Affiliation:
1. Institut Biologie II, Mikrobiologie, Albert-Ludwigs-Universität Freiburg, Freiburg, Germany
Abstract
ABSTRACT
The anaerobic metabolism of phenol in the beta-proteobacterium
Thauera aromatica
proceeds via
para
-carboxylation of phenol (biological Kolbe-Schmitt carboxylation). In the first step, phenol is converted to phenylphosphate which is then carboxylated to 4-hydroxybenzoate in the second step. Phenylphosphate formation is catalyzed by the novel enzyme phenylphosphate synthase, which was studied. Phenylphosphate synthase consists of three proteins whose genes are located adjacent to each other on the phenol operon and were overproduced in
Escherichia coli
. The promoter region and operon structure of the phenol gene cluster were investigated. Protein 1 (70 kDa) resembles the central part of classical phosphoenolpyruvate synthase which contains a conserved histidine residue. It catalyzes the exchange of free [
14
C]phenol and the phenol moiety of phenylphosphate but not the phosphorylation of phenol. Phosphorylation of phenol requires protein 1, MgATP, and another protein, protein 2 (40 kDa), which resembles the N-terminal part of phosphoenol pyruvate synthase. Proteins 1 and 2 catalyze the following reaction: phenol + MgATP + H
2
O→phenylphosphate + MgAMP + orthophosphate. The phosphoryl group in phenylphosphate is derived from the β-phosphate group of ATP. The free energy of ATP hydrolysis obviously favors the trapping of phenol (
K
m
, 0.04 mM), even at a low ambient substrate concentration. The reaction is stimulated severalfold by another protein, protein 3 (24 kDa), which contains two cystathionine-β-synthase domains of unknown function but does not show significant overall similarity to known proteins. The molecular and catalytic features of phenylphosphate synthase resemble those of phosphoenolpyruvate synthase, albeit with interesting modifications.
Publisher
American Society for Microbiology
Subject
Molecular Biology,Microbiology
Reference51 articles.
1. Aiba, H., S. Adhya, and B. de Combrugge. 1981. Evidence for two functional gal promoters in intact Escherichia coli. J. Biol. Chem.256:11905-11910.
2. Anders, H. J., A. Kaetzke, P. Kämpfer, W. Ludwig, and G. Fuchs. 1995. Taxonomic position of aromatic-degrading denitrifying pseudomonad strains K 172 and KB 740 and their description as new members of the genera Thauera, as Thauera aromatica sp. nov., and Azoarcus, as Azoarcus evansii sp. nov., respectively, members of the beta subclass of the Proteobacteria. Int. J. Syst. Bacteriol.45:327-333.
3. Aresta, M., and A. Dibenedetto. 2002. Development of environmentally friendly syntheses: use of enzymes and biomimetic systems for the direct carboxylation of organic substrates. J. Biotechnol.90:113-128.
4. Baek, Y. H., and T. Nowak. 1982. Kinetic evidence for a dual role for muscle pyruvate kinase. Arch. Biochem. Biophys.217:491-497.
5. Bak, F., and F. Widdel. 1986. Anaerobic degradation of phenol and phenol derivatives by Desulfobacterium phenolicum sp. nov. Arch. Microbiol.146:177-180.
Cited by
65 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献