ISOLATION AND CHARACTERIZATION OF GLYCOLIC ACID DEHYDROGENASE FROM HUMAN-LIVER

被引:49
作者
FRY, DW [1 ]
RICHARDSON, KE [1 ]
机构
[1] OHIO STATE UNIV,DEPT PHYSIOL CHEM,COLUMBUS,OH 43210
关键词
(Human liver); Glycolic acid dehydrogenase;
D O I
10.1016/0005-2744(79)90134-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Glycolic acid dehydrogenase has been purified over 800-fold from human liver by (NH4)2SO4 fractionation and column chromatography with DEAE-cellulose and hydroxyapatite. The enzyme catalyzes the direct oxidation of glycolate to oxalate without forming glyoxylate as a free intermediate. Activity is found only in the liver in the soluble fraction. The enzyme is specific for glycolate and inhibits no activity towards glycine or glyoxylate. Glyoxylate and dl-phenyllactate exhibit the enzyme. Optimum activity occurs sharply at pH 6.1 and the Michaelis constant for glycolate was 6.3 · 10-5M. Molecular oxygen does not appear to be the electron acceptor and no requirement for cofactors has been demonstrated, although flavin mononucleotide, ascorbate and cytochrome c stimulate activity. The isolation of this enzyme which may account for a significant part of the normal oxalate excretion in man, provides a more complete understanding of the pathways of oxalate biosynthesis and must be taken into account when considering possible methods for controlling disorders of oxalate metabolism. © 1979.
引用
收藏
页码:482 / 491
页数:10
相关论文
共 25 条
[3]   TISSUE FRACTIONATION STUDIES .6. INTRACELLULAR DISTRIBUTION PATTERNS OF ENZYMES IN RAT-LIVER TISSUE [J].
DUVE, CD ;
PRESSMAN, BC ;
GIANETTO, R ;
WATTIAUX, R ;
APPELMANS, F .
BIOCHEMICAL JOURNAL, 1955, 60 (1-4) :604-617
[4]   ENZYMATIC OXALATE DECARBOXYLATION IN ASPERGILLUS NIGER [J].
EMILIANI, E ;
BEKES, P .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1964, 105 (03) :488-&
[5]   PATHWAYS OF OXALATE FORMATION FROM PHENYLALANINE, TYROSINE, TRYPTOPHAN AND ASCORBIC-ACID IN RAT [J].
GAMBARDELLA, RL ;
RICHARDSON, KE .
BIOCHIMICA ET BIOPHYSICA ACTA, 1977, 499 (01) :156-168
[6]  
GIBBS DA, 1966, CLIN SCI, V31, P285
[7]  
HAGLER L, 1973, AM J CLIN NUTR, V26, P758
[8]  
Jorns M S, 1975, Methods Enzymol, V41, P337
[9]  
KUN E, 1954, J BIOL CHEM, V210, P269
[10]   SYNTHESIS OF OXALATE FROM HYDROXYPYRUVATE BY ISOLATED PERFUSED RAT-LIVER - MECHANISM OF HYPEROXALURIA IN L-GLYCERIC ACIDURIA [J].
LIAO, LL ;
RICHARDSON, KE .
BIOCHIMICA ET BIOPHYSICA ACTA, 1978, 538 (01) :76-86