基因组改组快速提高谷氨酸棒杆菌L-鸟氨酸产量  

Genome Shuffling for Rapid Improvement of L-Ornithine Production by Corynebacterium glutamicum

作　　者：张媛媛[1,2] 李家洲[1] 

机构地区：[1]广东轻工职业技术学院食品与生物工程系,广东广州510300 [2]中山大学生命科学学院,广东广州510300

出　　处：《食品科学》2012年第15期206-209,共4页Food Science

基　　金：国家自然科学基金面上项目(20876181)

摘　　要：以谷氨酸棒杆菌ATCC13032为出发菌,应用基因组改组技术快速提高L-鸟氨酸产量。经过紫外线、亚硝基胍和甲基磺酸乙酯分别诱变处理,获得6株产量有所提高的突变株,以此构建用于基因组改组的候选菌库。考察培养基成分对原生质体再生率的影响。经过两轮的灭活原生质体递推式融合,以磺胺胍和氟化钠为双抗性筛选标记,共筛选出2株遗传性能稳定的改组菌株。其中改组菌株F2-6摇瓶发酵72h,积累L-鸟氨酸产量为2.99g/L,是出发菌株的13.6倍。结果表明基因组改组技术能够在短期内使谷氨酸棒杆菌的L-鸟氨酸产量得以提高。Genome shuffling was applied to Corynebacterium glutamicum ATCC13032 to achieve rapid improvement of L-ornithine production.Six mutant strains with subtle improvements in L-ornithine production were obtained by treatment with UV light,nitrosoguanidine or ethyl methane sulfonate and subjected to recursive protoplast fusion.The effects of medium components on the protoplast regeneration rate were investigated.High-yield strains were screened using plates containing different concentrations of sulfaguanidine and NaF.After two rounds of genome shuffling,two shuffled strains producing L-ornithine at high level with good genetic stability were selected and named as F2-6 and F2-29.After 72h of shake-flask fermentation,the L-ornithine yield of strain F2-6 reached 2.99 g/L,which was 13.6 times higher than that of the original strain.

关 键 词：基因组改组 谷氨酸棒杆菌 L-鸟氨酸 产量 

分 类 号：Q939.97[生物学—微生物学]