Identification of Missing Genes and Enzymes for Autotrophic Carbon Fixation in
            <i>Crenarchaeota</i>

Ramos‐Vera, W. Hugo; Weiss, Michael J.; Strittmatter, Eric F.; Kockelkorn, Daniel; Fuchs, Georg

doi:10.1128/jb.01156-10

Cited by 50 publications

(59 citation statements)

References 54 publications

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“…The genes that were incorporated into P. furiosus to enable it to use carbon dioxide are the first part of the 3-HP/4-hydroxybutyrate (4-HB) pathway of M. sedula, which consists of 13 enzymes in total (17). In one turn of the cycle, two molecules of carbon dioxide are added to one molecule of acetyl coenzyme A (acetyl-CoA) to generate a second molecule of acetyl-CoA (Fig.…”

Section: Resultsmentioning

confidence: 99%

Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide

Keller

Schut

Lipscomb

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

119

103

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Microorganisms can be engineered to produce useful products, including chemicals and fuels from sugars derived from renewable feedstocks, such as plant biomass. An alternative method is to use low potential reducing power from nonbiomass sources, such as hydrogen gas or electricity, to reduce carbon dioxide directly into products. This approach circumvents the overall low efficiency of photosynthesis and the production of sugar intermediates. Although significant advances have been made in manipulating microorganisms to produce useful products from organic substrates, engineering them to use carbon dioxide and hydrogen gas has not been reported. Herein, we describe a unique temperature-dependent approach that confers on a microorganism (the archaeon Pyrococcus furiosus, which grows optimally on carbohydrates at 100°C) the capacity to use carbon dioxide, a reaction that it does not accomplish naturally. This was achieved by the heterologous expression of five genes of the carbon fixation cycle of the archaeon Metallosphaera sedula, which grows autotrophically at 73°C. The engineered P. furiosus strain is able to use hydrogen gas and incorporate carbon dioxide into 3-hydroxypropionic acid, one of the top 12 industrial chemical building blocks. The reaction can be accomplished by cell-free extracts and by whole cells of the recombinant P. furiosus strain. Moreover, it is carried out some 30°C below the optimal growth temperature of the organism in conditions that support only minimal growth but maintain sufficient metabolic activity to sustain the production of 3-hydroxypropionate. The approach described here can be expanded to produce important organic chemicals, all through biological activation of carbon dioxide.

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Section: Resultsmentioning

confidence: 99%

Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide

Keller

Schut

Lipscomb

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

119

103

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“…9) Another ST0064 ortholog of Thermoproteus neutrophilus (Tneu 0421, 46 and 69% amino acid sequence identity and similarity) shows succinyl-CoA reductase activity. 10,11) An ortholog of Methanocaldococcus jannaschii (MJ 1411, 42 and 64% amino acid sequence identity and similarity) shows lactaldehyde dehydrogenase activity with broad substrate specificity. It can utilize propionaldehyde, glyceraldehyde (GA), and crotonaldehyde in addition to lactaldehyde, but no GAP oxidation activity was detected.…”

mentioning

confidence: 99%

Archaeal Aldehyde Dehydrogenase ST0064 fromSulfolobus tokodaii, a Paralog of Non-Phosphorylating Glyceraldehyde-3-phosphate Dehydrogenase, Is a Succinate Semialdehyde Dehydrogenase

Ito

Chishiki

Fushinobu

et al. 2013

Bioscience, Biotechnology, and Biochemistry

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Aldehyde dehydrogenase ST0064, the closest paralog of previously characterized allosteric non-phosphorylating glyceraldehyde-3-phosphate (GAP) dehydrogenase (GAPN, ST2477) from a thermoacidophilic archaeon, Sulfolobus tokodaii, was expressed heterologously and characterized in detail. ST0064 showed remarkable activity toward succinate semialdehyde (SSA) (K m of 0.0029 mM and k cat of 30.0 s À1 ) with no allosteric regulation. Activity toward GAP was lower (K m of 4.6 mM and k cat of 4.77 s À1 ), and previously predicted succinyl-CoA reductase activity was not detected, suggesting that the enzyme functions practically as succinate semialdehyde dehydrogenase (SSADH). Phylogenetic analysis indicated that archaeal SSADHs and GAPNs are closely related within the aldehyde dehydrogenase superfamily, suggesting that they are of the same origin.

