Novel formaldehyde-activating enzyme in Methylobacterium extorquens AM1 required for growth on methanol.
Formaldehyde is toxic for all organisms from bacteria to humans due to its reactivity with biological macromolecules. Organisms that grow aerobically on single-carbon compounds such as methanol and methane face a special challenge in this regard because formaldehyde is a central metabolic intermediate during methylotrophic growth. In the alpha-proteobacterium Methylobacterium extorquens AM1, we found a previously unknown enzyme that efficiently catalyzes the removal of formaldehyde: it catalyzes the condensation of formaldehyde and tetrahydromethanopterin to methylene tetrahydromethanopterin, a reaction which also proceeds spontaneously, but at a lower rate than that of the enzyme-catalyzed reaction. Formaldehyde-activating enzyme (Fae) was purified from M. extorquens AM1 and found to be one of the major proteins in the cytoplasm. The encoding gene is located within a cluster of genes for enzymes involved in the further oxidation of methylene tetrahydromethanopterin to CO(2). Mutants of M. extorquens AM1 defective in Fae were able to grow on succinate but not on methanol and were much more sensitive toward methanol and formaldehyde. Uncharacterized orthologs to this enzyme are predicted to be encoded by uncharacterized genes from archaea, indicating that this type of enzyme occurs outside the methylotrophic bacteria.
Vorholt
J A
JA
Max Planck Institute for Terrestrial Microbiology, 35043 Marburg, Germany. vorholt@mailer.uni-marburg.de
Marx
C J
CJ
Lidstrom
M E
ME
Thauer
R K
RK
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D019731:Carbon-Nitrogen Ligases / Q000145:classification / Q000378:metabolism*; D002384:Catalysis; D002874:Chromosome Mapping; D003470:Culture Media; D004789:Enzyme Activation / Q000187:drug effects; D005557:Formaldehyde / Q000378:metabolism* / Q000494:pharmacology; D019766:Genes, Archaeal; D000432:Methanol / Q000378:metabolism* / Q000494:pharmacology; D020601:Methylobacterium extorquens / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016296:Mutagenesis; D010641:Phenotype; D011622:Pterins / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013763:Tetrahydrofolates / Q000378:metabolism
D001426:Bacterial Proteins; D003470:Culture Media; D011622:Pterins; D013763:Tetrahydrofolates; D005557:Formaldehyde; C030371:5,6,7,8-tetrahydrofolic acid; C042416:5,6,7,8-tetrahydromethanopterin; D019731:Carbon-Nitrogen Ligases; C417713:Fae protein, Methylobacterium extorquens; D000432:Methanol