The complete amino acid sequence of ferredoxin from Clostridiumperfringens was established by a combination of Edman degradation and protease digestion. The sequence was: AYKILDTCVSCGACAAECPV-DAISQGDTQFVIDADTCIDCGNCANVCPVGAPVQE. It shows a high degree of similarity to Clostridium pasteurianum ferredoxin (89% homology), and its residual conversions are located at the non-conservative sites of the clostridial-type ferredoxins: at 6 sites, discrete except for the last two, all along the amino(N)-terminal half of the chain, while none at the carboxyl(C)-terminal half (residue numbers from 30 to 55). The occurrence of the 8 cysteine residues at the conservative sites strongly suggested the presence of the conventional two [4Fe-4S] clusters in the molecule.
Ten strains of the Q8-equipped basidiomycetous yeasts classified in the genera Mrakia and Cystofilobasidium were examined for the partial sequence determinations of 18S rRNA and 26S rRNA. The positions determined (in S. cerevisiae) were through 1, 451 to 1, 618 of 18S rRNA and through 493 to 624 and through 1, 601 to 1, 832 of 26S rRNA. The three determinations of the partial sequences of 18S rRNA and 26S rRNA divided the strains examined into two separate groups corresponding to the genera Mrakia and Cystoftlobasidium. Group I contained the strains of M. frigida, M. gelida, M. nivalis, and M. stokesii, and group II contained the strains of C. bisporidii, C. capitatum, and C. infirmominiatum. The inclusion of R. infirmominiatum in the genus Cystofilobasidium as C. infirmominiatum was proved to be phylogenetically reasonable. The data obtained indicate that the division of the Q8-equipped basidiomycetous yeasts into the two genera mentioned above is more natural from the phylogenetic point of view. The utilization of the partial sequences of 18S rRNA and 26S rRNA is discussed, especially in the positions through 1, 451 to 1, 618 of 18S rRNA, for the classification of yeasts (and yeast-like fungi).
On the basis of an extended investigation of morphological, biochemical and physiological properties and its phylogenetic affiliation it is proposed to place Methanomicrobium paynteri (Rivard et al. 1983, 24), validly published in 1984 (Rivard et al. 1984, 25) within a new genus, Methanolacinia gen. nov. as Methanolacinia paynteri comb. nov. Its cell envelope consists of a cytoplasmic membrane and a hexagonally arranged S-layer with a lattice constant of the geometrical subunits of 15.3nm, while the periodic structures on the cell envelope of Methanomicrobium mobile were not characterized. The S-layer contains a glycoprotein subunit with an apparent molecular weight of 155, 000. The polyamine pattern of Methanolacinia paynteri is comparable to that of Methanogenium species but differs significantly from that of Methanomicrobiummobile. Comparative lipid analyses of Methanomicrobium mobile, Methanogenium cariaci, Methanospirillum hungatei and Methanolaciniapaynteri reveal marked differences. Furthermore, Methanolacinia paynteri grows not only on H2/CO2 as reported, but also on 2-propanol/CO2, 2-butanol/CO2 and cyclopentanol/CO2. Together with the low SAB-value of 0.51, which Methanolacinia paynteri shares with Methanomicrobium mobile, these features corroborate the proposed reclassification.
Ammonium assimilation in Saccharolypospora erythrea was mediated by the glutamine synthetase/glutamate synthase pathway depending on the NH4 concentration in the culture medium. Glutamine synthetase formation was repressed by high ammonium concentration, and a rapid loss of its activity occurred after ammonium addition. There was difference in erythromycin biosynthesis and glutamine synthetase levels when L-glutamate of L-glutamine was added as the nitrogen source. When glutamic acid was used as the nitrogen source, the glutamine synthetase activity was high and there was no erythromycin production. But when glutamine was added, the glutamine synthetase level was low and the yield of the antibiotic was high. Glutamate synthase was formed independently of the nitrogen sources used, but glutamate dehydrogenase was present when ammonium was added as a nitrogen source, and it was NADH-dependent. There were high levels of alanine dehydrogenase in cultures grown with glycine, NH4Cl, alanine and glutamine. Alanine most effectively induced the enzyme formation. The results suggest that there was no correlation between the levels of nitrogen-assimilating enzymes and erythromycin biosynthesis in this microorganism.
We describe a simple and rapid procedure for differentiating the koji molds, Aspergillus oryzae and A. sojae, from the closely related species, A. flavus and A. parasiticus. Total DNA from the Aspergillus strains was digested with the restriction enzyme SmaI and was separated by electrophoresis on an agarose gel. The size of the bands, ranging from 3 to 5kb strongly stained with ethidium bromide, was intraspecifically consistent but was interspecifically different. The difference in the size of the bands seemed to be due to the divergence in the nucleotide sequence of nuclear ribosomal DNA. The restriction enzyme cleavage patterns can be readily distinguished on agarose gel, so they will provide a useful tool for identifying koji molds and other related species.
From the cell walls of 45 species (14 genera) of coryneform bacteria, we isolated polysaccharides and teichoic acids covalently linked to the peptidoglycan, and examined their chemical composition. Glycerol teichoic acid appeared in strains of the genera Brevibacterium, Pimelobacter, Rarobacter, in all of the species belonging to the "Arthrobacter nicotianae group" of the genus Arthrobacter, and in A. crystallopoietes belonging to the "A. globiformis group." Strains of the genera Rubrobacter, Aureobacterium, Clavibacter, Brachybacterium, Exiguobacterium and species belonging to the "A. globiformis group" of the genus Arthrobacter contained neutral polysaccharide. Strains of the genera Corynebacterium and Caseobacter had arabinogalactan, as was already known. The sugar composition of the cell-wall polysaccharides in the coryneform bacteria was neither uniform within the genus nor characteristic of each genus in the sugar profile, except for the genera Corynebacterium and Caseobacter which had arabinose and galactose as diagnostic sugars. On the other hand, the kind of cell-wall polymer, whether it is a neutral polysaccharide, teichoic acid or arabinogalactan, was homogeneous within each genus; hence, cell-wall polymers can be used as chemotaxonomic markers for classifying coryneform bacteria.