木质素降解菌L_1原生质体的形成和再生

Formation and regeneration from lignin degradation fungus L_1 protoplast

从自然界筛选出一株降解木质素高的白腐真菌 L1,对其原生质体的形成和再生进行了研究。在 OS培养基中生长的菌丝体 ,原生质体数量较高。在液体培养条件下 ,于 OS培养基中培养 60 h的菌丝体 ,用 0 .3% β-巯基乙醇与酶液同时处理菌丝体 ,采用 p H5 .0的混合酶 (蜗牛酶∶纤维素酶∶溶菌酶的最佳浓度比为 5∶ 4∶ 1 ) ;在 30℃酶解 4h;用 0 .4mol/L NH4Cl,1 0 mmol/L Mg SO4作渗透压稳定剂时 ,原生质体数量达到 4.32× 1 0 5个 /mg。

White rot fungus L 1 with high ability to degrade lignin was isolated from nature, and the formation and regeneration of L 1 protoplast was studied. By using the mycelia growing in the OS media, the higher yield of L 1 protoplast can be obtained . There was the highest yield of L 1 protoplast, up to 4.32×10 5/mg, when the mycelia growing in the OS media on the condition of liquid culture for 60 hours were dealt with 0.3% β mercaptoethanola and compound enzyme together by using the osmotic pressure stabilizer (0.4 mol/L NH 4Cl, 10 mmol/L MgSO 4) for 4 hours in 30℃. The compound enzyme was composed of snailase , celluase, lysozyme at the best concentration ratio of 5∶4∶1, pH value 5.0. The OS double layer regeneration media was the best media for L 1 protoplast regeneration.