Energy-conserving enzyme systems active during syntrophic acetate oxidation in the thermophilic bacterium Thermacetogenium phaeum
2019-11-29, Keller, Anja, Schink, Bernhard, Müller, Nicolai
The thermophilic acetogen Thermacetogenium phaeum uses the Wood-Ljungdahl pathway in both directions, either for the production of acetate from various compounds or for the oxidation of acetate in syntrophic cooperation with methanogens. In this study, energy conserving enzyme systems in Thermacetogenium phaeum were investigated in both metabolic directions. A gene cluster containing a membrane-bound periplasmically oriented formate dehydrogenase directly adjacent to putative menaquinone synthesis genes was identified in the genome. The protein products of these genes were identified by total proteome analysis, and menaquinone MK-7 had been found earlier as the dominant quinone in the membrane. Enzyme assays with membrane preparations and anthraquinone-2,6-disulfonate as electron acceptor verified the presence of a quinone-dependent formate dehydrogenase. A quinone-dependent methylene THF reductase is active in the soluble fraction and in the membrane fraction. From these results we conclude a reversed electron transport system from methyl-THF oxidation to CO2 reduction yielding formate as reduced product which is transferred to the methanogenic partner. The redox potential difference between methyl-THF (Eo’= -200 mV) and formate (Eo’= 432 mV) does not allow electron transfer through syntrophic formate removal alone. We postulate that part of the ATP conserved by substrate-level phosphorylation has to be invested into the generation of a transmembrane proton gradient by ATPase. This proton gradient could drive the endergonic oxidation of methyl-THF in an enzyme reaction similar to the membrane-bound reversed electron transport system previously observed in the syntrophically butyrate-oxidizing bacterium Syntrophomonas wolfei. To balance the overall ATP budget in acetate oxidation, we postulate that acetate is activated through an ATP-independent path via aldehyde:ferredoxin oxidoreductase (AOR) and subsequent oxidation of acetaldehyde to acetyl-CoA.
The use of Greek and Latin prepositions and prefixes in compound names: proposed emendation of Appendix 9 of the International Code of Nomenclature of Prokaryotes
2019-06, Oren, Aharon, Chuvochina, Maria, Schink, Bernhard
Part A of Appendix 9 - Orthography of the International Code of Nomenclature of Prokaryotes regulates the formation of compound generic names and specific epithets derived by combining two or more words or word elements of Latin and/or Greek origin, using the word stems and connecting vowels (-o- or -i-) following word elements derived from Greek and Latin, respectively. The rules given and the exceptions listed are suitable for substantives (nouns) and adjectives used as word elements, but not for prepositions and prefixes. Therefore, we propose a non-retroactive modification of Appendix 9 so that the guidelines given in Part A apply only to compound names that include a noun or an adjective in a non-final position. We also propose guidelines for the proper use of Greek and Latin prepositions, prefixes and adverbs in compound names in which the following word element starts with a vowel.
Pyrite formation from FeS and H2S is mediated through microbial redox activity
2019-04-02, Thiel, Joana, Byrne, James M., Kappler, Andreas, Schink, Bernhard, Pester, Michael
Pyrite is the most abundant iron−sulfur mineral in sediments. Over geological times, its burial controlled oxygen levels in the atmosphere and sulfate concentrations in seawater. However, the mechanism of pyrite formation in sediments is still being debated. We show that lithotrophic microorganisms can mediate the transformation of FeS and H2S to FeS2 at ambient temperature if metabolically coupled to methane-producing archaea. Our results provide insights into a metabolic relationship that could sustain part of the deep biosphere and lend support to the iron−sulfur-world theory that postulated FeS transformation to FeS2 as a key energy-delivering reaction for life to emerge.
