摘要
三角酵母由于其胞内酶D-氨基酸氧化酶活力较高,具有一定的应用价值。关于这种酶的开发和利用国内外均有报道。本文报告利用原生质体融合法来进行三角酵母菌种选育,以期获得D-氨基酸氧化酶活力较高的菌株。
Three auxotrophic mutants of T. variabilis have been selected after mutation with NTG and LiCl: TV_(101), Met^-; TV_(102), Met^- V_(B6)^-; TV_(108), His^-. The regeneration efficiency of protoplasts was closely connected with the conditions of protoplast formation. The conditions used for protoplast formation were as follows: culture incubated for 32 hrs; Tris buffer 0.05mol/L, pHT.0; mercaptoethanol 0.25%; snail enzyme 1%; treated for 1.5hrs; 28—30℃. The conditions for regeneration were: pH 5.5—6.5, sucrose 0.6mol/L used as osmotic stabilizer. In complete medium added with 0.6mol/L sucrose, the percentage of regeneration amounted to 8.2%; while in minimum medium with 0.6mol/L sucrose, it was only 0.5%. 35% PEG6000 and 40% PEG4000 showed similar effects to protoplast fusion. The fusion frequency of the two recombinations TV_(101)×TV_(103) and TV_(102)×TV_(103) are 5.0×10^(-7) and 1.3×10^(-6) respectively.
出处
《微生物学报》
CAS
CSCD
北大核心
1991年第3期251-253,共3页
Acta Microbiologica Sinica
关键词
三角酵母
原生质体
Trigonopsis variabilis
Protoplast fusion
