Showing papers in "Archives of Microbiology in 1975"

Characterization of Rhodopseudomonas capsulata.

Abstract: Thirty-three strains of Rhodopseudomonas capsulata have been studied in order to develop a more comprehensive characterization of the species. On the basis of morphological, nutritional, physiological and other properties, the characteristics of an "ideal biotype" have been defined, which can be used to distinguish Rps. capsulata from similar purple bacteria. In this connection, two properties of Rps. capsulata are of particular note: a) sensitivity to penicillin G is 10(3)-10(5) times greater than that shown by closely related species, and b) all strains examined are susceptible to lysis by one or more strains of host species-specific virulent bacteriophages. It appears that members of the species Rps. capsulata form a stringent taxonomic grouping.

Halobacterium volcanii spec. nov., a Dead Sea halobacterium with a moderate salt requirement.

Abstract: A halophilic bacterium was isolated from bottom sediment from the Dead Sea. The organism possessed the properties of the halobacteria, but differed from the known species in two important respects, 1) the cells were disc-shaped and often cupped when grown under optimum conditions, 2) the optimum requirements for sodium chloride was in the range 1.7–2.5 molar which is about half of that generally reported for the halobacteria. The organism was assigned to the genus Halobacterium and described as Halobacterium volcanii spec.nov. The optimum sodium chloride concentration for growth was close to that found in the Dead Sea. The tolerance for magnesium chloride was very high; the organism grew well in media containing magnesium chloride in the concentrations found in the Dead Sea. Halobacterium volcanii is therefore remarkably well fitted for life in the Dead Sea.

Decomposition of specifically 14C-labelled phenols and dehydropolymers of coniferyl alcohol as models for lignin degradation by soft and white rot fungi

Abstract: Several soft and white rot fungi were compared in their ability to degrade specifically 14C-labelled phenols and dehydropolymers of labelled coniferyl alcohol. Furthermore, plant material, which was expected to be specifically labelled in the lignin part was used in the degradation studies. The experiments showed that both groups of fungi were able to release CO2 from methoxyl and carboxyl groups of phenol-carboxylic acids, to degrade side chains of cinnamic acids and cinnamyl alcohols and even to decompose aromatic structures. With the dehydropolymers and the plant material a CO2 release from the methoxyl groups, the side chains and the aromatic carbons was observed. The time dependant course of the CO2 release from these different groups showed in the beginning a higher CO2 evolution from the side chain carbons than from the methoxyl groups, which were later on released to a higher extent. No laccase activity could be detected in the soft rot fungi and the peroxidase activity was lower than in the white rot fungi.

The regulation of carbohydrate metabolism in Klebsiella aerogenes NCTC 418 organisms, growing in chemostat culture.

Abstract: Klebsiella aerogenes NCTC 418 was grown in chemostat cultures (D = 0.17 hr-1; pH 6.8;35 degrees C) that were, successively, carbon-, sulphate-, ammonia-, and phosphate-limited, and which contained as the sole carbon-substrate first glucose, then glycerol, mannitol and lactate. Quantitative analyses of carbon-substrate used and products formed allowed carbon balances to be constructed and direct comparisons to be made of the efficiency of substrate utilzation. With all sixteen cultures, carbon recoveries of better than 90% were obtained. Optimum utilization of the carbon substrate was invariably found with the carbon-limited cultures, the sole products being organisms and carbon dioxide. But the extent to which excess substrate was over-utilized varied markedly with both the nature of the growth-limitation and the identity of the carbon-substrate. In general, sulphate-, ammonia-, and phosphate-limited cultures utilized glycerol more efficiently than mannitol, mannitol better than lactate, and glucose least efficiently. Glucose-containing cultures also synthesized some extracellular polysaccharide. When the carbon source was in excess, a range of acidic compounds generally were excreted. Sulphate-limited cultures, growing on glucose, excreted much pyruvate and acetate, whereas similarly-limited cultures growing on glycerol, mannitol or lactate produced only acetate. Ammonia-limited cultures invariably excreted 2-oxoglutarate and acetate, whereas phosphate-limited cultures produced gluconic acid, 2-ketogluconic acid and acetate, when growing on glucose, but only acetate when growing on mannitol or lactate. From the rates of substrate and oxygen consumption, and the rates of cell synthesis, yield values for both substrate and oxygen were calculated. These showed different trends, but were similar in being highest under carbon-limitation and substantially lower under all other limitations. The physiological significance of these findings, and the probable nature of the regulatory mechanisms underlying "overflow metabolism" are discussed.

