Wang Qian, Liu Xiaolong, Qi Qingsheng
State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, People's Republic of China.
Appl Microbiol Biotechnol. 2014 May;98(9):3923-31. doi: 10.1007/s00253-013-5494-5. Epub 2014 Jan 15.
The biosynthesis of polyhydroxyalkanoate copolymers in Escherichia coli from unrelated carbon sources becomes attractive nowadays. We previously developed a poly(hydroxybutyrate-co-hydroxyvalerte) (PHBV) biosynthetic pathway from an unrelated carbon source via threonine metabolic route in E. coli (Chen et al., Appl Environ Microbiol 77:4886-4893, 2011). In our study, a citramalate pathway was introduced in recombinant E. coli by cloning a cimA gene from Leptospira interrogans. By blocking the pyruvate and the propionyl-CoA catabolism and replacing the β-ketothiolase gene, the PHBV with 11.5 mol% 3HV fraction was synthesized. Further, the combination of citramalate pathway with the threonine biosynthesis pathway improved the 3HV fraction in PHBV copolymer to 25.4 mol% in recombinant E. coli.
如今,利用大肠杆菌从非相关碳源生物合成聚羟基链烷酸酯共聚物变得颇具吸引力。我们之前通过大肠杆菌中苏氨酸代谢途径,从非相关碳源开发出了一条聚(3-羟基丁酸酯-co-3-羟基戊酸酯)(PHBV)生物合成途径(Chen等人,《应用与环境微生物学》77:4886 - 4893,2011年)。在我们的研究中,通过从问号钩端螺旋体克隆cimA基因,在重组大肠杆菌中引入了柠苹酸途径。通过阻断丙酮酸和丙酰辅酶A分解代谢并替换β-酮硫解酶基因,合成了具有11.5摩尔% 3HV组分的PHBV。此外,柠苹酸途径与苏氨酸生物合成途径的组合将重组大肠杆菌中PHBV共聚物的3HV组分提高到了25.4摩尔%。