重离子束12C6+累进辐照诱变柠檬酸菌株选育研究

应用大剂量重离子束12C6+ 对菌株H3001 进行二次辐照选育,对初选获得的高产柠檬酸菌株进行摇瓶发酵试验及10~ 100 L 中试发酵罐试验,采用酸碱中和法测定发酵液中柠檬酸的含量。结果表明:当二次重离子12C6+剂量为857.8 Gy 时,致死率和正突变率达到最大值,分别为94.5% 和8%。通过摇瓶发酵试验,最终获得一株高产柠檬酸菌株hw317,控制该菌株发酵周期为60 h,柠檬酸酸度能达到19.2±0.2%。Heavy 12C6+ ion beams in various high doses were employed to irradiate H3001 strain for screening Aspergillus niger strain for hyper citric acid production. Three high-yield strains were obtained after shaker fermentation test. Among the three strains, the strain hw317 was implemented shaker fermentation for stability test and 10~100 L pilot fermentation tank for citric acid productive maximization. Acid-base neutralization method was applied to determinate the content of citric acid in fermented liquid. The results showed that: when the secondary heavy ion 12C6+ dose was 857.8 Gy, both of the fatality rate (94.5%) and the positive mutation rate (8%) were highest. Through the shaker fermentation tests and 10 ~ 100 L pilot fermentation test, one strain hw317 was screened and obtained for hyper citric acid production. Consequently, the final citric acid acidity can reach up to 19.2±0.2% with controlling fermentation cycle for 60 h.