作　　者： ; ; ; ; ; ;

机构地区： 广西大学生命科学与技术学院发酵与酶工程研究所

出　　处： 《工业微生物》 2010年第3期39-43,共5页

摘　　要： 利用Red重组系统构建了大肠杆菌JM109甘油激酶基因(glpK)和甘油脱氢酶基因(gldA)缺失的双突变菌株JM109B,然后将表达酿酒酵母3-磷酸甘油脱氢酶基因(GPD1)和3-磷酸甘油酯酶基因(HOR2)的质粒pSE-gpd1-hor2转化到JM109B突变菌株中,在含1%葡萄糖的摇瓶发酵培养基中37℃发酵24 h,甘油的最高产量为5.61 g/L,是原始菌株JM109/pSE-gpd1-hor2甘油产量的1.59倍;在30 L发酵罐中发酵28 h,甘油的最高产量为103.12 g/L,是原始菌株JM109/pSE-gpd1-hor2甘油产量的1.59倍,是原始菌株BL21/pSE-gpd1-hor2甘油产量的1.41倍,葡萄糖转化率为50.39%。 By using Red recombination system, E. coli JM109 mutational strain with genes glpK and gldA disruption was constructed. The mutational strain was named E. coli JM109B. Then, the recombinant plasmid pSE-gpdl-hor2, which expressed 3-phosphate dehydrogenase and glycerol 3-phosphatase from Saccharornyces cerevisiae, was transformed into E. coli JM109B. And the recombinant microorganism was incubated in minimal media with 1% glucose shake-flasks at 37 12 with vigorous shaking for 24 h. Its maximal concentration of glycerol was 5.61 g/L, which was 1.59 times than that of E. coli recombinant strain JM109/ pSE-gpdl-hor2. The recombinant strain was fermented at 37 12 for 28 h. The maximal concentration of glycerol was 103.12 g/L, which was 1.59 times than that d E. coli JM109/pSE-gpdl- hor2 and was 1.41 times than that of E. coli BI21/pSE-gpdl-hor2, respectively. The conversion rate of glucose to glycerol was 50.39 %.

关 键 词： 甘油 基因敲除 重组系统 大肠杆菌突变菌株

领　　域： [生物学]