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Journal ArticleDOI

Clostridium carboxidivorans sp. nov., a solvent-producing clostridium isolated from an agricultural settling lagoon, and reclassification of the acetogen Clostridium scatologenes strain SL1 as Clostridium drakei sp. nov.

TL;DR: A novel solvent-producing, anaerobic clostridium, strain P7(T), was isolated from sediment from an agricultural settling lagoon after enrichment with CO as the substrate and analysis of the 16S rRNA gene sequence showed that it was closely related to Clostridial scatologenes ATCC 25775(T) (99.7% sequence similarity).
Abstract: A novel solvent-producing, anaerobic clostridium, strain P7T, was isolated from sediment from an agricultural settling lagoon after enrichment with CO as the substrate. The metabolism of this Gram-positive, motile, spore-forming rod was primarily acetogenic. Acetate, ethanol, butyrate and butanol were the end-products of metabolism. Strain P7T grew on CO, H2/CO2, glucose, galactose, fructose, xylose, mannose, cellobiose, trehalose, cellulose, starch, pectin, citrate, glycerol, ethanol, propanol, 2-propanol, butanol, glutamate, aspartate, alanine, histidine, asparagine, serine, betaine, choline and syringate as sole substrates. Growth was not supported by methanol, formate, d-arabinose, fucose, lactose, melibiose, amygdalin, gluconate, lactate, malate, arginine, glutamine or vanillate. Nitrate reduction, production of indole, gelatin hydrolysis and aesculin hydrolysis were not observed. Analysis of the 16S rRNA gene sequence of the isolate showed that it was closely related to Clostridium scatologenes ATCC 25775T (99·7 % sequence similarity) and clostridial strain SL1T (99·8 % sequence similarity). Strain SL1 had been classified as a strain of C. scatologenes. However, DNA–DNA reassociation analysis showed that both strain P7T and strain SL1 represented novel clostridial species. It is proposed that strain P7T (=ATCC BAA-624T=DSM 15243T) be classified as the type strain of Clostridium carboxidivorans sp. nov. and that strain SL1T (=ATCC BAA-623T=DSM 12750T) be reclassified as the type strain of Clostridium drakei sp. nov.

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Citations
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TL;DR: This paper critically reviews the existing literature in biomass-derived syngas fermentation into biofuels, specifically, different biocatalysts, factors affecting syngAs fermentation, and mass transfer, and outlines the major challenges of syng as fermentation, key performance index and technology road map, and discusses the further research needs.

533 citations

Journal ArticleDOI
TL;DR: Syngas fermenting microorganisms possess advantageous characteristics for biofuel production and hold potential for future engineering efforts, although genetic tools for such engineering are currently unavailable.

491 citations

Journal ArticleDOI
TL;DR: Tributes are paid to those who discovered acetogens and acetogenesis, and toThose who resolved the acetyl‐CoA pathway to highlight the ecology and physiology of acetogens within the framework of their scientific roots.
Abstract: Acetogens utilize the acetyl-CoA Wood-Ljungdahl pathway as a terminal electron-accepting, energy-conserving, CO(2)-fixing process. The decades of research to resolve the enzymology of this pathway (1) preceded studies demonstrating that acetogens not only harbor a novel CO(2)-fixing pathway, but are also ecologically important, and (2) overshadowed the novel microbiological discoveries of acetogens and acetogenesis. The first acetogen to be isolated, Clostridium aceticum, was reported by Klaas Tammo Wieringa in 1936, but was subsequently lost. The second acetogen to be isolated, Clostridium thermoaceticum, was isolated by Francis Ephraim Fontaine and co-workers in 1942. C. thermoaceticum became the most extensively studied acetogen and was used to resolve the enzymology of the acetyl-CoA pathway in the laboratories of Harland Goff Wood and Lars Gerhard Ljungdahl. Although acetogenesis initially intrigued few scientists, this novel process fostered several scientific milestones, including the first (14)C-tracer studies in biology and the discovery that tungsten is a biologically active metal. The acetyl-CoA pathway is now recognized as a fundamental component of the global carbon cycle and essential to the metabolic potentials of many different prokaryotes. The acetyl-CoA pathway and variants thereof appear to be important to primary production in certain habitats and may have been the first autotrophic process on earth and important to the evolution of life. The purpose of this article is to (1) pay tribute to those who discovered acetogens and acetogenesis, and to those who resolved the acetyl-CoA pathway, and (2) highlight the ecology and physiology of acetogens within the framework of their scientific roots.

