DCO Project Summary

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Project Title
Metabolic Reconstruction of Desulforudis audaxviator, a Subsurface Sentinel Species
Start DateEnd Date
2014-06-01 2014-12-31
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The subsurface sentinel species Candidatus Desulforudis audaxviator is a bacterium in the Firmicutes phylum and forms a unique one-species ecosystem in deep-fracture hot aquifers. So far, Desulforudis and its relatives have resisted persistent efforts at cultivation and thus their growth physiology and capacity for carbon transformation remain speculative. This project aims to initiate reconstruction of the C utilization pathway of this strain in a CO-utilizing surface hot spring model organism in order to obtain specific data on C uptake and processing systems. The key subsurface strains are known only by their genomes, which inform us that they are strict anaerobes and that the Wood-Ljungdahl Pathway generating Acetyl CoA from carbon monoxide is likely to be an important route for primary carbon acquisition. Our approach could yield information critical to isolation of Cand. D. audaxviator, leading to subsequent projects in collaboration with Drs. Onstott and Moser. The long-term goal of this consortium will be to isolate and characterize deep-dwelling strains of Firmicutes and other extremophiles.


Our objectives include:

1. Obtain insights into the temperature range of D. audaxviator by studying the thermoactivity of two recombinant enzymes expressed from genes deduced from its genome.

2. Complement the deficient C1 utilization pathway in the model organism with a carbon monoxide dehydrogenase from D. audaxviator.


We will achieve these objectives by synthesizing genes encoding the Carbon Monoxide Dehydrogenase and the Group III Chaperonin, cloning genes into shuttle vectors for expression in E. coli and in a model organism, the CO-utilizing thermophile Carboxydothermus hydrogenoformans and by purifying and characterizing recombinant Cpn60 and CODH enzymes. The work applies directly to Decadal Goal II, "Determine the principal pathways of carbon transformations in the subsurface and quantify the rates of these reactions." Further study of the metagenomes or metatranscriptomes from well effluents will have diminishing returns of relevant data on the rates and mechanisms of carbon biotransformation unless key species can be successfully isolated and their primary carbon fixation pathways studied.

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