几种固定化苯丙氨酸解氨酶方法的比较

A Comparison of Phenylalanine Ammonia- lyases Immobilized by Different Methods

摘要: 目的　对不同方法固定化苯丙氨酸解氨酶 (PAL)的偶联率或包被率以及固定化对PAL活性和稳定性的影响进行比较, 以期得到较为方便、有效的固定化 PAL。方法　 (1 )聚丙烯酰胺凝胶 (PAG)包埋 PAL;(2 )载体偶联法将 PAL固定于 Bio- Gel P- 30 0 ;(3)将 PAL包被于三种脂类组成不同的质脂体 ;(4)对诱变选育出的含 PAL活性酵母细胞进行丙酮处理。结论　 (1 )固定化酶活性采用 T∶C比为 8∶ 30明显高于 8∶ 1 9;(2 )以戊二醛将 PAL固定于 Bio- Gel P30 0优于用DCD固定化的酶 ;(3)质脂体包被中以脱水再加水法对酶活性影响较少, 脂类组成以磷脂酰胆碱 :胆固醇 :磷脂酰丝氨酸比值 7∶ 2∶ 1最好 ;(4)固定化 PAL活性酵母细胞能较好地抵抗人工肠液中胰蛋白酶的水解作用, 亦能明显降低苯丙氨酸的浓度。

Abstract: Objective Phenglalanine Ammonia-lyases (PAL) from mung bean was immobilized by different. Methods Entrapped by PAG or liposome.Immobilized by covalent coupling on Bio-Gel P-300.Immobilized and permeated PAL active yeast cells were obtained by acetone treatment.The percentage of the PAL entrapped and enzyme activity retained were compared. Conclusion (1)PAL is better entrapped by PAG with a T∶C ratio of 8∶30.(2)PAL immobilized by glutaric dialdehyde on Bio-Gel P-300 is better than by CDC.(3)The best one i...