|本期目录/Table of Contents|

[1]李兴德,路英华.转化血型用蕃茄α-D-半乳糖苷酶的分离、纯化及理化性质[J].应用与环境生物学报,1996,2(04):369-375.
 Li Xingde,Lu Yinghua.PURIFICATION AND CHARACTERIZATION OF α-D-GALACTOSIDASE FROM TOMATO[J].Chinese Journal of Applied & Environmental Biology,1996,2(04):369-375.
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转化血型用蕃茄α-D-半乳糖苷酶的分离、纯化及理化性质()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
2卷
期数:
1996年04期
页码:
369-375
栏目:
论文
出版日期:
1996-11-25

文章信息/Info

Title:
PURIFICATION AND CHARACTERIZATION OF α-D-GALACTOSIDASE FROM TOMATO
作者:
李兴德1 路英华2
1. 中国学科院成都生物研究所, 成都 610041;
2. 华东师范大学生物系, 上海 200062
Author(s):
Li Xingde Lu Yinghua
1. Chengdu Institute of Biology, Academia Sinica, Chengdu 610041;
2. Department of Biology, East China Normal University, Shanghai 200062
关键词:
血型转化蕃茄α-D-半乳糖苷酶分离纯化理化性质
Keywords:
blood group conversiontomatoα-D-galactosidasepurilictionphysicochemical characters
摘要:
成熟蕃茄匀浆后,经硫酸铵盐析,DEAE-SephadexA-50离子交换层析,SepadexG-100凝胶过滤和Melibiose-Agarose亲和层析,获得了α-D-半乳糖苷酶(C.E.3.2.1.11).酶制剂经PAGE检测为一条带;SDS-G-PAGE测得酶Mr为34000;比活力52.9U/mg·;提纯倍数为52901产率为45%.酶专-催化以α-D-半乳糖为末端a-(1,3)连接的糖苷键,以PNPG(对硝基苯-α-D-半乳糖昔)为废物,酶催化反应的Km=0.11mmol/L,Vmax为67μmol·mg1-·min-1.t稳定范是0~35℃;PH稳定范围是4.0~7.0.最适pH为5.1.半乳糖是酶的竞争性抑制剂;Cu2+、Zn2+、Mn2+、Fe3+、Ag+和EDTA对酶活性无影响。纯酶制剂可作为B型血向O型血转化的工具酶液。
Abstract:
Tomato(Lycoporiscoum) was selected as raw material for purilyins α-D-galactosidase.Purified tomatoα-D-galactosidase was obtained by (NH4)2SO4 treatment,DEAE-Sephadex A-50 ion-exchange chromatography,Sephadex G-100 gel filtration and Melibiose-agnrose affinity chromatography.The results showed thata 5 290 fold purification of enzyme was obtained, its recovery rate was 45%,and the specific activity was52.9 U/mg.pro.The enzyme’s M, was 34 000 by SDS-G-PAGE.The tomato α-D-galactosidase was stableat pH 4.0-7.0 and tempetature 0-35℃.The enzyme hydrolysed substrate PNPG with the Km 0.11mmol/L,V,max = 67μmol·mg-1· min-1,optimum pH 5.1.The α-D-galactose is a competitive inhibitor ofthe enzyme.Metal ions CU2+.Zn2+.Mn2+.Fe3+.Ag+ and EDTA have no effects upon enzymology activity.

参考文献/References:

[1] 上海生物制品研究所血型组编.血型与血库.上海:上海人民出版社,1977
[2] Yatiziv S.Action of a-D-galactosidase on glyeopeotein from humn B-erythrocytes.Biochem Riophys R Conuntrn.1971,45(2):123-126
[3] Harpaz N,Flowers H M,Sharon N.Studies on B-antigenic sites of humn erthtocytes by use of coffee bean α-D-galactosidase.Arch Biochem Biophys 1975,170:676-683
[4] Dybus S.Action of α-D-galactosidase from clostridiumsporogenes and coffee bean on blood peoup B antigen of erythricytes.Tans,fusion.1983,23(3):78-86
[5] Barham D.Studies on the distribution of α-D-galactosidase in seeds.Photochemisty.1971,10:1759-1763
[6] Harpaz N,Flowers H M,Sharon N.Purification of coffee bean α-D-galactosidase by affinity chromatography.Biochem Biophys Acla.1974,341:213-216
[7] Kandiiah Balasubtamanian.Purification of α-D-galactosidase from coconut.Photochemistry.1986,25(8):1819-1821
[8] iton Tomoko,Yutaka Uda,Hiroki Nakagwa.Purification and characterization of α-D-galactosidase from watermelon.J Biochem (Tokyo).1986,99(1):243-250
[9] Jack Goldstein.Group B erythrocytes enzymatically converted to group O surive normaVy in A,B and O individuals.Sciersce.1982,215:103-105
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备注/Memo

备注/Memo:
收稿日期:1996-2-9;接受日期:1996-4-30。
基金项目:国家自然科学基金;中国科学院成都地奥科学基金(DASF)资助项目
更新日期/Last Update: 1900-01-01