重组大肠杆菌利用蔗糖及糖蜜发酵生产丁二酸  

Succinic acid production from sucrose and sugarcane molasses by metabolically engineered Escherichia coli

作　　者：李凤[1] 马江锋[1] 吴明科[1] 冀亚亮 陈吴方 任心怡[1] 姜岷[1] 

机构地区：[1]南京工业大学生物与制药工程学院材料化学工程国家重点实验室,江苏南京211816

出　　处：《生物工程学报》2015年第4期534-541,共8页Chinese Journal of Biotechnology

基　　金：国家重点基础研究发展计划(973计划)(No.2013CB733901);国家高技术研究发展计划(863计划)(No.2011AA02A203);江苏高校优势学科建设工程项目;新世纪优秀人才支持计划资助~~

摘　　要：富含蔗糖的甘蔗糖蜜可作为制备丁二酸的廉价原料。然而生产丁二酸的潜力菌株大肠杆菌Escherichia coli AFP111不能代谢蔗糖。为了使其具有蔗糖代谢能力,将E.coli W中非PTS蔗糖利用系统蔗糖通透酶的编码基因csc B,果糖激酶的编码基因csc K和蔗糖水解酶的编码基因csc A克隆并表达到AFP111中,获得重组菌株AFP111/p MD19T-csc BKA。经厌氧发酵验证,重组菌株72 h消耗20 g/L蔗糖,丁二酸产量达到12 g/L。在3L发酵罐中采用有氧阶段培养菌体、厌氧阶段发酵的两阶段发酵方式,厌氧发酵30 h,重组菌株以蔗糖和糖蜜为碳源丁二酸产量分别为34 g/L和30 g/L。结果表明,通过外源引入非PTS蔗糖利用系统,重组菌株具有较强的代谢蔗糖生长及合成丁二酸的能力,并且能够利用廉价糖蜜发酵制备丁二酸。Sugarcane molasses containing large amounts of sucrose is an economical substrate for succinic acid production. However, Escherichia coli AFP111 cannot metabolize sucrose although it is a promising candidate for succinic acid production. To achieve sucrose utilizing ability, we cloned and expressed csc BKA genes encoding sucrose permease, fructokinase and invertase of non-PTS sucrose-utilization system from E. coli W in E. coli AFP111 to generate a recombinant strain AFP111/p MD19T-csc BKA. After 72 h of anaerobic fermentation of the recombinant in serum bottles, 20 g/L sucrose was consumed and 12 g/L succinic acid was produced. During dual-phase fermentation comprised of initial aerobic growth phase followed by anaerobic fermentation phase, the concentration of succinic acid from sucrose and sugarcane molasses was 34 g/L and 30 g/L, respectively, at 30 h of anaerobic phase in a 3 L fermentor. The results show that the introduction of non-PTS sucrose-utilization system has sucrose-metabolizing capability for cell growth and succinic acid production, and can use cheap sugarcane molasses to produce succinic acid.

关 键 词：大肠杆菌AFP111 非PTS蔗糖利用系统 蔗糖 糖蜜 丁二酸 

分 类 号：TQ921[轻工技术与工程—发酵工程]