IMMUNOAFFINITY PURIFICATION AND COMPARISON OF ALLANTOINASES FROM SOYBEAN ROOT-NODULES AND COTYLEDONS

被引:11
作者
BELL, JA [1 ]
WEBB, MA [1 ]
机构
[1] PURDUE UNIV, DEPT BOT & PLANT PATHOL, W LAFAYETTE, IN 47907 USA
关键词
D O I
10.1104/pp.107.2.435
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Allantoinase (allantoin amidohydrolase, EC 3.5.2.5) catalyzes the conversion of allantoin to allantoic acid in the final step of ureide biogenesis. We have purified allantoinase more than 4000-fold by immunoaffinity chromatography from root nodules and cotyledons of soybean (Glycine max [L.] Merr.). We characterized and compared properties of the enzyme from the two sources. Seed and nodule allantoinases had 80% identity in the first 24 amino acid residues of the N terminus. Two-dimensional gel electrophoresis of the purified enzymes showed that multiple forms were present in each. Allantoinases from nodules and cotyledons had very low affinity for allantoin with a K-m for allantoin of 17.3 mM in cotyledons and 24.4 mM in nodules. Both had activity in a broad range of pH values from 6.5 to 7.5. In addition, purified allantoinase from both sources was very heat stable. Enzyme activity was stable after 1 h at 70 degrees C, decreased gradually with heating to 85 degrees C, and was lost at 90 to 95 degrees C. Although these studies have revealed some differences between allantoinases in seeds and nodules, the differences were not reflected in key enzyme properties. The immunoaffinity approach enabled purification of allantoinase from soybean root nodules and simplified its purification from cotyledons, thereby allowing characterization and comparison of the enzyme from the two sources.
引用
收藏
页码:435 / 441
页数:7
相关论文
共 45 条
[1]  
AMARJIT SR, 1985, PHYTOCHEMISTRY, V24, P415
[2]   MECHANISMS OF THERMOPHILY [J].
AMELUNXEN, RE ;
MURDOCK, AL .
CRC CRITICAL REVIEWS IN MICROBIOLOGY, 1978, 6 (04) :343-393
[3]   EVIDENCE FOR A PURINE PATHWAY OF UREIDE SYNTHESIS IN N2-FIXING NODULES OF COWPEA [VIGNA-UNGUICULATA (L) WALP] [J].
ATKINS, CA ;
RAINBIRD, R ;
PATE, JS .
ZEITSCHRIFT FUR PFLANZENPHYSIOLOGIE, 1980, 97 (03) :249-260
[4]   PURINE BIOSYNTHESIS AND CATABOLISM IN SOYBEAN ROOT-NODULES - INCORPORATION OF C-14 FROM (CO2)-C-14 INTO XANTHINE [J].
BOLAND, MJ ;
SCHUBERT, KR .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1982, 213 (02) :486-491
[5]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]   THE ALLANTOINASE (DAL1) GENE OF SACCHAROMYCES-CEREVISIAE [J].
BUCKHOLZ, RG ;
COOPER, TG .
YEAST, 1991, 7 (09) :913-923
[7]   PURIFICATION AND PROPERTIES OF 2 FORMS OF GLUTAMINE-SYNTHETASE FROM THE PLANT FRACTION OF PHASEOLUS ROOT-NODULES [J].
CULLIMORE, JV ;
LARA, M ;
LEA, PJ ;
MIFLIN, BJ .
PLANTA, 1983, 157 (03) :245-253
[8]  
DAMSZ B, 1994, PLANT CELL PHYSIOL, V35, P979
[9]  
FRANKE W, 1965, ENZYMOLOGIA, V29, P251
[10]   POSSIBLE ROLE OF ALLANTOIN AND INFLUENCE OF NODULATION ON ITS PRODUCTION IN SOYBEAN PLANTS [J].
FUJIHARA, S ;
YAMAMOTO, K ;
YAMAGUCHI, M .
PLANT AND SOIL, 1977, 48 (01) :233-242