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Microbiology 146 (2000), 233-238
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Microbiology (2000), 146, 233-238.
© 2000 Society for General Microbiology

Physiology and Growth

Analysis of two formaldehyde oxidation pathways in Methylobacillus flagellatus KT, a ribulose monophosphate cycle methylotroph

Ludmila Chistoserdova1, Larissa Gomelskya,4, Julia A. Vorholt1,3, Mark Gomelskya,4, Yuri D. Tsygankov4 and Mary E. Lidstrom1,2

Departments of Chemical Engineering1 and Microbiology2, University of Washington, Seattle, WA 98195, USA
Max-Plank-Institut für Terrestrische Mikrobiologie, 35043 Marburg, Germany3
Institute of Genetics and Selection of Industrial Micro-organisms, 1st Dorozhny proezd 1, Moscow 113545, Russia4

Author for correspondence: Mary E. Lidstrom. Tel: +1 206 616 5282. Fax: +1 206 616 5721. e-mail: lidstrom{at}u.washington.edu

The roles of cyclic formaldehyde oxidation via 6-phosphogluconate dehydrogenase and linear oxidation via the tetrahydromethanopterin (H4MPT)-linked pathway were assessed in an obligate methylotroph, Methylobacillus flagellatus KT, by cloning, sequencing and mutating two chromosomal regions containing genes encoding enzymes specifically involved in these pathways:6-phosphogluconate dehydrogenase, glucose-6-phosphate dehydrogenase and methenyl H4MPT cyclohydrolase (gndA, zwf and mch). No null mutants were obtained in gndA or zwf, implying that the cyclic oxidation of formaldehyde is required for C1 metabolism in this obligate methylotroph, probably as the main energy-generating pathway. In contrast, null mutants were generated in mch, indicating that the H4MPT-linked pathway is dispensable. These mutants showed enhanced sensitivity to formaldehyde, suggesting that this pathway plays a secondary physiological role in this methylotroph. This function is in contrast to Methylobacterium extorquens AM1, in which the H4MPT-linked pathway is essential.

Keywords: methylotroph, ribulose monophosphate cycle, formaldehyde oxidation, tetrahydromethanopterin

Abbreviations: CH, cyclohydrolase; H4MPT, tetrahydromethanopterin; RMP, ribulose monophosphate

The GenBank accession numbers for the sequences of 2502 nt containing gndA and part of zwf, and of 2685 nt containing mch, are AF167580 and AF139592, respectively.

a Present address: Department of Molecular Biology, University of Wyoming, Laramie, WY 82071, USA.




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