摘要
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。
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 highdensity 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 largescale production of trehalose.
出处
《食品工业科技》
CAS
CSCD
北大核心
2012年第17期154-158,共5页
Science and Technology of Food Industry
基金
天津市应用基础及前沿技术研究计划(10JCYBJC09600
10JCYBJC05000)
国家自然科学基金(21076162)
关键词
海藻糖合酶
基因工程菌
高密度发酵
trehalose synthase（Tres）
genetic engineering bacteria
highdensity fermentation
