摘要
目的系统研究 6 ,2 2 二烯 5 ,8 过氧麦角甾 3 醇 (EP)产生菌粘帚霉属真菌F菌的原生质体制备、分离与再生的条件。方法通过研究不同的酶解系统、酶解时间、再生培养基等多种因素 ,考察它们对该菌原生质体得率及再生率的影响 ;同时 ,采用高效液相色谱法测定原生质体再生菌株中EP化合物的含量。结果F菌菌龄为 6 0h时 ,以 0 .5mol/L的甘露醇配制成 2 %的蜗牛酶和纤维素酶混合溶液 ,2 8℃酶解 4h ,原生质体得量较高。而以0 .5mol/L的甘露醇为稳渗剂的再生培养基时 ,该菌的原生质体再生率较高。结论从代谢产物的角度对该菌原生质体制备和再生方法的可行性进行分析 ,并探讨此过程对F菌生物合成EP的影响 ,为进一步对该菌的分子遗传及高产菌株的选育创造条件。
PurposeThe aim is to study the conditions of preparation and regeneration of Gliocladium sp. (F) protoplast. Methods Different enzyme systems, enzymolysis time, osmotic pressure stabilizers were studied to investigate their influence on the productivity and regenerating rate of Gliocladium sp. F protoplasts. The HPLC method was used to determine EP (6,22 diene 5,8 epidioxy ergosta 3 hydroxy) content.ResultsThe higher productivity of protoplasts was obtained when mycelia of strain F growing for 60 hours was digested at 28℃ for 4 hours by solution containing 2% cellulase and 2% helicase dissolved in 0.5 mol/L mannitol and the medium containing 0.5mol/L mannitol as osmotic pressure stabilizer would be suitable for protoplast regeneration. According to the EP productivity detected by HPLC, high positive rate of the regenerated strains growing in the medium containing 0.5mol/L mannitol could be got. ConclusionThe results will promote the research of strain F mutantgenesis and will be helpful for obtaining the strain more effectively biosythesizing compound EP.
出处
《中国生化药物杂志》
CAS
CSCD
2001年第2期67-70,共4页
Chinese Journal of Biochemical Pharmaceutics
基金
国家自然科学基金!项目 (39970 0 18)