Allophycocyanin B (lambdamax 671, 618 nm): a new cyanobacterial phycobiliprotein.

Abstract: A hitherto undescribed red fluorescent phycobiliprotein (maximum emission at ∼ 680 nm), characterized by long wavelength absorption maxima in the visible region at 671 nm (e=172000 M−1·cm−1 per monomer of mol. wt. 30600) and 618 nm, has been purified to homogeneity from a unicellular cyanobacterium, Synechococcus sp., and from a filamentous cyanobacterium, Anabaena variabilis. The name allophycocyanin B has been proposed for the new protein.

A new bacteriochlorophyll from brown-colored Chlorobiaceae

Abstract: A new bacteriochlorophyll has been isolated by thin layer chromatography from all strains of the brown-colored Chlorobiaceae Chlorobium phaeobacteroides and Chlorobium phaeovibriodes. The new bacteriochlorophylle--like the bacteriochlorophylls c and d--represents the major amounts of bacteriochlorophyll a. Bacteriochlorophyll e can be differentiated from the bacteriochlorophylls c and d by its absorption maxima in aceton and its different Rf-value in the thin layer chromatogram. The structure of the new bacteriochlorophyll e has been elucidated on the basis of mass spectra, 1H-and 13C-NMR-spectra, the UV/VIS-spectrum as well as IR-, ORD-, and CD-spectra. The new bacteriochlorophyll has the same relationship to bacteriochlorophyll c as chlorophyll b from green plants to chlorophyll a; therefore, bacteriochlorophyll e represents the first formyl-substituted chlorophyll from bacteria. Similar to the bacteriochlorophylls c and d, the new bacteriochlorophyll e consists of a mixture of at least three homologues which differ from each other by different substituents on the pyrrol rings II and III.

Phosphate utilization and alkaline phosphatase activity in Anacystis nidulans (Synechococcus).

Abstract: Anacystis nidulans (Synechococcus) was maintained in a medium of low phosphate concentration (0.1 mM) and grew with a normal doubling time of 5 hrs at 30°C. Such cultures ahd a normal pigment composition and alkaline phosphatase was detectable at low specific activities only.

Microbial assimilation of hydrocarbons. I. The fine-structure of a hydrocarbon oxidizing Acinetobacter sp.

Abstract: 1. The fine-structure analysis of the hydrocarbon oxidizing microorganism, Acinetobacter sp., demonstrated a cytoplasmic modification resulting from growth on paraffinic and olefinic hydrocarbons. 2. Intracytoplasmic hydrocarbon inclusions were documented by electron microscopy with chemical identifications obtained by gas chromatography and X-ray diffraction. 3. These results demonstrate the ability of a micro-organism to accumulate hydrocarbon substrates intracellularly which, in turn, indicates transport across the cell membrane.

Gene transfer agents, bacteriophages, and bacteriocins of Rhodopseudomonas capsulata.

Abstract: Thirty-three wild type strains of Rhodopseudomonas capsulata were examined for ability to engage in genetic recombination through mediation by “gene transfer agent” (GTA) particles. The genetic exchange assays were based on capacity of strains to produce or receive GTA required for restoration of photosynthetic growth competence to a non-photosynthetic “white” mutant or for acquisition of resistance to rifampicin. A majority of the strains could either produce or receive GTA, and it was demonstrated that the agent is species specific. Possible relations between GTA and bacteriophages or bacteriocins were investigated. Sixteen types of virulent phages active on Rps. capsulata were isolated and their host ranges determined. Tests for transduction by the phages gave uniformly negative results. The viruses showed strict species specificity, but there was no apparent correlation between capacity of the Rps. capsulata strains to donate or receive GTA and susceptibility to the phages. A comparable survey disclosed that most of the bacterial strains were sensitive to or capable of producing bacteriocins; the latter also appear to be unrelated to GTA activity. The collection of bacterial strains was also screened for detection of lysogenic properties. None of the isolates is a “true” lysogen, but phages were detected in cultures of two strains, which may be “phage carriers” or pseudolysogens.

Development of microbodies in candida tropicalis during incubation in a n-alkane medium.

