分别考察了液氮冷冻法、超声破碎法、甘氨酸与丙氨酸化学渗透法,不同化学渗透剂TritonX-100、Tween80、Span20、溴代十六烷基吡啶,有机溶剂甲苯、脲及EDTA对粘红酵母(Rhodotorula glutinis)胞内苯丙氨酸解氨酶(即PAL酶)释放及活力的影响,比较发现超声破碎与TritonX-100联用酶释放效果显著,先加入10%Triton X-100渗透6h,再在800W超声破碎30min,蛋白释放率达76.7%。总酶活70.3%,为提取胞内PAL酶提供了一种新方法。
The effects of freezing treatment by liquid nitrogen, ultrasonic disruption, chemical penetration by L-glycine and L-alanine, and some surfactants (TritonX-100, Tween80, Span20, Cetyl pyridine bromide, toluene, urea and EDTA) on the release of phenylalanine ammonia-lyase (PAL) in the cell of Rhodotorula glutinis were investigated in this article. The experiments indicated that the combination of ultrasonic disruption and TritonX-100 could release the PAL significantly. By penetrating with TritonX-100 for 6 h, then ultrasonic disruption under 800 W for 30 min, the release rate of protein reached 76.7%, the enzyme activity of PAL stabilized at 70.3% of the original total activity. The research provides a new method for the extraction of PAL in yeast ceils.