CHLOROCATECHOL 1,2-DIOXYGENASE FROM RHODOCOCCUS-ERYTHROPOLIS 1CP - KINETIC AND IMMUNOCHEMICAL COMPARISON WITH ANALOGOUS ENZYMES FROM GRAM-NEGATIVE STRAINS

被引:49
作者
MALTSEVA, OV [1 ]
SOLYANIKOVA, IP [1 ]
GOLOVLEVA, LA [1 ]
机构
[1] RUSSIAN ACAD SCI, INST BIOCHEM & PHYSIOL MICROORGANISMS, PUSHCHINO, RUSSIA
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1994年 / 226卷 / 03期
关键词
D O I
10.1111/j.1432-1033.1994.01053.x
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
Chlorocatechol 1,2-dioxygenase from Rhodococcus erq,thropolis 1CP was purified to homogeneity. In contrast to chlorocatechol 1,2-dioxygenase from Gram-negative strains which have a very broad substrate tolerance, the Rhodococcus enzyme was relatively more specific and had a distinct preference for 4-substituted catechols. Protein and metal analysis indicate an unusual stoichiometry of one atom each of iron and manganese/64-kDa homodimer. The N-terminal amino acid sequence (27 residues) of the Rhodococcus chlorocatechol 1,2-dioxygenase was determined and exhibited 15-22% identity to the published sequences of catechol 1,2-dioxygenases and other chlorocatechol 1,2-dioxygenases. Antiserum was raised in rabbits and antibodies against Rhodococcus chlorocatechol 1,2-dioxygenase were affinity purified. Dot-blot analysis revealed a very weak reaction between the antibodies and partially purified chlorocatechol 1,2-dioxygenases from Alcaligenes eutrophus JMP134 and Pseudomonas putida 87. No reaction between these antibodies and above enzymes was observed using Western blotting. Kinetic and immunochemical data as well as comparison of subunit molecular mass and suggest that the Rhodococcus enzyme differs significantly from the known highly similar chlorocatechol 1,2-dioxygenases of Gram-negative strains and seems to be only distantly related to them.
引用
收藏
页码:1053 / 1061
页数:9
相关论文
共 37 条