Abstract: Development of microbodies in Candida tropicalis pK 233 was studied mainly by electron microscopical observation. The yeast cells, precultured on malt extract, scarcely contained microbodies and showed very low catalase activity. When the precultured cells were transferred to a n-alkane medium and incubated with shaking, the number of microbodies increased and concomitantly the activity of catalase was enhanced. That is, both the area ratio of microbodies in the cell and the ratio of microbodies to cytoplasm in area increased significantly during the utilization of n-alkanes for 8 hrs. Localization of catalase in the microbodies was demonstrated cytochemically by use of 3,3′-diaminobenzidine, but other organella in the cell, except for vacuoles appearing in the early growth phase and mitochondria, were not stained with this reagent. Microbodies seemed to grow by division. Biogenesis of microbodies in the yeast cells is also discussed.

Bacterial methanogenesis: Acetate as a methane precursor in pure culture

Abstract: Pure cultures of methanogenic bacteria were shown to utilize acetate as a methanogenic substrate. In the presence of hydrogen, both Methanosarcina barkeri and Methanobacterium thermoautotrophicum rapidly converted acetate to methane. This reaction was shown to be dependent on the concentration of hydrogen and acetate. In the absence of hydrogen, acetate was not fermented by methane bacteria. Both the methyl and carboxyl position of acetate were reduced to methane. More 14C-methane was detected from methyl than carboxyl-labeled acetate. The utilization of acetate by cultures of M. thermoautotrophicum was enhanced by addition of CO2/HCO3 −. Methyl labeled acetate was shown to be incorporated into whole cells and converted to 14CO2 and 14CH4 in the presence of H2 and 25 mM CO2/HCO3 −. The importance of acetate utilization in microbial methanogenesis was discussed.

Determination of the Efficiency of Oxidative Phosphorylation in Continuous Cultures of Aerobacter Aerogenes

Abstract: For anaerobic glucose-limited chemostat cultures of Aerobacter aerogenes a values of 14.0 g/mole was found for Y ATP max and a value of 6.8 mmoles ATP/g dry weight/hr for the maintenance coefficient. Both values are much lower than those previously determined for tryptophan-limited anaerobic chemostat cultures. It is concluded that generally the largest part of the maintenance energy is not used for true maintenance processes. For aerobic glucose-limited chemostat cultures two phases could be differentiated. Acetate production started at μ values higher than 0.53. The slopes of the curves relating the specific rates of glucose- and oxygen consumption with μ became higher and lower respectively above the μ value of 0.53. Using the Y ATP values obtained in the anaerobic experiment a P/O ratio of about 1.3 could be calculated for glucose- and tryptophan-limited chemostat cultures. In sulfate-limited chemostat cultures acetate was produced at all growth rates. At high growth rates also pyruvate and α-ketoglutarate were produced. With the Y ATP values obtained in the anaerobic experiment a P/O ratio of about 0.4 was calculated for sulfate-limited chemostat cultures.

Adsorption and selection of rhizobia with ion-exchange papers.

Abstract: Ion exchange papers were used to study the adsorption of 32P-labelled rhizobia on defined surfaces. Two strains of Rhizobium japonicum and one each of R. leguminosarum and R. lupini were compared with Escherichia coli and Bacillus subtilis. The ratio of adsorption to strong and to weak acid papers/strong and weak basic papers was consistantly higher for all rhizobial strains compared to the other bacteria. The process of desorption by increasing the ion-concentration causes about 35% desorption between 0.02 and 0.1 M MgCl2, however, an increase to 1 M does not desorb more labelled Rhizobium japonicum or E. coli cells. The ratio of adsorbed cpm to colony formers, desorbed by 0.1 M NaCl was similar with Rhizobium japonicum for all six ion exchange papers. For E. coli this ratio varied widely for the different papers. The selection of Rhizobium against a more closely related bacterium by this adsorption/desorption procedure was demonstrated with mixed cultures of Rhizobium japonicum and Chromobacterium violaceum giving a more than 80 fold enrichment of the former. Rhizobium japonicum cells, ad/desorbed from all ion exchange papers kept their infectivity and formed nodules on Glycine max with an activity of 20–40 nM C2H4·hr-1·mg nodule-1. A desorption of Rhizobium japonicum from soybean roots also occurred by increasing the ion concentration. 2–3 times as many cells were removed in this way compared to washing with water.

Polybase induced lysis of yeast spheroplasts. A new gentle method for preparation of vacuoles.