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“…Autotrophy in M. sedula uses a recently discovered pathway for carbon fixation (6). Stable carbon isotope fractionation has shown that bicarbonate HCO 3 Ϫ is used as an inorganic carbon source through the 3-hydroxypropionate/4-hydroxypropionate pathway (6,32). M. sedula is also considerably more resistant to metals than chemoheterotrophic Sulfolobus species.…”

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confidence: 99%

Metal Resistance and Lithoautotrophy in the Extreme Thermoacidophile Metallosphaera sedula

et al. 2012

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Archaea such as Metallosphaera sedula are thermophilic lithoautotrophs that occupy unusually acidic and metal-rich environments. These traits are thought to underlie their industrial importance for bioleaching of base and precious metals. In this study, a genetic approach was taken to investigate the specific relationship between metal resistance and lithoautotrophy during biotransformation of the primary copper ore, chalcopyrite (CuFeS 2 ). In this study, a genetic system was developed for M. sedula to investigate parameters that limit bioleaching of chalcopyrite. The functional role of the M. sedula copRTA operon was demonstrated by cross-species complementation of a copper-sensitive Sulfolobus solfataricus copR mutant. Inactivation of the gene encoding the M. sedula copper efflux protein, copA, using targeted recombination compromised metal resistance and eliminated chalcopyrite bioleaching. In contrast, a spontaneous M. sedula mutant (CuR1) with elevated metal resistance transformed chalcopyrite at an accelerated rate without affecting chemoheterotrophic growth. Proteomic analysis of CuR1 identified pleiotropic changes, including altered abundance of transport proteins having AAA-ATPase motifs. Addition of the insoluble carbonate mineral witherite (BaCO 3 ) further stimulated chalcopyrite lithotrophy, indicating that carbon was a limiting factor. Since both mineral types were actively colonized, enhanced metal leaching may arise from the cooperative exchange of energy and carbon between surface-adhered populations. Genetic approaches provide a new means of improving the efficiency of metal bioleaching by enhancing the mechanistic understanding of thermophilic lithoautotrophy.N early 80% of current global copper reserves are comprised of low-quality metal ores (10,25,44). Consequently, the need for cost-efficient extraction methods has promoted interest in the use of microbe-based processing. Bioleaching is an established approach for the extraction of base and precious metals from sulfidic ores (30,34,35,43). Bioleaching using thermophilic microbes is of particular importance for certain metals. In the case of copper, elevated temperatures produced naturally in heaps overcome recalcitrant extraction due to surface passivation while chemical reaction rates are accelerated (29,30). A critical disadvantage of bioleaching is the amount of time required for metal solubilization, often spanning years (22,38). Therefore, improved metal recovery must include factors that accelerate this process, particularly those inspired by biotechnologic approaches (48).Thermoacidophilic archaea include taxa that are lithoautotrophic and unusually metal resistant (3,19). These organisms are native to pyritic or sulfur-rich geothermal habitats and are used to recover base and precious metals from low-quality sulfidic minerals through bioleaching processes (31). In the bioleaching process, both direct and indirect leaching mechanisms have been proposed that convert metals to soluble forms (33, 40). Metal release may occur via metab...

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Identification of Missing Genes and Enzymes for Autotrophic Carbon Fixation in Crenarchaeota

Cited by 50 publications

References 54 publications

Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide

Exploiting microbial hyperthermophilicity to produce an industrial chemical, using hydrogen and carbon dioxide

Archaeal Aldehyde Dehydrogenase ST0064 fromSulfolobus tokodaii, a Paralog of Non-Phosphorylating Glyceraldehyde-3-phosphate Dehydrogenase, Is a Succinate Semialdehyde Dehydrogenase

Metal Resistance and Lithoautotrophy in the Extreme Thermoacidophile Metallosphaera sedula

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