+0.2%glucose,the optimum pH was 7.0,the ferment mode was fed-batch.After the recombinant genetic engineering bacterial strain cultured in 10L high-density fermentation,the density of bacteria reached 50.78g/L,the enzyme activity reached 3.197U/mL.The constructed recombinant plasmid was inherited steadily in host bacteria.Conclusion:The high-density fermentation condition of trehalose synthase genetic engineering bacteria was optimized.It laid a foundation of large-scale production of trehalose.