Abstract: The polybasic macromolecules DEAE-dextran (diethylaminoethyl-dextran, molecular weight 500000) and poly-DL-lysine (molecular weight 30000-70000) were absorbed with a high affinity by spheroplasts of Candida utilis and subsequently, induced lysis. The extent of lysis of spheroplasts and of the liberated vacuoles was studied under various conditions using alpha-glucosidase activity and soluble arginine as cytoplasmic and vacuolar markers, respectively. Adsorption of polybases was rapidly completed even at 0 degrees C; however, with small doses, lysis was poor at 0-12 degrees C and extensive at temperatures above 12 degrees C. This permitted the completion of adsorption before initiating lysis. The purified vacuoles were also sensitive to polybases though less so than the spheroplasts; however, after lysis of spheroplasts the liberated vacuoles were well protected against the action of polybases. A treatment with polybases which disrupted more than 99% of the spheroplasts left at least 70% of the vacuoles intact. Potassium chloride in high concentrations and calcium chloride in low concentrations inhibited polybase induced lysis of spheroplasts by preventing or even reversing the polybase adsorption. A polyacidic macromolecule, dextran sulfate, could prevent but not reverse the adsorption of polybase and subsequent lysis. Metabolic inhibitors reduced the susceptibility of spheroplasts to polybase induced lysis. Vacuoles isolated from polybase lysed spheroplasts still contained large pools of soluble amino acids, and their ability to transport arginine specifically is a further indication of their functional integrity.

Microbodies in methanol-assimilating yeasts

Abstract: Cells of 3 yeast species capable of assimilating methanol have been examined by electron microscopy. When grown on methanol as the sole source of carbon and energy they contained many microbodies. Cells grown on glucose or ethanol either did not contain such bodies at all, or only to a limited extent.

Cytochemical Localization of Catalase Activity in Methanol-Grown Hansenula polymorpha

Abstract: The localization of peroxidase activity in methanol-grown cells of the yeast Hansenula polymorpha has been studied by a method based on cytochemical staining with diaminobenzidine (DAB). The oxidation product of DAB occurred in microbodies, which characteristically develop during growth on methanol, and in the intracristate space of the mitochondria. The staining of microbodies was H2O2 dependent, appeared to be optimal at pH 10.5, diminished below pH 10 and was inhibited by 20 mM 3-amino 1,2,4 triazole (AT). In contrast to these observations, the reaction in the mitochondria was not H2O2 dependent and not notably affected by differences in pH in the range of 8.5 to 10.5. Microbodies and mitochondria were also stained when H2O2 was replaced by methanol. Appropriate control experiments indicated that in this case methanol oxidase generated the H2O2 for the peroxidative conversion of DAB by catalase. These results suggest that catalase is located in the microbodies of methanol-grown yeasts. A model for a possible physiological function of the microbodies during growth on methanol is put forward.

Lipoic acid content of Escherichia coli and other microorganisms

Abstract: A mutant strain of Escherichia coli K12 requiring lipoic acid, W1485 lip 2 (ATCC 25645), was used to develop a turbidimetric assay for lipoic acid and a polarographic assay based on the oxidation of pyruvate by suspensions of lipoic acid-deficient organisms. The turbidimetric assay was more sensitive with a working range equivalent to 0.2–2.0 ng of dl-α-lipoic acid compared with 5–50 ng for the polarographic method. The mutant responded equally to racemic mixtures of α-lipoic acid, β-lipoic acid and dihydrolipoic acid but gave little response to lipoamide, and other derivatives without prior hydrolysis; 8-methyllipoic acid was a competitive inhibitor of the response to lipoic acid. A high specificity of the mutant for the natural stereoisomer was indicated by the fact that (+)-α-lipoic acid had twice the activity of the racemic mixture. Escherichia coli K12 contained less than 0.05 ng of free (+)-α-lipoic acid per mg dry weight but, depending on the growth substrate, the equivalent of between 13 and 47 ng of (+)-α-lipoic acid per mg dry weight after acid extraction. There was a strong correlation between the lipoic acid content and the sum of the specific activities for the pyruvate and α-ketoglutarate dehydrogenase complexes. Experiments with washed suspensions of Escherichia coli showed only small increases in lipoic acid content (18%) when incubated with pyruvate, cysteine and methionine. When supplied with exogenous lipoic acid the mutant, W1485 lip 2, accumulated very little more than was demanded by its metabolism. The lipoic acid contents of several organisms were measured and correlated with their metabolism.

Conditions for induced fusion of fungal protoplasts in polyethylene glycol solutions.

