Biomedical subjects
Publications and source records attributed to O Kandler.
The distribution of the various cell wall and cell envelope (S-layer) polymers among the main lineages of the domain Archaea (Archaebacteria) and the chemical composition and primary structure of polymers forming rigid cell wall sacculi is described. Differences between bacteria and archaea in their sensitivity to antibiotics which inhibit cell wall synthesis in bacteria are discussed.
The cell wall polymer of Natronococcus occultus (DSM 3396) consists of L-glutamate, N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-galacturonic acid, D-glucuronic acid and D-glucose in a molar ratio of 5:7:1:8:0.5:0.3. Partial acid hydrolysis of the cell wall polymer produced soluble fragments that could be separated by HPLC. A gamma-glutamyl dipeptide was isolated. In the intact cell wall polymer, the glutamate residues form a poly-(gamma-glutamine) chain with a length of about 60 monomers, which corresponds to a relative molecular mass of approximately 7700 Da. Two other soluble dimeric fragments, composed of glutamate and either glucosamine or galactosamine in a molar ratio of 1:1, were purified from the hydrolysate, suggesting the presence of two different oligosaccharides linked to the poly-(gamma-glutamine) chain of the intact polymer. The analysis of additional fragments, which were composed of an amino sugar and galacturonic acid or glucose indicated that one oligosaccharide consisted of a glucosamine pentamer in an alpha-1,3 linkage at the reducing end and an oligomer with at least five beta-1,4-linked galacturonic acid residues at the non-reducing end. The second oligosaccharide was comprised of a galactosamine dimer in a beta-1,3 linkage at the reducing end and a maltose unit at the non-reducing end. Both oligosaccharides were linked to the alpha-amide group of the glutamine residues of the poly-(gamma-glutamine) chain. The whole cell wall polymer, which represents a novel type of natural glycoconjugate, has a relative molecular mass of 54 kDa.
The term 'Waldsterben' was introduced in the early 1980s to describe the progressive death of forests that was believed to be occurring in Central Europe as a result of air pollution. Subsequent surveys and investigations have failed to confirm that forests are dying or are even declining over large areas of Central Europe, defined here as consisting of Germany, Switzerland, southeastern France (Alsace), the Czech Republic, northern Italy and Austria. Foliar injury by air pollutants, together with mortality, has occurred, but is generally restricted to specific locations in the Czech Republic and in eastern Germany, such as the Fichtelgebirge. Where foliar damage has been recorded, it can often be attributed to high concentrations of sulphur dioxide, often acting in combination with other stresses (e.g. frost or insects). Outside areas affected by local sources of pollution, there is little, if any, evidence that the crown condition of trees has been adversely affected by pollution over large areas. Instead, climate appears to have a major effect on the crown condition and growth of trees. Measurements and surveys have revealed a very different picture to that forecasted in the mid-1980s. Growth rates of trees and stands in Central Europe are currently higher than have been recorded at any time in the past; the reasons for this are uncertain, although increases in forest area have not substantially contributed to the observed trends. Although declines in individual species in specific areas have been recorded, past records indicate that these do not represent a new phenomenon. Consequently, the terms 'Waldsterben' (forest deaths) and 'neuartige Waldschäden' (novel type of forest damages) should not be used in the context of the phenomenon reported in Central Europe in the 1980s. Instead, different problems should be described separately and the term forest decline used only when there is clear evidence of a general deterioration in the condition of all tree species within a forest.
New insights into genealogic relations among organisms due to similarities of 16S rRNA sequences of more than 1000 representatives of the various groups led to a classification of the presently known living beings into three domains: Bacteria, Archaea (Archaebacteria) and Eucarya. All medical relevant prokaryotes belong to the domain Bacteria. The systematics of the eukaryote microorganisms (protista and fungi) and their numerous medically relevant representatives will be widely changed in the near future due to latest scientific findings on phylogenetic relations. The retrospective analysis of the 16S rRNA phylogenetic tree led to assuming a chemolithoautotrophic origin of life and an early splitting into three domains in an early precellular phase of evolution.
Molecular sequencing technology has brought biology into the era of global (universal) classification. Methodologically and philosophically, global classification differs significantly from traditional, local classification. The need for uniformity requires that higher level taxa be defined on the molecular level in terms of universally homologous functions. A global classification should reflect both principal dimensions of the evolutionary process: genealogical relationship and quality and extent of divergence within a group. The ultimate purpose of a global classification is not simply information storage and retrieval; such a system should also function as an heuristic representation of the evolutionary paradigm that exerts a directing influence on the course of biology. The global system envisioned allows paraphyletic taxa. To retain maximal phylogenetic information in these cases, minor notational amendments in existing taxonomic conventions should be adopted.
