摘要
为了筛选L-精氨酸的高产菌株,采用亚硝基胍对出发菌株ATCC14067(Brevibacterium flavum)进行诱变,结合抗精氨酸结构类似物D-精氨酸,S-甲基半胱氨酸平板抗性筛选高产菌株,并采用正交试验优化了种子培养基。结果显示,经过1 mg/m L的亚硝基胍诱变处理4 min后,采用14 mg/m L的D-精氨酸抗性平板和8 mg/m L的S-甲基半胱氨酸抗性平板筛选获得精氨酸高产菌株,由于解除精氨酸自身的反馈调节,L-精氨酸积累增大,产酸量达37.2 g/L,比野生菌株提高了128.2%,得到的高产菌株遗传性状稳定。通过对种子培养基进行正交试验优化,确定最终培养基配方为葡萄糖3.0%,玉米浆2.0%,硫酸铵2.0%,KH2PO40.10%,Mg SO4·7H2O 0.05%,尿素0.15%,在此发酵条件下,L-精氨酸产量达37.8 g/L。
In order to screen high-yielding strains ofL-Arginine （L-Arg）,starting strain ATCC14067（Brevibacterium flavum） was progressively mutagenized by nitrosoguanidine （NTG）.Anti-arginine structural analogs,D-arginine（D-Arg） and S-methyl cysteine （SMC） were used for high yield strain plate resistant screening.The seed medium was optimized by orthogonal experiment.The results showed that the high-yielding strains were selected by the treatment of 1 mg/ml NTG for 4 min and screened by arginine analog （14 mg/ml D-Arg and 8 mg/ml SMC） resistance plate,with the L-arginine accumulation increasing,the L-arginine production was 37.2 g/L.Compared to the wild strain,the production of L-arginine improved 128.2％,owing to the releasing of arginine feedback regulation,and the high-yielding strains had stable genetic traits.Through the optimization of seed medium by orthogonal experiment,result showed that the optimum formula was glucose 3.0％,com steep liquor 2.0％,（NH4）2SO4 2.0％,KH2PO4 0.10％,MgSO4·7H2O0.05％,urea 0.15％.Under this condition,the L-arginine production was 37.8 g/L.
出处
《中国酿造》
CAS
2014年第9期81-85,共5页
China Brewing
基金
广东高校特色调味品工程技术开发中心建设项目(GCZX-B1103)
关键词
L-精氨酸
诱变
平板抗性筛选
正交试验
L-arginine
mutagenesis
plate resistance screening
orthogonal experiment
