红色素产生菌RZ21菌株的原生质体紫外诱变育种研究

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。

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

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.