Abstract: Solutions containing polyethylene glycol MW 6000 (PEG) induced fusion of protoplasts of Penicillium chrysogenum. Balanced heterokaryons were formed by fusion of nutritionally complementing protoplasts. Heterokaryotic fusion products were obtained up to a frequency of 4% of the number of protoplasts, surviving the fusion treatment. Investigation of the conditions, necessary to achieve this high fusion frequency, showed that supplementing the PEG solution with Ca++ and adjustment to high pH gave the best results. Mechanisms of fusion of fungal protoplasts by PEG, calcium and alkaline pH are discussed in view of the obtained results.

Chemical analysis of cell wall regeneration and reversion of protoplasts from Schizophyllum commune.

Abstract: In the presence of MgSO4 as osmotic stabilizer, nucleated protoplasts of Schizophyllum commune developed a large vacuole and could be isolated on the basis of their low buoyant density. All these protoplasts were capable of wall regeneration and about 50 percent reverted to the hyphal mode of growth in liquid medium. The kinetics of the formation of three main cell-wall components, S-glucan (α-1,3-glucan), R-glucan (β-1,3, β-1,6-glucan) and chitin were studied from the onset of regeneration. S-glucan and chitin accumulation as well as RNA and protein synthesis started simultaneously after a short lag, but R-glucan formation was delayed. The reversion to hyphal tubes only began after several hours of rapid R-glucan synthesis. Cycloheximide (0.5 μg/ml), inhibiting protein synthesis by 98% inhibited the formation of R-glucan and the reversion to hyphal growth but the formation of chitin and S-glucan did start and continued seemingly unimpaired for several hours. This indicates that the enzymes responsible for the synthesis of S-glucan and chitin remained intact during protoplast preparation. Polyoxin D inhibited both the synthesis of chitin and R-glucan and also the reversion to hyphal growth. However, the synthesis of S-glucan was not suppressed. These inhibitor studies as well as the kinetics of R-glucan formation during normal regeneration suggest that the synthesis of R-glucan is required for the initiation of hyphal morphogenesis.

Morphology and physiology of Spirochaeta aurantia strains isolated from aquatic habitats.

Abstract: 1. Seven strains of Spirochaeta aurantia were isolated from pond and swamp water by means of a selective technique which utilized the ability of these organisms to move through bacterial filters and to diffuse through agar media. Although most of the isolations were accomplished when enrichment media low in carbohydrates were used, all seven strains were found to be exclusively saccharolytic. 2. The isolates could be divided into two groups on the basis of cell morophology: a loosely coiled group, and a tightly coiled group with markedly smaller wave length and wave amplitude. Spirochetes of the latter group possessed a slightly lower GC content in their DNA. The isolates were facultative anaerobes, synthesized carotenoid pigments which conferred an orange color to aerobic colonies, and utilized a variety of carbohydrates — but not amino acids — as energy sources. Exogenous thiamine was required by six isolates tested. riboflavin by four, and biotin by one. The major products of glucose fermentation were acetate, ethanol, CO2 and H2. Growth of the isolates was inhibited by a variety of antibiotics. Determinations of GC contents of DNA showed that strains of S. aurantia are phylogenetically distant from spirochetes classified in the genera Treponema and Leptospira. 3. S. aurantia populations inoculated in the center of agar medium plates migrated in the form of growth rings toward the periphery of the plates. The rate of migration of glucoseutilizing rings was greatest at low glucose concentrations (e.g., 0.02 g/100 ml). It was concluded that migration of cells present in these rings was mainly due to a chemotactic response to glucose which served both as the attractant and the substrate. Chemotaxis of S. aurantia toward glucose may be used as a selective factor in isolating this bacterium from natural environments. 4. The subspecific epithet stricta is proposed to recognize, taxonomically, the tightly coiled strains of S. aurantia.

