1株降酚菌的原生质体制备和再生研究

Study on the Protoplast Preparation and Regeneration of a Strain of Phenol Degrading Bacteria

为了优化组合降酚菌原生质体制备与再生的条件。以从合肥钢厂焦化废水处理池的活性污泥中驯化筛选得到1株高效降酚假单胞菌为材料,通过正交实验,探讨蔗糖浓度、溶菌酶浓度E、DTA浓度和酶解时间对该菌原生质体制备与再生的影响。原生质体制备与再生的最优条件:蔗糖浓度20%,溶菌酶浓度0.5 mg/ml,EDTA浓度0.2%,酶解时间40 min。在此条件下,它的菌株形成率为75.3%,再生率为7.8%。影响降酚假单胞菌原生质体形成率的因素顺序为蔗糖浓度>溶菌酶浓度>作用时间>EDTA浓度。

The aim was to optimize the protoplast preparation and regeneration conditions of phenol degrading bacteria. With a high-efficiency phenol de-grading pseudomenad strain obtained through domestication and screening from active sewage in coking wastewater treatment pool of Hefei Steel Mill as ma-terials, the effects of 4 factors as sucrose concentration, lysozyme concentration, EDTA concentration and the time of enzyme hydrolysis on the protoplast preparation and regeneration of the bacteria was discussed through the orthogonal experiment. The optimum conditions of protoplast preparation and regener-ation were as follows： sucrose concentration of 20%, lysozyme concentration of 0.5 mg/ml,EDTA concentration of 0.2% and the time of enzyme hydroly-sis of 40 rain. Under these conditions, its strain formation rate was 75.3% and regeneration rate was 7.8%. The sequence of 4 factors influencing proto-olast formation rate of phenol degrading pseudomonad was sucrose concentration 〉 lysozyme concentration 〉 acting time 〉 EDTA concentration.