高产细菌素植物乳杆菌的诱变选育研究
Study on screeging of high-yield bacteriocin producing Lactobacillus plantarum stains induced by mutations
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摘要: 为获得细菌素高产菌株,以植物乳杆菌（Lactobacillus plantarum）JLA-9为出发菌株,对其进行亚硝基胍（NTG）诱变、常压室温等离子体（ARTP）诱变,以及基因组改组。结果表明,亚硝基胍诱变的最佳浓度为4 mg/m L,经筛选得到两株突变株N4-26、N4-27,其抑菌效价分别为2531.93、3057.32 IU/m L;常压室温等离子体诱变最佳时间为10 s,经筛选得到两株突变株ARTP10-37、ARTP10-61,其抑菌效价分别为2974.27、3261.62 IU/m L。将上述抑菌效价提高的菌株经四轮基因组改组后,得到一株突变株F4-2,其抑菌效价达到7374.76 IU/m L,相对于原始菌株提高了2.35倍,且遗传稳定性良好。研究表明理化诱变结合基因组改组的方式是快速获得理想菌株的有效方法。Abstract: In order to obtain a high- yield bacteriocin producing strain,Lactobacillus plantarum JLA-9 was used as the starting stain and mutagenized with nitrosoguanidine( NTG),atmospheric and room temperature plasma( ARTP) and genome shuffling.The results showed that the optimal concentration of NTG was 4 mg / m L,and two mutants named N4- 26,N4- 27 were obtained,the antibacterial titer of which were 2531.93 IU / m L,3057.32 IU / m L respectively. The optimal irradiation time of ARTP was 10 s,and two mutants named ARTP10- 37,ARTP10- 61 were obtained,the antibacterial titer of which were2974.27 IU / m L,3261.62 IU / m L respectively. The four strains with higher inhibitory activity were subjected to recursive protoplast fusion.After four rounds of genome shuffling,a strain named F4-2 was screened.The antibacterial titer of strain F4-2was increased to 7374.76 IU / m L,2.35 times compared with the wild- type strain.Research showed that physical and chemical mutagenesis combined with genome shuffling was an effective method for obtaining ideal strains.
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