原生质体诱变筛选高产纽莫康定B_0的菌株

Production of pneumocandin B_0 strain by mutagenesis of protoplast

目的对产纽莫康定B_0的丝状真菌(Glarea lozoyensis)ATCC 74030进行原生质体制备、再生及常压室温等离子体诱变育种进行研究,选育高产纽莫康定B_0的菌株。方法采用单因素实验法研究了原生质体形成和再生的最佳条件,并将原生质体通过常压室温等离子体诱变。结果 Glarea lozoyensis原生质体形成和再生的最佳条件为:菌体培养9d,0.6mol/L的葡萄糖溶液作为渗透压稳定剂,采用2%Yatalase+3%溶壁酶+1%蜗牛酶组成的复合酶系,34℃条件下酶解3.5h。经过常压室温等离子体诱变,筛选出的原生质体诱变菌株G.lozoyensis Q1,其产量达1.13g/L,相对出发菌株产量(0.81g/L)提高了39.5%,且连续传代5代遗传稳定。结论常压室温等离子体诱变提高ATCC 74030发酵单位效果明显,实验结果表明突变株G.lozoyensis Q1具有潜在的生产应用价值。

Objective The conditions of protoplast formation, regeneration and atmospheric and room temperature plasma （ARTP） mutagenesis of Glarea lozoyensis ATCC 74030 strain were studied to breed pneumocandin B0 high yield mutants strain. Methods Protoplast preparation and regeneration condition were investigated by single factor design method. The protoplast was studied by ARTP breeding system. Results As a result, the optimal condition for protoplast preparation and regeneration of G. lozoyensis were showed as follows： 9 days of mycelium growth time, 0.6mol/L glucose was used as osmotic pressure stabilize, the content of yatalase, lysing enzymes and snailase was 20, 30 and 10g/L respectively, enzymolysis temperature was 34℃, and enzymolysis time was 3.5h. The pneumocandin B0 high yield mutants strain with good genetic stability was obtained, numbered G. lozoyensis Q1. Its pneumocandin B0 yield was up to 1.13g/L, which was 39.5% higher than that of the parent strain. Conclusion We successfully bred a high-yielding pneumocandin Bo producing strain by ARTP breeding system, the results suggested that G. lozoyensis Q 1 was a promising pneumocandin B0 producing strain.