基因组改组快速提高谷氨酸棒杆菌L-鸟氨酸产量-食品科学.PDFVIP

206 2012, Vol. 33, No. 15 食品科学 ※生物工程 张媛媛 1,2 ，李家洲1 (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) ：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。 ：基因组改组；谷氨酸棒杆菌；L - 鸟氨酸；产量 Genome Shuffling for Rapid Improvement of L-Ornithine Production by Corynebacterium glutamicum ZHANG Yuan-yuan1,2 ，LI Jia-zhou1 (1. Department of Food and Bioengineering, Guangdong Industry Technical College, Guangzhou 510300, China； 2. School of Life Sciences, Sun Yat-sen University, Guangzhou 510300, China) Abstract ：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. Key words ：genome shuffling ；Corynebacterium glutamicum；L-ornithine ；production 中图分类号：Q939.97