Copper inhibited the photoreduction of DCPIP (PS II activity) in a spheroplast preparation of cyanophage AS-1-resistant mutant (An/AS-1). The inhibitory action of Cu2+ was reversed by supplementation with manganese but not with DPC. The results suggest that the site of inhibition was either between the Mn2+ and DPC donation site(s) or close to DPC donation site. Protection of the Hill activity by EDTA further suggestes that Cu2+ interacted with the reaction center.
Three methods for preserving 16 Streptornyces strains were tested: lyophilization on soil and storage in frozen glycerol medium (-20°C), and in liquid nitrogen (-196°C) in the presence of dimethyl sulfoxide. The viability, stability of auxotrophic markers, antibiotic production, and resistance to their own antibiotics for a period up to one year were studied. No variations in the production of and resistance to antibiotics or accumulation of revertants of mutants were evident during the toral preservation time in all three storage methods. A drastic decrease in viable counts was observed after lyophilization on soil. Viability of strains frozen in glycerol and after storage in liquid nitrogen was similar and ranged from 2.3% to 36.6%. Storage of streptomycetes in frozen glycerol is recommended as a quick and reliable method for frequent studies in the laboratory. Storage in liquid nitrogen is recommended as a long-term preservation method.
In this study many yeasts produced heat-stable immunogenic extracellular polysaccharides (EPS). These antigens could be easily detected by competitive enzyme-linked immunosorbent assay (ELISA) or by sandwich ELISA. The antigens of most ascomycetous yeast species tested were almost all species-specific, but those of some basidiomycetous species gave cross reactions with many yeast species, ascomycetes included. Antigens related or similar to those produced by Saccharomyces cerevisiae were detected in culture liquids of 11 strains of the latter, of Kluyveromycesmarxianus, of Zygosaccharomyces florentinus and in several samples of beer and wine, but not in culture liquids of 63 other ascomycetous yeast species, 5 Saccharomyces species included. The EPS ELISA of Saccharomycesexiguus gave positive reactions with culture liquids of Candidamilleri, C. tropicalis, Hanseniaspora vineae, S. dairensis, S. exiguus, S. unisporus and Saccharomycodes ludwigii. The EPS ELISA of Hansenulawickerhamii gave positive reactions with H. henricii, H. wickerhamii and Pichia pinus, thus showing the relationship of these two genera. The EPS ELISA of Geotrichum candidum appeared to be genus-specific like that of filamentous moulds. In addition to several G. candidum strains, G. fermentans, G. klebahnii responded positively, as well as Candida blankii and C. hydrocarbofumarica. The EPS ELISA of Stephanoascus ciferrii was very specific. Except for 10 strains of this species, only Trichosporonterrestre gave a strong positive reaction in the competitive ELISA.
Lipopolysaccharides were isolated from one strain of Leptospirillumferrooxidans and three strains of Thiobacillus ferrooxidans. All strains contained 2-keto-3-deoxyoctonate (KDO), heptose(s) and glucosamine besides neutral sugars, but were devoid of phosphorus. Lipopolysaccharides from iron-grown cells and from sulfur-grown cells had a similar chemical composition. Thus, the chemical properties of the lipopolysaccharide was unaffected by the energy source utilized. Chemical analysis of lipopolysaccharide from Thiobacillus ferrooxidans IFO 14262 revealed the presence of L-rhamnose, D-glucose, L-glycero-D- nianno-heptose, KDO, D-glucosamine, and a lipophilic sugar identified as L-acofriose (3-O-methyl-L-rhamnose). 3-Hydroxymyristic acid was the main fatty acid. By hydrolysis in weak acid, the lipopolysaccharide has been separated into the polysaccharide part ("degraded polysaccharide") and lipid A. Presumably the lipid A contains a glucosamine backbone.
Twelve sludges from plants treating sewage or food industrial waste and ten laboratory sludges acclimated to synthetic sewage were investigated for quinone composition. All tested sludges contained both ubiquinone and menaquinone, and some sludges also produced smaller amounts of demethylmenaquinone and/or rhodoquinone. All the sludges had Q-8 as the most predominant ubiquinone, Q-10 as the second homologue, and Q-9 and other ubiquinones as minor components. The menaquinone profiles of the sludges were considerably variable compared with the ubiquinone composition recorded. The gram-negative aerobic bacteria with the Q-8 quinone system were isolated as the most abundant bacteria from the representative sludges tested, corresponding with the results for ubiquinone profiles of the sludges themselves. There was less correspondence in menaquinone profiles between the bacteria and the sludges from which they were recovered. This study describes the applicability of respiratory quinone profiles as tools for identifying different bacterial populations in activated sludge systems.
Cellular fatty acids, squalene, and sterols in the bacteriochlorophyll-containing bacteria, which include Rhodospirillaceae genera, and Erythrobacter, were investigated. Hydroxy fatty acid composition, and the existence of squalene, Hop-22(29)-ene and Hopan-22-ol explained the heterogeneity of Rhodospirillaceae genera. Erythrobacter longus is distinguished from Rhodospirillaceae species on the basis of hydroxy fatty acid composition. Protomonas extorquens is similar to Rhodopseudomonas palustris and Rhodopseudomonas rutila on the basis of DNA base composition, the quinone system, cellular fatty acid composition, hydroxy fatty acid composition, and occurrence of Hop-22(29)-ene and Hopan-22-ol.
The growth of the strictly anaerobic bacterium Bacteroides ureolyticus was supported by energy production coupled to an anaerobic electron transport from formate or hydrogen to fumarate. There was no growth in a medium containing peptone, yeast extract, and glucose. The growth yield was about 7g dry cells per mol of succinate produced. The electron transport pathway of fumarate reduction was investigated with extracts from cells grown on fumarate plus formate and on fumarate plus hydrogen. Reduction of fumarate to succinate occurred with formate and hydrogen as electron donors, but not with NAD(P)H, lactate, malate, or pyruvate in crude extracts. Activities of formate dehydrogenase, hydrogenase, and fumarate reductase and cytochrome b occurred mainly in a membrane fraction of the cell, c-Type cytochrome was present in both soluble and membrane fractions. All cytochromes were reduced by formate. In the membrane fraction the reduced form of the cytochrome b was immediately oxidized by adding fumarate, and oxidation of the cytochrome c also proceeded, but rather slowly. Apparently the membrane-bound cytochrome b is linked to the reduction of fumarate in B. ureolyticus.
The presence or absence of xylose in the whole cell hydrolysates from all type strains of 41 known species of the genera Bullera, Sporidiobolus, and Sporobolomyces, and three strains of Itersonilia perplexans was examined using high performance liquid chromatography (HPLC) which makes it possible to analyze the carbohydrates without derivatization. Of 16 species of the genus Bullera, 14 had xylose in the cells. The other two species, Bullera salicina and Bullera tsugae, had no xylose in the cells. None of the 25 species of the genera Sporidiobolus and Sporobolomyces had xylose in the cells. The lack of xylose in the cells of seven Sporobolomyces species with Q-9 as the major ubiquinone proves that these species are not yeast phases of certain species of the genus Itersonilia with Q-9, since both the yeast and filamentous phases of Itersonilia perplexans had xylose in the cells. These results show that the presence or absence of xylose in the cells is very useful as one criterion for the generic differentiation of Bullera and Sporobolomyces. The lack of xylose in the cells of B. salicina and B. tsugae indicates that these two species may be closely related to the genus Sporobolomyces. Our HPLC method, by which xylose in the whole cell hydrolysates can be analyzed without derivatization, is a useful way to differentiate taxa by this criterion.