红色素产生菌RZ21菌株的原生质体紫外诱变育种研究  

Study on the UV Mutagenesis Breeding of Protoplast in Prodigiosin ProduDing Bacteria RZ21

作　　者：夏顺翔[1] 张魁[1] 贾秉晟[1] 杨启银[1] 薛建[1] 

机构地区：[1]南京师范大学生命科学学院南京微生物工程技术研究中心,江苏南京210046

出　　处：《安徽农业科学》2010年第6期2780-2782,2792,共4页Journal of Anhui Agricultural Sciences

基　　金：江苏省高新技术项目(BG2005326)

摘　　要：[目的]对红色素生产菌RZ21菌株的原生质体进行紫外诱变育种,以提高其红色素的产量。[方法]以粘质沙雷氏菌为供试菌种,研究其原生质的制备、再生的条件和该菌的原生质紫外诱变对红色素产量的影响。[结果]RZ21菌株以溶菌酶8 mg/ml,37℃作用30 min后,再加0.01 mol/L(终浓度)的EDTA作用20 min,原生质体形成率达81.9%,再生率达31.5%。该菌原生质体经紫外诱变原生质体处理及抗性平板、摇瓶两步筛选后,得到1株高产株RZ21-3,灵菌红素产量达到了0.813 g/L,比原始菌株提高了24.9%。[结论]溶菌酶8 mg/ml,37℃作用30 min后,再加0.01 mol/L(终浓度)的EDTA作用20 min为该菌原生体制备的最佳工作条件。[Objective]The aim is make UV mutagenesis breeding of the protoplast of haematochrome producing bacteria RZ21,in order to increase the output of prodigiosin.[Method]With S.marcescens RZ-21 as the tested strain,the conditions of producing,regenerating prodigiosin and the effect of UV inducement on the prodigiosin were studied.[Result]The production rate of protoplast was 81.9% and regeneration rate was 31.5%,under the conditions of interacting RZ-21 with lysozyme（8 mg/ml,37 ℃） for 30 min,then interacting it with EDTA（0.01 mol/L） for 20 min.The mutant with high production of prodigiosin,RZ21-3,was screened by UV mutagenesis,streptomycin-resistant flat-panel screening and re-selected by shaking fermentation.The production of prodigiosin of RZ21-3 in flask fermentation was 0.813 g/L,24.9% higher than the parent strain.[Conclusion]The optimized method of producing and regenerating prodigiosin of RZ-21 is interacting RZ-21 with lysozyme（8 mg/ml,37 ℃） for 30 min,then interacting it with EDTA（0.01 mol/L） for 20 min.

关 键 词：原生质体 紫外诱变 灵菌红素 发酵 

分 类 号：S182[农业科学—农业基础科学]