原生质体融合构建高效降解工程菌的研究

Construction of High-Efficiency Degradation Engineering Microorganism from Protoplast Fusion

研究了球形红假单胞菌与酿酒酵母菌的原生质体融合条件 ,并对融合子性能进行了测定 .结果表明 :两种菌株分别在溶菌酶—— EDTA和蜗牛酶——巯基乙醇处理后形成原生质体 ,两种原生质体在聚乙二醇 ( PEG)和 Ca2 + 的诱导下发生融合 .最佳融合条件是 ,Ca2 + 浓度为 5 0 mmo L· L- 1 ,PEG质量分数为 3 0 % ,融合温度为 3 0℃ ,融合时间为 3 0 min,该条件下融合率为 6.8× 10 - 6 ;该融合子具有两亲株的特性 ,适应能力增强 ,比降解率是酵母的 1.88倍 .

The results of the protoplast fusion between Rhodopseudomonas spheroides and Sacchaaromyces cerevisiae were studied in this paper. Rhodopseudomonas spheroides cells were treated with lysozyme—EDTA system in order to obtain protoplasts, Sacchaaromyces cerevisiae cells were treated with snail enzyme—sulfhydryl alcohol system. Induced by polyethleneglycol(PEG) and calcium (Ca 2+ ),the fusion of the two protoplasts took place.The optimum conditions of the fusion were Ca 2+ concentration 50 mmol·L -1 ,PEG quality percentage 30%,fusion temperature 30℃, fusion time 30 min. The fusion frequency was 6.8×10 -6 under that condition. The fusant has the character of Rhodopseudomonas spheroides and Sacchaaromyces cerevisiae and a better adaptability in the process of the treatment.And the degradation ratio of the fusant is 1.88 times that of Saccharomyces cerevisiae.