468 citations

Journal ArticleDOI
TL;DR: Evidence of 23BD production by three nonpathogenic acetogenic Clostridium species—Clostridia autoethanogenum, C. ljungdahlii, and C. ragsdalei—using carbon monoxide-containing industrial waste gases or syngas as the sole source of carbon and energy is presented.
Abstract: 2,3-Butanediol (23BD) is a high-value chemical usually produced petrochemically but which can also be synthesized by some bacteria. To date, the best microbial 23BD production rates have been observed using pathogenic bacteria in fermentation systems that depend on sugars as the carbon and energy sources for product synthesis. Here we present evidence of 23BD production by three nonpathogenic acetogenic Clostridium species—Clostridium autoethanogenum, C. ljungdahlii, and C. ragsdalei—using carbon monoxide-containing industrial waste gases or syngas as the sole source of carbon and energy. Through an analysis of the C. ljungdahlii genome, the complete pathway from carbon monoxide to 23BD has been proposed. Homologues of the genes involved in this pathway were also confirmed for the other two species investigated. A gene expression study demonstrates a correlation between mRNA accumulation from 23BD biosynthetic genes and the onset of 23BD production, while a broader expression study of Wood-Ljungdahl pathway genes provides a transcription-level view of one of the oldest existing biochemical pathways.

419 citations

Journal ArticleDOI
TL;DR: Pathway engineering to combine established substrate-utilization programs, such as for cellulose, CO2/H2 or CO, with desirable metabolic programs could lead to modular design of strains suitable for many applications.

382 citations

References
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Journal ArticleDOI
TL;DR: In this article, a method was developed to determine submicro amounts of sugars and related substances using a phenol-sulfuric acid reaction, which is useful for the determination of the composition of polysaccharides and their methyl derivatives.
Abstract: Simple sugars, oligosaccharides, polysaccharides, and their derivatives, including the methyl ethers with free or potentially free reducing groups, give an orangeyellow color w-hen treated with phenol and concentrated sulfuric acid. The reaction is sensitive and the color is stable. By use of this phenol-sulfuric acid reaction, a method has been developed to determine submicro amounts of sugars and related substances. In conjunction with paper partition chromatography the method is useful for the determination of the composition of polysaccharides and their methyl derivatives.

45,381 citations


"Clostridium carboxidivorans sp. nov..." refers methods in this paper

  • ...Fructose was measured using the phenol/ sulfuric acid carbohydrate assay (Dubois et al., 1956)....

    [...]

Journal ArticleDOI
TL;DR: The recently‐developed statistical method known as the “bootstrap” can be used to place confidence intervals on phylogenies and shows significant evidence for a group if it is defined by three or more characters.
Abstract: The recently-developed statistical method known as the "bootstrap" can be used to place confidence intervals on phylogenies. It involves resampling points from one's own data, with replacement, to create a series of bootstrap samples of the same size as the original data. Each of these is analyzed, and the variation among the resulting estimates taken to indicate the size of the error involved in making estimates from the original data. In the case of phylogenies, it is argued that the proper method of resampling is to keep all of the original species while sampling characters with replacement, under the assumption that the characters have been independently drawn by the systematist and have evolved independently. Majority-rule consensus trees can be used to construct a phylogeny showing all of the inferred monophyletic groups that occurred in a majority of the bootstrap samples. If a group shows up 95% of the time or more, the evidence for it is taken to be statistically significant. Existing computer programs can be used to analyze different bootstrap samples by using weights on the characters, the weight of a character being how many times it was drawn in bootstrap sampling. When all characters are perfectly compatible, as envisioned by Hennig, bootstrap sampling becomes unnecessary; the bootstrap method would show significant evidence for a group if it is defined by three or more characters.

40,349 citations


"Clostridium carboxidivorans sp. nov..." refers methods in this paper

  • ...A heuristic search was conducted first (using the standard program defaults), followed by a bootstrap analysis (Felsenstein, 1985) to assess the branch points of the resulting phylogenetic trees....

    [...]

Journal Article

16,851 citations

Book
01 May 1989
TL;DR: BCL3 and Sheehy cite Bergey's manual of determinative bacteriology of which systematic bacteriology, first edition, is an expansion.
Abstract: BCL3 and Sheehy cite Bergey's manual of determinative bacteriology of which systematic bacteriology, first edition, is an expansion. With v.4 the set is complete. The volumes cover, roughly, v.1, the Gram-negatives except those in v.3 ($87.95); v.2, the Gram-positives less actinomycetes ($71.95); v.

16,172 citations

Journal ArticleDOI
TL;DR: A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described in this paper.
Abstract: A set of oligonucleotide primers capable of initiating enzymatic amplification (polymerase chain reaction) on a phylogenetically and taxonomically wide range of bacteria is described along with methods for their use and examples. One pair of primers is capable of amplifying nearly full-length 16S ribosomal DNA (rDNA) from many bacterial genera; the additional primers are useful for various exceptional sequences. Methods for purification of amplified material, direct sequencing, cloning, sequencing, and transcription are outlined. An obligate intracellular parasite of bovine erythrocytes, Anaplasma marginale, is used as an example; its 16S rDNA was amplified, cloned, sequenced, and phylogenetically placed. Anaplasmas are related to the genera Rickettsia and Ehrlichia. In addition, 16S rDNAs from several species were readily amplified from material found in lyophilized ampoules from the American Type Culture Collection. By use of this method, the phylogenetic study of extremely fastidious or highly pathogenic bacterial species can be carried out without the need to culture them. In theory, any gene segment for which polymerase chain reaction primer design is possible can be derived from a readily obtainable lyophilized bacterial culture.

10,245 citations