This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Berman, M H Articles by Frazer, A C Search for Related Content PubMed PubMed Citation Articles by Berman, M H Articles by Frazer, A C Agricola Articles by Berman, M H Articles by Frazer, A C

 Previous Article  |  Next Article 

Appl Environ Microbiol. 1992 March; 58(3): 925-931

Importance of tetrahydrofolate and ATP in the anaerobic O-demethylation reaction for phenylmethylethers.

Department of Microbiology, New York University Medical Center, New York 10016.

ABSTRACT

DL-Tetrahydrofolate (THF) and ATP were necessary for the anaerobic O-demethylation of phenylmethylethers in cell extracts of the type strain (ATCC 29683) of the homoacetogen Acetobacterium woodii. The reactants for this enzymatic activity have not been previously demonstrated in any system, nor has the mediating enzyme been studied. An assay using reaction mixtures containing 1 mM THF, 2 mM ATP, and 2 mM hydroferulate (i.e., 4-hydroxy,3-methoxyphenylpropionate) was developed and was performed under stringent anaerobic conditions. Pyridine nucleotides and several other possible cofactors were tested but had no effect on the activity. After centrifugation of disrupted cells at 27,000 x g, the activity was found primarily in the supernatant, which had a specific activity of 14.2 +/- 0.5 nmol/min/mg of protein. At saturating levels of each of the other two substrates, apparent Km values for the variable substrate were 0.65 mM hydroferulate, 0.27 mM ATP, and 0.17 mM THF. Activity was significantly decreased when extract was preincubated at 60 degrees C and was completely lost after preincubation in air for 30 min. Thus, the soluble anaerobic O-demethylating enzyme system of A. woodii is oxygen sensitive. The THF- and ATP-dependent activity measurable in the soluble fraction of cell extracts constituted about 34% of the activity seen with intact cells.

This article has been cited by other articles:

• Abe, T., Masai, E., Miyauchi, K., Katayama, Y., Fukuda, M. (2005). A Tetrahydrofolate-Dependent O-Demethylase, LigM, Is Crucial for Catabolism of Vanillate and Syringate in Sphingomonas paucimobilis SYK-6. J. Bacteriol. 187: 2030-2037 [Abstract] [Full Text]  
• Masai, E., Sasaki, M., Minakawa, Y., Abe, T., Sonoki, T., Miyauchi, K., Katayama, Y., Fukuda, M. (2004). A Novel Tetrahydrofolate-Dependent O-Demethylase Gene Is Essential for Growth of Sphingomonas paucimobilis SYK-6 with Syringate. J. Bacteriol. 186: 2757-2765 [Abstract] [Full Text]  
• Naidu, D., Ragsdale, S. W. (2001). Characterization of a Three-Component Vanillate O-Demethylase from Moorella thermoacetica. J. Bacteriol. 183: 3276-3281 [Abstract] [Full Text]  
• Sonoki, T., Obi, T., Kubota, S., Higashi, M., Masai, E., Katayama, Y. (2000). Coexistence of Two Different O Demethylation Systems in Lignin Metabolism by Sphingomonas paucimobilis SYK-6: Cloning and Sequencing of the Lignin Biphenyl-Specific O-Demethylase (LigX) Gene. Appl. Environ. Microbiol. 66: 2125-2132 [Abstract] [Full Text]  
• Nishikawa, S., Sonoki, T., Kasahara, T., Obi, T., Kubota, S., Kawai, S., Morohoshi, N., Katayama, Y. (1998). Cloning and Sequencing of the Sphingomonas (Pseudomonas) paucimobilis Gene Essential for the O Demethylation of Vanillate and Syringate. Appl. Environ. Microbiol. 64: 836-842 [Abstract] [Full Text]