Serious mismatches continue between science and policy in forest bioenergy
2019-11, Norton, Michael, Baldi, Andras, Buda, Vicas, Carli, Bruno, Cudlin, Pavel, Jones, Mike B., Korhola, Atte, Michalski, Rajmund, Novo, Francisco, Oszlányi, Július, Santos, Filpe Duarte, Schink, Bernhard, Shepherd, John, Vet, Louise, Walloe, Lars, Wijkman, Anders
In recent years, the production of pellets derived from forestry biomass to replace coal for electricity generation has been increasing, with over 10 million tonnes traded internationally—primarily between United States and Europe but with an increasing trend to Asia. Critical to this trade is the classification of woody biomass as ‘renewable energy’ and thus eligible for public subsidies. However, much scientific study on the net effect of this trend suggests that it is having the opposite effect to that expected of renewable energy, by increasing atmospheric levels of carbon dioxide for substantial periods of time. This review, based on recent work by Europe's Academies of Science, finds that current policies are failing to recognize that removing forest carbon stocks for bioenergy leads to an initial increase in emissions. Moreover, the periods during which atmospheric CO2 levels are raised before forest regrowth can reabsorb the excess emissions are incompatible with the urgency of reducing emissions to comply with the objectives enshrined in the Paris Agreement. We consider how current policy might be reformed to reduce negative impacts on climate and argue for a more realistic science‐based assessment of the potential of forest bioenergy in substituting for fossil fuels. The length of time atmospheric concentrations of CO2 increase is highly dependent on the feedstocks and we argue for regulations to explicitly require these to be sources with short payback periods. Furthermore, we describe the current United Nations Framework Convention on Climate Change accounting rules which allow imported biomass to be treated as zero emissions at the point of combustion and urge their revision to remove the risk of these providing incentives to import biomass with negative climate impacts. Reforms such as these would allow the industry to evolve to methods and scales which are more compatible with the basic purpose for which it was designed.
Naming classes of prokaryotes based on the rules of Latin grammar
2019-05, Oren, Aharon, Chuvochina, Maria, Schink, Bernhard, Ventura, Stefano
Recently a proposal was published to unify Rules 7, 8 and 9 of the International Code of Nomenclature of Prokaryotes. Based on this proposal, all names of taxa above the rank of genus must be in the feminine gender, the plural number. For the rank of class, this proposal contravenes Principle 3 of the Code, which states that the scientific names of all taxa are treated as Latin. The -ia ending of most names of classes belongs to nominative plural nouns of the neuter gender.
Alternative Pathways of Acetogenic Ethanol and Methanol Degradation in the Thermophilic Anaerobe Thermacetogenium phaeum
2019-03-19, Keller, Anja, Schink, Bernhard, Müller, Nicolai
Growth of the anaerobic thermophile Thermacetogenium phaeum with methanol, ethanol, ethanolamine, and acetate was investigated in axenic cultures and in syntrophic cultures with Methanothermobacter thermautotrophicus. Microcompartment genes were identified in the T. phaeum genome, and presence of microcompartments was confirmed by transmission electron microscopy and proteome analysis. These genes were expressed only during growth with ethanolamine. Proteome data were compared after growth with all four substrates, and activities of key enzymes of the Wood–Ljungdahl pathway and of enzyme systems leading to production or degradation of acetaldehyde such as alcohol dehydrogenase, aldehyde:ferredoxin oxidoreductase, acetate kinase, and phosphate acetyltransferase were measured in cytoplasmic fractions. Accounting of fermentation stoichiometries and growth yields with all four substrates showed that ethanol and methanol oxidation follow the same stoichiometries as in Acetobacterium woodii. On the other hand, the pathways of ethanol and methanol degradations vary between both organisms. Growth yields of T. phaeum were substantially lower than reported for A. woodii. Since T. phaeum has no Rnf complex encoded in its genome, the mechanisms of ATP synthesis have to be different from those of A. woodii. In addition to the central degradation pathways also found in A. woodii, T. phaeum maintains enzyme systems that compensate for the absence of an Rnf-complex but which on the other hand cause a loss of energy. On the basis of our data, pathways of methanol and ethanol degradation in T. phaeum are discussed.
