高产辅酶Q10结构类似物抗性突变株的选育

以土壤杆菌(Agrobacteriumsp.)TLY-4为出发菌株,采用70%致死剂量的NTG进行诱变处理,通过筛选抗辅酶Q10结构类似物维生素K3突变株,定向选育到了两株辅酶Q10高产突变株,编号为R-122和R-015,其摇瓶发酵72h时的辅酶Q10产量分别为57.3 mg/L和59.9 mg/L,较出发菌株提高了35.7%和41.6%。通过连续传代实验,表明突变株高产辅酶Q10的遗传性状稳定。实验以有机溶剂DMF和吐温-80共同增溶的方法,解决了维生素K3在培养基中易析出的问题,并确定了平板培养基中维生素K3的最小抑菌浓度为0.15 mg/mL。

Breeding of the analogue-resistant strains for high production of coenzyme Q10

Agrobacterium sp.TLY-4 was treated by NTG with the death-rate of 70%. Two vitamin K3-resistant mutants R-122 and R-015 with high production of Co Q10 were obtained.The CoQ10 yield of R-122 and R-015 reached 57.3 mg/L and 59.9 mg/L in shaking flask for 72 hours.They were 35.7% and 41.6% higher than that of the original strain respectively.The mutants R-122 and R-015 showed high genetic stability in subculture experiments.The problem of the insolubility of vitamin K3 in the medium was solved by adding N,N-dimethylformamide(DMF) and Tween-80.And the minimal inhibitory concentration of vitamin K3 in ager plat was 0.15 mg/mL.