CO2 fixation and its regulation in Anacystis nidulans (Synechococcus)

Abstract: Anacystis nidulans (Synechococcus) had a minimal doubling time of 5 hrs at 30 degrees C at saturating light intensity and carbon dioxide concentration. Half maximal growth rates in saturating CO2 occured at a light intensity of 0.54 mW per cm2, and there was an apparent threshold intensity of 0.13 mW per cm2 below which no growth occurred. Growth rate in saturating light was dependent on the concentration of CO2+H2CO3 in the medium, rather than on total dissolved CO2; half maximal rates were estimated at 0.1 mM CO2+H2CO3. Under saturating conditions of light and CO2, 14CO2 was fixed primarily into 3-PGA, and subsequently moved into sugar phosphates and amino acids. Incorporation into aspartate was relatively slow. CO2 fixation was strictly light-dependent. The changes in adenylate and pyridine nucleotide pools were followed in light/dark and dark/light transitions. Whereas adenylates relaxed slowly over 15-20 min to the concentrations characteristic of illuminated cells following the abrupt changes induced by darkening, the sharp drop in intracellular NADPH showed little dark recovery although rapid restoration occurred on reillumination. Other pyridine nucleotides showed no changes during these transitions. The nucleotide specificity and Km of partially purfied GAP dehydrogenase suggest a role for this enzyme in the regulation of CO2 fixation.

Ammonium uptake by nitrogen fixing bacteria I. Azotobacter vinelandii.

Abstract: Both the changes in the activities of nitrogenase, glutamine synthetase and glutamate dehydrogenase and in the extracellular and intracellular NH4+ concentrations were investigated during the transition from an NH4+ free medium to one containing NH4+ ions for a continuous culture of Azotobacter vinelandii. If added in amounts causing 80–100% repression of nitrogenase, ammonium acetate, lactate and phosphate are absorbed completely, whereas chloride, sulfate and citrate are only taken up to about 80%. After about 1–2 hrs the NH4+ remaining in the medium is absorbed too, indicating the induction or activation of a new NH4+ transport system. One of the new permeases allows the uptake of citrate in the presence of sucrose. Addition of inorganic NH4+ salts leads to acidification of the culture. Anaerobiosis suppresses NH4+ transport.

Function of enzymes in wood destroying fungi: II. Multiple forms of laccase in white rot fungi

Abstract: Laccases (p-diphenol-O2-oxidoreductases E.C. 1.14.18.1) of 15 wood rotting fungi were separated by analytical and preparative isoelectric focusing (IEF). Most of the fungi exhibit a pattern of 6–10 extra- and intracellular laccases. A group of laccase bands at pH 3–4 (A complex) and at pH 5–8 (B complex) was common.The laccase isoenzymes showed a qualitative variability with respect to substrate specificities when tested with ortho-, meta- and para-phenolic compounds. There were also differences between intra- and extracellular laccases having identical isoelectric points.This reaction pattern became more complicated when peroxide was added to the enzyme substrates, showing that in some cases it is not possible to make a clear distinction between laccase and peroxidase activity.A quantitative assay of substrate specificities with a polarographic method revealed no gross differences between the two strains tested but a preference for ortho- and paraphenols, whereas meta-phenols are less accessible by laccases.Laccase patterns of 6 geographical races of Pleurotus ostreatus were compared. Inspite of its complexity, there were only slight differences showing their close relationship. In contrast, no comparable agreements have been found between inter-species laccase patterns. The validity of protein or enzyme patterns for taxonomical studies is discussed.Due to the large versatility of the laccase patterns it is concluded that, of the species tested, Polyporus brumalis and Leptoporus litschaueri are best suited for further studies of the laccase function in wood destroying fungi.

Halococcus morrhuae: a sulfated heteropolysaccharide as the structural component of the bacterial cell wall.

Abstract: The qualitative and quantitative composition of purified cell walls of Halococcus morrhuae CCM 859 was determined. Glucose, mannose, galactose; glucuronic and galacturonic acids; glucosamine, galactosamine, gulosaminuronic acid; acetate, glycine and sulfate are found as major constituents. The amino sugars are N-acetylated. It was not possible to fractionate the cell wall in chemically different polymers. Evidence is presented that the major cell wall polymer of this strain is a complex heteroglycan which seems, like the peptidoglycan of most bacteria, to be responsible for the rigidity and stability of the cell wall. In addition it could be proved that this heteroglycan is sulfated and therefore differs considerably from previously described bacterial cell wall polymers.

