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  • Incomplete Wood-Ljungdahl pathway facilitates one-carbon metabolism in organohalide-respiring Dehalococcoides mccartyi.

Incomplete Wood-Ljungdahl pathway facilitates one-carbon metabolism in organohalide-respiring Dehalococcoides mccartyi.

Proceedings of the National Academy of Sciences of the United States of America (2014-04-16)
Wei-Qin Zhuang, Shan Yi, Markus Bill, Vanessa L Brisson, Xueyang Feng, Yujie Men, Mark E Conrad, Yinjie J Tang, Lisa Alvarez-Cohen
ABSTRACT

The acetyl-CoA "Wood-Ljungdahl" pathway couples the folate-mediated one-carbon (C1) metabolism to either CO2 reduction or acetate oxidation via acetyl-CoA. This pathway is distributed in diverse anaerobes and is used for both energy conservation and assimilation of C1 compounds. Genome annotations for all sequenced strains of Dehalococcoides mccartyi, an important bacterium involved in the bioremediation of chlorinated solvents, reveal homologous genes encoding an incomplete Wood-Ljungdahl pathway. Because this pathway lacks key enzymes for both C1 metabolism and CO2 reduction, its cellular functions remain elusive. Here we used D. mccartyi strain 195 as a model organism to investigate the metabolic function of this pathway and its impacts on the growth of strain 195. Surprisingly, this pathway cleaves acetyl-CoA to donate a methyl group for production of methyl-tetrahydrofolate (CH3-THF) for methionine biosynthesis, representing an unconventional strategy for generating CH3-THF in organisms without methylene-tetrahydrofolate reductase. Carbon monoxide (CO) was found to accumulate as an obligate by-product from the acetyl-CoA cleavage because of the lack of a CO dehydrogenase in strain 195. CO accumulation inhibits the sustainable growth and dechlorination of strain 195 maintained in pure cultures, but can be prevented by CO-metabolizing anaerobes that coexist with D. mccartyi, resulting in an unusual syntrophic association. We also found that this pathway incorporates exogenous formate to support serine biosynthesis. This study of the incomplete Wood-Ljungdahl pathway in D. mccartyi indicates a unique bacterial C1 metabolism that is critical for D. mccartyi growth and interactions in dechlorinating communities and may play a role in other anaerobic communities.

MATERIALS
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Sigma-Aldrich
S-Acetyl-coenzyme A synthetase from baker's yeast (S. cerevisiae), lyophilized powder, ≥3 units/mg protein
Sigma-Aldrich
L-Serine, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
L-Serine, ReagentPlus®, ≥99% (HPLC)
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Carbon - Vitreous, rod, 100mm, diameter 3.0mm, glassy carbon
Carbon - Vitreous, rod, 5mm, diameter 3.0mm, glassy carbon
Carbon - Vitreous, tube, 100mm, outside diameter 10mm, inside diameter 3mm, wall thickness 3.5mm, glassy carbon
Carbon - Vitreous, foil, 100x100mm, thickness 1.0mm, glassy carbon
Carbon - Vitreous, foam, 300x300mm, thickness 30mm, bulk density 0.05g/cm3, porosity 96.5%
Carbon - Vitreous, foil, 50x50mm, thickness 4.0mm, glassy carbon
Carbon - Vitreous, foil, 100x100mm, thickness 6.0mm, glassy carbon
Carbon, foil, 100x100mm, thickness 0.2mm, flexible graphite, 99.8%
Carbon - Vitreous, foil, 50x50mm, thickness 1.0mm, glassy carbon
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Carbon - Vitreous, foil, 10mm disks, thickness 0.2mm, glassy carbon
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Carbon - Vitreous, rod, 200mm, diameter 3.0mm, glassy carbon
Carbon - Vitreous, rod, 200mm, diameter 7.0mm, glassy carbon
Carbon - Vitreous, foam, 300x300mm, thickness 20mm, bulk density 0.05g/cm3, porosity 96.5%
Carbon, foil, 100x100mm, thickness 1.0mm, flexible graphite, 99.8%
Carbon, foil, 100x100mm, thickness 0.125mm, rigid graphite, fine grain size, 99.95%
Carbon - Vitreous, foam, 150x150mm, 0.05g.cmué, porosity 96.5%, 24 pores/cm
Carbon, foil, 100x100mm, thickness 0.25mm, rigid graphite, fine grain size, 99.95%
Carbon, foil, 100x100mm, thickness 0.35mm, flexible graphite, 99.8%
Carbon - Vitreous, foil, 10x10mm, thickness 4.0mm, glassy carbon
Carbon, foil, 100x100mm, thickness 2.5mm, rigid graphite, fine grain size, 99.95%
Carbon - Vitreous, foam, 150x150mm, thickness 3.2mm, bulk density 0.05g/cm3, porosity 96.5%
Carbon - Vitreous, rod, 100mm, diameter 1.0mm, glassy carbon
Carbon, foil, 100x100mm, thickness 1.0mm, rigid graphite, fine grain size, 99.95%