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The following putative precursors of the pseudomurein were isolated from trichloroacetic acid extracts of Methanobacterium thermoautotrophicum: a uridine diphosphate activated derivative of glutamic acid and the uridine diphosphate activated peptides (see text). The activated glutamic acid residue and the three activated pepetides lack the glycan components N-acetylglucosamine and N-acetyltalosaminuronic acid present in the intact pseudomurein. In this case uridine diphosphate should be directly linked to the amino group of a glutamic acid residue, which represents a new mode of amino acid and peptide activation.
Molecular structures and sequences are generally more revealing of evolutionary relationships than are classical phenotypes (particularly so among microorganisms). Consequently, the basis for the definition of taxa has progressively shifted from the organismal to the cellular to the molecular level. Molecular comparisons show that life on this planet divides into three primary groupings, commonly known as the eubacteria, the archaebacteria, and the eukaryotes. The three are very dissimilar, the differences that separate them being of a more profound nature than the differences that separate typical kingdoms, such as animals and plants. Unfortunately, neither of the conventionally accepted views of the natural relationships among living systems--i.e., the five-kingdom taxonomy or the eukaryote-prokaryote dichotomy--reflects this primary tripartite division of the living world. To remedy this situation we propose that a formal system of organisms be established in which above the level of kingdom there exists a new taxon called a "domain." Life on this planet would then be seen as comprising three domains, the Bacteria, the Archaea, and the Eucarya, each containing two or more kingdoms. (The Eucarya, for example, contain Animalia, Plantae, Fungi, and a number of others yet to be defined). Although taxonomic structure within the Bacteria and Eucarya is not treated herein, Archaea is formally subdivided into the two kingdoms Euryarchaeota (encompassing the methanogens and their phenotypically diverse relatives) and Crenarchaeota (comprising the relatively tight clustering of extremely thermophilic archaebacteria, whose general phenotype appears to resemble most the ancestral phenotype of the Archaea.
In aqueous trichloroacetic acid extracts of Methanobacterium thermoautotrophicum the following compounds, which are supposed to be precursors in the biosynthesis of the glycan strand of the pseudomurein, were isolated and identified: (i) a disaccharide (compound II) composed of uridine 5'-diphosphate, N-acetylglucosamine and N-acetyltalosaminuronic acid with N-acetylglucosamine at the reducing end, (ii) uridine 5'-diphospho-N-acetylglucosamine, and (iii) uridine 5'-diphospho-N-acetylgalactosamine. However, the corresponding monomeric derivative of N-acetyltalosaminuronic acid could not be detected. It is assumed that N-acetyltalosaminuronic acid may be formed from N-acetylgalactosamine by epimerisation and oxidation at the disaccharide level. These findings indicate that the biosynthetic pathways of murein and pseudomurein are quite different.
The arabinogalactan and peptidoglycan of armadillo-grown Mycobacterium leprae were examined. Within the limits defined by the small amount of material available, the resemblance of these polymers to those of other mycobacteria was confirmed. The polymers were linked by a highly acid-labile bond and the arabinogalactan was itself acid-labile; free arabinose and a variety of oligosaccharides containing both arabinose and galactose, as well as polysaccharide and peptidoglycan, were released by dilute acid. The resonances from anomeric protons in the proton NMR spectrum of the arabinogalactan were similar to those from the arabinogalactan of M. tuberculosis. The composition and structure of the peptidoglycan resembled those of other mycobacteria. The only major difference was the specific replacement of L-alanine by glycine in the peptide of the peptidoglycan.
The immunogenicity and antigenic characteristics of the unique surface layer (S layer) of Methanococcus vanielii was studied with a panel of six monoclonal antibodies. Six surface determinants were identified for the first time, each recognized by one antibody exclusively. The determinants are proteins, located in the S layer, and accessible to antibody in whole, unfixed, as well as formalinized bacteria. Hence the six antigens and antibodies reported here should be useful for rapid identification of new isolates and for taxonomy of methanogens, notably Methanococcaceae. In this connection two novel applications of the slide immunoenzymatic assay were developed for analyses of monoclonal antibodies and their complementary sites in the bacterial envelope.
