绿豆苯丙氨酸解氨酶的分离提纯及抗肿瘤的初步研究  

The Isolation and Purification of Phenylalanine Ammonialyase from Green Grams

作　　者：牛三勇[1] 杜欣[2] 姚侃 

机构地区：[1]广东省老年医学研究所 [2]广东省人民医院

出　　处：《兰州医学院学报》1992年第3期148-151,共4页Journal of Lanzhou Medical College

摘　　要：采用丙酮沉淀,硫酸铵分段沉淀,超滤,DEAE-一纤维素柱层析。L-苯丙氨酸—琼脂糖4B亲和层析等方法,从绿豆芽中分离纯化苯丙氨酸解氨酶(PAL),纯化326倍,产率为9.6％。纯化的PAL经SDS聚丙烯酰胺凝胶电泳检验为单一的蛋白质区带。亚基分子量为72000道尔顿。绿豆的PAL有一定程度的耐热性。最适pH8.7,对酸碱波动耐受性强。经DEAE-纤维素柱层析从绿豆中分出二种PAL,其Km值分别为9.4×10^(-5)(PALl),6.2×10^(-5)(PAL2) 绿豆PAL对L1210小鼠淋巴细胞白血病细胞株的体外抑制实验,初步确定PAL对此瘤株的生长具有抑制作用。L-Phenylalanine ammonialyase (EC 4.3. 1.5. ; PAL) was isolated and purified from sprouting green grams (Phaseolus radiatus L.). The enzyme was purified from acetone powders prepared from sprout tissue raised for6 days by ammonium sulfate fractibnation, ultrafiltration, column chromatography on DEAL-cellulose and affinity chromatography on L-Phe-sepharose 4B. The enzyme was purified ca. 326-fold to essentially electrophoretic homogeneity with a yield of 9.6%. SDS-P AGE revealed a single stained protein band which corresponds to a subunit weight of 72000. The optimum P^H of the PAL isozyme were isolated from green gram sprout by colume chromatography on DEAE-cellulose namely PAL1 and PAL2. The Km value of PAL 19.4×10^(-5) M and 6.2×10^(-5) M respectively. The inhibition of the growth of murine leukemic lymphoblasts L1210 by PAL in vitro was noted. These results confirm C. W. Aell's studies and further support the concept that PAL exerts its inhibitory effect by depriving rapidly dividing cells of the essential amino acid phenylalanine.

关 键 词：解氨酶 绿豆 白血病细胞株 

分 类 号：R285.5[医药卫生—中药学]