Enzymes involved in phthalate degradation in sulphate-reducing bacteria
2019-10, Geiger, Robin Alexander, Junghare, Madan, Mergelsberg, Mario, Ebenau-Jehle, Christa, Jesenofsky, Vivien Jill, Jehmlich, Nico, von Bergen, Martin, Schink, Bernhard, Boll, Matthias
The complete degradation of the xenobiotic and environmentally harmful phthalate esters is initiated by hydrolysis to alcohols and o-phthalate (phthalate) by esterases. While further catabolism of phthalate has been studied in aerobic and denitrifying microorganisms, the degradation in obligately anaerobic bacteria has remained obscure. Here, we demonstrate a previously overseen growth of the δ-proteobacterium Desulfosarcina cetonica with phthalate/sulphate as only carbon and energy sources. Differential proteome and CoA ester pool analyses together with in vitro enzyme assays identified the genes, enzymes and metabolites involved in phthalate uptake and degradation in D. cetonica. Phthalate is initially activated to the short-lived phthaloyl-CoA by an ATP-dependent phthalate CoA ligase (PCL) followed by decarboxylation to the central intermediate benzoyl-CoA by an UbiD-like phthaloyl-CoA decarboxylase (PCD) containing a prenylated flavin cofactor. Genome/metagenome analyses predicted phthalate degradation capacity also in the sulphate-reducing Desulfobacula toluolica, strain NaphS2, and other δ-proteobacteria. Our results suggest that phthalate degradation proceeds in all anaerobic bacteria via the labile phthaloyl-CoA that is captured and decarboxylated by highly abundant PCDs. In contrast, two alternative strategies have been established for the formation of phthaloyl-CoA, the possibly most unstable CoA ester in biology.
Anaerobic degradation of xenobiotic isophthalate by the fermenting bacterium Syntrophorhabdus aromaticivorans
2019-05, Junghare, Madan, Spiteller, Dieter, Schink, Bernhard
Syntrophorhabdus aromaticivorans is a syntrophically fermenting bacterium that can degrade isophthalate (3-carboxybenzoate). It is a xenobiotic compound which has accumulated in the environment for more than 50 years due to its global industrial usage and can cause negative effects on the environment. Isophthalate degradation by the strictly anaerobic S. aromaticivorans was investigated to advance our understanding of the degradation of xenobiotics introduced into nature, and to identify enzymes that might have ecological significance for bioremediation. Differential proteome analysis of isophthalate- vs benzoate-grown cells revealed over 400 differentially expressed proteins of which only four were unique to isophthalate-grown cells. The isophthalate-induced proteins include a phenylacetate:CoA ligase, a UbiD-like decarboxylase, a UbiX-like flavin prenyltransferase, and a hypothetical protein. These proteins are encoded by genes forming a single gene cluster that putatively codes for anaerobic conversion of isophthalate to benzoyl-CoA. Subsequently, benzoyl-CoA is metabolized by the enzymes of the anaerobic benzoate degradation pathway that were identified in the proteomic analysis. In vitro enzyme assays with cell-free extracts of isophthalate-grown cells indicated that isophthalate is activated to isophthalyl-CoA by an ATP-dependent isophthalate:CoA ligase (IPCL), and subsequently decarboxylated to benzoyl-CoA by a UbiD family isophthalyl-CoA decarboxylase (IPCD) that requires a prenylated flavin mononucleotide (prFMN) cofactor supplied by UbiX to effect decarboxylation. Phylogenetic analysis revealed that IPCD is a novel member of the functionally diverse UbiD family (de)carboxylases. Homologs of the IPCD encoding genes are found in several other bacteria, such as aromatic compound-degrading denitrifiers, marine sulfate-reducers, and methanogenic communities in a terephthalate-degrading reactor. These results suggest that metabolic strategies adapted for degradation of isophthalate and other phthalate are conserved between microorganisms that are involved in the anaerobic degradation of environmentally relevant aromatic compounds.
Syntrophy in Methanogenic Degradation
2018-11-05, Müller, Nicolai, Timmers, Peer, Plugge, Caroline M., Stams, Alfons J. M., Schink, Bernhard
This chapter deals with microbial communities of bacteria and archaea which closely cooperate in methanogenic degradation and perform metabolic functions in this community that neither one of them could carry out alone. The methanogenic degradation of fatty acids, alcohols, most aromatic compounds, amino acids, and others is performed in partnership between fermenting bacteria and methanogenic Archaea. The energy available in these processes is very small, attributing only fractions of an ATP unit per reaction run to every partner. The biochemical strategies taken include in most cases reactions of substrate-level phosphorylation combined with various kinds of reversed electron transport systems in which part of the gained ATP is reinvested into thermodynamically unfavorable electron transport processes. Altogether, these systems represent fascinating examples of energy efficiency at the lowermost energy level that allows microbial life.