摘要
莽草酸是芳香族氨基酸合成中的重要中间代谢物,可用于多种药物的化学合成.在代谢工程理论的指导下,构建积累莽草酸的高产菌株.通过构建敲除编码莽草酸激酶的基因aroL与aroK的菌株阻断莽草酸的代谢;通过构建敲除编码奎尼酸/莽草酸脱氢酶的基因ydiB的菌株减少副产物的合成;同时通过构建人工操纵子过表达编码DAHP合酶、莽草酸脱氢酶、脱氢奎尼酸脱水酶和三脱氢奎尼酸合酶的基因aroG,aroE,aroD和aroB以强化莽草酸合成途径.最终得到E.coli SHIKΔaroLΔydiB菌株,摇瓶发酵能够积累莽草酸5.5g/L.
Shikimic acid is an important intermediate for aromatic amino acids.It can be used as building block for the chemical synthesis of various drugs.In this study,a high-yield shikimic acid producing strain was constructed through metabolic engineering.The genes aroLand aroK encoding the shikimate kinase were knocked out to block the shikimic acid catabolism and the gene ydiBencoding the quinic/shikimate dehydrogenase was deleted to reduce the synthesis of byproduct.The aroG,aroE,aroD and aroB genes encoding DAHP synthase,shikimic acid dehydrogenase, dehydroquinic acid dehydratase and dehydroquinic acid synthase were overexpressed with introduction of constructed operons,to strengthen the shikimic acid synthesis pathway.Finally,the obtained E.coli SHIKΔaroLΔydiBengineering strain resulted in a yield of shikimic acid of 5.5g/L by shake flask fermentation.
出处
《发酵科技通讯》
CAS
2017年第3期143-146,187,共5页
Bulletin of Fermentation Science and Technology