The term "lactic acid bacteria" is discussed. An overview of the following topics is given: main pathways of homo- and heterofermentation of hexoses, i.e. glycolysis, bifidus pathway, 6-phosphogluconate pathway; uptake and dissimilation of lactose (tagatose pathway); fermentation of pentoses and pentitols; alternative fates of pyruvate, i.e. splitting to formate and acetate, CO2 and acetate or formation of acetoin and diacetyl; lactate oxidation; biochemical basis for the formation of different stereoisomers of lactate.
The antigenic mosaic of the Archaebacterium Methanobacterium thermoautotrophicum, strain delta H, was analyzed with a panel of six monoclonal antibodies. Five antigenic determinants were identified. One contains N-acetyl-D-glucosamine, another contains N-acetyl-D-galactosamine, and a third contains gamma-glutamylalanine (gamma-Glu-Ala). These residues are not involved, at least as immunodominant epitopes, in the other two determinants, one of which contains L-talosaminuronic acid, a component of pseudomurein found only in Methanobacteriaceae. Each of the five determinants was recognized by one monoclonal antibody exclusively. A sixth antibody recognized a structure containing gamma-Glu-Ala that could be either a sixth determinant or a subdeterminant within the site already recognized as containing gamma-Glu-Ala. We postulate that two of the determinants are strain specific, three are species specific, and one is a common antigen.
Biochemical and chemotaxonomical studies were performed on some streptococci from frozen peas and bovine mastitis in an attempt to clarify their taxonomy. The results of the present and earlier studies indicate the pea and mastitis isolates represent two new species of the genus Streptococcus. The isolates from frozen peas are named Streptococcus plantarum sp. nov. and those from mastitis, Streptococcus garvieae sp. nov. The type strains are NCDO 1869 and NCDO 2155, respectively.
After treatment of isolated cells walls of Methanobacterium thermoautotrophicum with sodium hydroxide or anhydrous hydrazine, water soluble glycan strands were obtained. These consisted of alternating (beta 1-3)-linked D-glucosamine and (alpha 1-3)-linked L-talosaminuronic acid residues and their length was about 25 disaccharides. Some of the L-talosaminuronic acid residues remained linked to either glutamic acid or the peptides N-gamma-glutamylalanine and N epsilon-(gamma-glutamylalanyl)lysine, indicating that the peptide moiety of pseudomurein is bound to the carboxyl group of talosaminuronic acid via the amino group of glutamic acid.
Sixty-nine hybridomas were generated to produce monoclonal antibodies to species of methanogens representing three of the four families accepted at the present time: Methanobacteriaceae, Methanococcaceae, and Methanomicrobiaceae. The antibody of each of 29 hybridomas cross-reacted with a methanogen of the same species or genus as the immunizing (homologous) strain, whereas the antibody of the other 40 cell lines reacted only with the homologous strain. Inhibition-blocking experiments with compounds of known composition and structure were used to define the fine specificity of antibodies of four hybridomas representing the four genera of the methanogens used for immunization. The combining site of the monoclonal antibody against M. thermoautotrophicum delta H examined is specific for a structure in the pseudomurein involving N-acetyl-glucosamine but not the (1-3) linkage or the C-terminus gamma-Glu-Ala of the peptide, both of which are characteristic of pseudomurein, the cell-wall peptidoglycan distinctive of the Methanobacteriaceae. In contrast, the monoclonal antibody to the other methanogen of this family examined, M. arboriphilus DH1, recognizes a determinant involving gamma-Glu-Ala. Thus pseudomurein expresses at least two dissimilar antigenic determinants in different portions of the molecule. The monoclonal antibodies against M. vannielii SB and M. hungatei JF1, whose families do not possess pseudomurein, did not display specificity for analogues of pseudomurein.
The determination of the sequence similarity of the ribosomal 16S RNA of many bacteria and a few higher organisms has shown that the methanogenic, halophilic, and acido-thermophilic organisms are phylogenetically separated from the kingdoms of the Eubacteria and Eukaryotes thus representing a third kingdom called "Archaebacteria". Many biochemical and molecular biological features support this conclusion.
The reassociation process of the urea-denatured allosteric L-lactate dehydrogenase from Lactobacillus casei was investigated by hybridization experiments between the reassociating enzymes from L. casei and Lactobacillus curvatus. The quantitatively evaluated hybridization patterns indicate an assembly pathway from the unfolded subunits to the tetrameric state via dimers. The comparison of the kinetics of reassociation and reactivation of the L. casei L-lactate dehydrogenase shows that the tetramer is the only active form.