产纤维素酶原生质体形成和再生条件选择  

Optimized Conditions for Protoplast Isolation and Regeneration of Hyperproducing Cellulase Recombinants

作　　者：尹睿[1] 陈红漫[1] 张成刚[2] 

机构地区：[1]沈阳农业大学,辽宁沈阳110161 [2]中科院沈阳应用生态研究所,辽宁沈阳110015

出　　处：《沈阳农业大学学报》2000年第6期556-558,共3页Journal of Shenyang Agricultural University

摘　　要：研究了4种营养缺陷型产纤维素酶菌的原生质体形成和再生条件。结果表明 ,它们原生质体形成条件均以0.2mol·L-1(pH5.8)磷酸缓冲液为好。绿色木霉(Trichodermaviride)UV2 -11 和另一未定木霉(T.sp.)UV2 -2 以0.7mol·L-1KCl的渗透稳定剂为好 ,菌丝菌龄分别为20,18～20h ;康宁木霉(T.Koningii)UV2 -1 和UV2 -15 以0.6mol·L -1NaCl的渗透稳定剂为好 ,菌丝菌龄均为16～20h,酶解时间均为2～3h。原生质体再生培养基以加入0.5%酵母膏和0.5 %泛酸钙的改良Czapek培养基效果好 ,用0.7mol·L-1 KCl和NaCl作为渗透稳定剂 。The optimal conditions for protoplast isolation and regeneration of 4 auxotrophic mutant strains of Trichoderma were studied. 0.2mol·L-1 (pH5.8) Phosphate buffer is the better buffer system for protoplast isolation of all 4 strains. The best osmotic stablizer for T. Viride UV2-11 and T. sp UV2-2 is 0.7mol·L-1 KCl, and the best mycelia age is 20 and 18～20h, respectly. The best osmotic stablizer for T. Koningii UV2-1 and UV2-15 is 0.6mol·L-1 NaCl and the best mycelia age is 16～20 hours. The suitable enzymolysis time is 2～3h. Czapek medium containing 0.5% yeast extract and 0.5% Calium pantotthenic is the suitable regeneration medium for all 4 strains and both 0.7mol·L-1 KCl and 0.6mol·L-1 NaCl is the best osmotic stablizer for protoplast regeneration.

关 键 词：康宁木霉 绿色木霉 原生质体 再生条件 形成条件 纤维毒酶 缓冲液-渗透稳定剂系统 酶解时间 工业微生物 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]