L-精氨酸产生菌诱变育种及发酵条件优化  

Breeding of L-arginine-producing Strain and Optimization of Fermentation Conditions

作　　者：周栋[1,2] 唐志如[1] 冯泽猛[1,2] 唐香山[1,2] 李丽立[1] 

机构地区：[1]中国科学院亚热带农业生态研究所亚热带农业生态过程重点实验室,湖南长沙410125 [2]中国科学院研究生院,北京100039

出　　处：《农业现代化研究》2010年第6期733-737,共5页Research of Agricultural Modernization

基　　金：中国科学院院长奖获得者科研启动专项资金"L-精氨酸高产菌种的研发及其应用"(编号:0723022110);国家自然科学基金面上项目"重组牛乳铁蛋白素在毕赤酵母中的表达及其作用机理研究"(编号:30700581);中国科学院亚热带农业生态研究所青年人才领域前沿项目"抗菌肽buforinII在毕赤酵母中的表达及其作用机理研究"(编号I:SACX-LYQY-QN-0701)

摘　　要：本文以Corynebacterium glutamicum DSM1412为出发菌株,经紫外线和1-甲基-3-硝基-1-亚硝基胍逐级诱变处理和选育,并采用N-硝基-L-精氨酸甲酯进行抗性选育,获得一株高产L-精氨酸和遗传性稳定的菌株CorynebacteriumglutamicumZDNTG5。该菌种产L-精氨酸的量可达31g/L,比出发菌提高了1.4倍。通过对发酵培养基优化,确定最佳摇瓶发酵条件为葡萄糖9.0%,玉米浆3.0%,(NH4)2SO46.0%,KH2PO40.05%,MgSO·47H2O0.03%,碳酸钙2.0%,接种量6%,pH7.0,装液量15mL/100ml,转速100rpm,30℃发酵培养96h。在此条件下发酵,产酸量达44g/L,产精氨酸的量比未经优化时提高了41.9%。A L-arginine-producing mutant Corynebacterium glutamicum ZDNTG5was selected out by arginine-methylester resistance after step-by-step mutagenic treatment with ultraviolet and N-methyl-N-nitro-N-nitrosoguanidine from Corynebacterium glutamicum DSM1412. This mutant has good stability of descendiblity of L-arginine and products 31 g/L L-arginine. After the optimization of fermentation medium by orthogonal analysis, results showed optimal medium containing of glucose 9.0%, Corn steep liquor 3.0%, （NH4）2SO4 6.0%, KH2PO4 0.05%, MgSO4·7H2O 0.03%, calcium carbonate 2.0%, inoculum size 6%, pH 7.0, liquid volume 15ml/100ml. Mutant ZDNTG5 could accumulate 44 g/L L-arginine on average, which increased about 41.9% more than before in this optimized medium when it was cultured at 30℃ for 4 days.

关 键 词：L-精氨酸 谷氨酸棒杆菌 发酵 诱变育种 

分 类 号：Q517[生物学—生物化学] Q815