%0 Journal Article %T 重组大肠杆菌全细胞转化马来酸高效合成富马酸 %A 房月芹 %A 周丽 %A 刘文茂 %A 周哲敏 %J 食品与生物技术学报 %D 2016 %R 10.3969/j.issn.1673-1689.2016.12.014 %X 重组Escherichia coli细胞催化马来酸合成富马酸时,细胞中富马酸酶将产物富马酸转化成苹果酸,转化率达15.5%,降低了富马酸的转化率和纯度。将E. coli BL21(DE3)基因组中fumA-fumC基因敲除,并高效表达马来酸顺反异构酶,重组菌BL21(DE3)△fumA-fumC/pET24a-maiA经发酵罐培养,可产生64 g/L菌体,马来酸顺反异构酶表达量达306 U/mL。按照60 g/L的发酵液:2 mol/L富马酸为1∶4的体积比配置反应液,37 ℃反应1 h,富马酸转化率高达98.4%,仅产生0.7%的苹果酸副产物。该结果为全细胞催化法合成富马酸的工业化奠定了基础。</br>Fumarate could be produced from maleate with whole cell of recombinant Escherichia coli. However,malate,with a yield of 15.5%,was generated as byproduct by the cellular fumarase,resulting in a decreased fumarate yield and purity. The chromosomal genes,fumA and fumC,in E. coli BL21(DE3) were inactivated,followed by over expression of the maleate cis-trans isomerate. After cultured in a bioreactor,the resulting strain BL21(DE3)△fumA-fumC/pET24a-maiA could generate 64 g/L dry cell weight and maleate cis-trans isomerate of 306 U/mL. With a volumetric ratio of fermentation broth(60 g/L dry cell weight):fumarate(2 mol/L)=1∶4,fumarate yield of 98.4% and malate yield of 0.7% were produced after incubation at 37 ℃ for 1 h. These results laid a foundation for industrial production of fumarate by whole-cell biocatalyst %K 富马酸 马来酸顺反异构酶 全细胞催化 苹果酸< %K /br> %K fumarate %K maleate cis-trans isomerate %K whole-cell biocatalysis %K malate %U http://spyswjs.cnjournals.com/spyswjs/ch/reader/view_abstract.aspx?file_no=201612014&flag=1