利用多功能等离子体诱变系统快速诱变选育γ-氨基丁酸高产菌株  

Rapid screening of a high yield γ-aminobutyric acid mutant by multifunction plasma mutagenesis system

作　　者：石秀峰[1] 纪曾春 常传友 马身喜 张变强 高强[1] 

机构地区：[1]天津科技大学生物工程学院,工业发酵微生物教育部重点实验室,天津300457

出　　处：《中国酿造》2016年第4期127-130,共4页China Brewing

基　　金：国家自然科学基金面上项目(31370075,31471725);国家重点基础研究发展计划‘973计划’(2013CB734004);国家高技术研究发展计划‘863计划’(2012AA021302)

摘　　要：为建立一种快速便捷的产γ-氨基丁酸(GABA)高产菌株的诱变选育方法,以产GABA的短乳杆菌(Lactobacillus brevis)TCCC 13007为出发菌株,采用多功能等离子体诱变系统(MPMS)对其进行诱变育种。最佳诱变条件为氮气流量10 L/min、照射间距3 mm、120 W功率条件下处理90 s,致死率>90%。将诱变后的菌悬液涂布到CaCO_3筛选平板上,以CaCO_3透明圈与菌落直径比值为依据,建立了平板-96深孔板-摇瓶的快速筛选体系。最终通过摇瓶复筛,选育得到了一株GABA产量明显提高的突变株L8,GABA产量达到(66.59±0.58)g/L,较出发菌株提高了11.41%。10次连续传代与摇瓶发酵实验表明,突变株L8的发酵性能和遗传稳定性良好。This work aims to establish an expeditious method for the screening of high-yield γ-aminobutyric acid （GABA） strains. A novel mutation approach, the multifunction plasma mutagenesis system （MPMS）, was employed to screening GABA-producing Lactobacillus brevis TCCC 13007. The optimal mutation conditions were N2 gas flow ratio 10 L/rain, irradiation distance 3 nun, power 120 W and radiation time 90 s. Under these conditions, the lethality ratio was over 90%. Based on the ratio of transparent circle to colony diameter on CaCO3 screening plate as the screening criteria, a ＂Petri dish-96 well plate-shake flask＂ screening protocol was established. The obtained mutant strain L8 was screened with high GABA production of （66.59±0.58） g/L, increased 11.41% than that of original strain. By a 10 th continuously subculture with shake flask GABA fermentation, the fermentation ability of the mutant strain L8 remained stable and showed high genetic stability.

关 键 词：多功能等离子体诱变系统 短乳杆菌 Γ-氨基丁酸 筛选 

分 类 号：TQ922[轻工技术与工程—发酵工程]