The Azolla-Anabaena azollae relationship

Abstract: The heterocystous blue-green alga, Anabaena azollae, was isolated from the leaf cavities of the water fern, Azolla caroliniana, where it occurs as an endophyte. The isolated alga was capable of light dependent CO2 fixation and acetylene reduction. Aerobic dark acetylene reduction occurred and was dependent upon endogenous substrates. Vegetative cells of the alga reduced nitro-blue tetrazolium chloride (NBT) to blue formazan. Heterocysts did not. Heterocysts reduced triphenyl tetrazolium chloride (TTC) to red formazan faster than vegetative cells. Reduction of TTC by both heterocysts and vegetative cells was much more rapid than has been reported for free-living heterocystous blue-green algae. Both NBT and TTC inhibited acetylene reduction and CO2 fixation. The inhibition by TTC was more closely correlated to the time of exposure of the cells to the reagent and to the amount of deposition per cell than to the number of cells containing red formazan. No differential inhibition of acetylene reduction versus CO2 fixation was observed. Autoradiography showed that CO2 fixation occurred only in vegetative cells. Heterocysts caused a darkening of nuclear emulsions (chemography). This observation has been employed by others as an index of reducing activity in these cells. DCMU inhibited the acetylene reducing capacity of alga isolated from dark pretreated fronds more rapidly and to a greater extent than that in alga isolated from light pretreated fronds. Ammonia in excess of 5 mM was required before any inhibition of acetylene reduction was observed under either aerobic or anaerobic conditions in the light.

Effect of vanadium on growth, chlorophyll formation and iron metabolism in unicellular green algae.

Abstract: In the presence of vanadium, growth of Scenedesmus obliquus and Chlorella pyrenoidosa was increased five to sixfold as determined by dry weight, when cultured under autotrophic conditions for 7 days. The stimulation by vanadium decreased with increasing stability towards hydrolysis of the iron(III)-compounds added. Pentavalent vanadium (20 mug V/1 as NH4VO3) was able to overcome completely a limited iron-deficiency in the algae following growth in presence of 1.8 - 10(-5) m ferric chloride. Vanadium did not alter the iron uptake into the algal cells. 90% of offered 48V was taken up by Scenedesmus obliquus during 5 days of growth, and 21% thereof were found in the chloroplast fraction. In presence of vanadium, the chlorophyll formation was stimulated in Scenedesmus obliquus. This stimulation by vanadium was found to be light-dependent but occurred to a certain extent in the dark also. The main porphyrin of the yellow mutant 211-11h/20 of Chlorella vulgaris was identified as protoporphyrin-IX. The formation of this compound was stimulated by vanadium within 10 days up to 83%. The role of vanadium in the biosynthesis of chlorophylls is discussed.

Microbial assimilation of hydrocarbons. II. Intracytoplasmic membrane induction in Acinetobacter sp.

Abstract: 1. The induction of intracytoplasmic membranes was demonstrated to occur in Acinetobacter sp. when grown on hexadecane, heptadecane, and hexadec-1-ene. 2. Evidence for a physical relationship between the cytoplasmic hydrocarbon "pools" and the intracytoplasmic membranes is presented. 3. The specificity of cytoplasmic pooling of hydrocarbons and the induction of intracytoplasmic membranes was investigated in relationship to hydrocarbon oxidation. 4. These results suggests that both processes are required for the growth of Acinetobacter sp. on hydrocarbons.

Specific inhibitors of methane oxidation in Methylosinus trichosporium

Abstract: Methane oxidation by washed cell suspensions of Methylosinus trichosporium OB3B was selectively inhibited by 25 compounds, including metal binding components such as carbon monoxide (85% O2: 15% CO), KCN (10-6 M), αα′-dipyridyl (10-4 M), 8-quinolinol (10-4 M), thiosemicarbazide (10-5 M), thiourea (10-5 M), hydroxylamine (10-4 M), histidine (10-2 M), British Anti-Lewisite (5x10-3 M), and miscellaneous known inhibitors of other oxygenases. A role for copper in the methane oxygenase system was suggested by the pattern of inhibition and relief of inhibition by added metal ions.

Catabolism of D-fructose and D-ribose by Pseudomonas doudoroffii. I. Physiological studies and mutant analysis.

Abstract: Pseudomonas doudoroffii, a strict aerobe of marine origin, was able to utilize fructose and ribose but not glucose, gluconate, or other hexoses, pentoses, or sugar alcohols as sole sources of carbon and energy Evidence was presented indicating that in this organism fructose was utilized via an inducible P-enolpyruvate: fructose phosphotransferase system (FPTS) which catalyzed the phosphorylation of fructose in the 1 position The resulting fructose-1-P (F-1-P) was converted to fructose-1,6-P2 (FDP) by means of an inducible 1-P-fructokinase (1-PFK) The subsequent conversion of FDP to pyruvate involved enzymes of the Embden-Meyerhof pathway (EMP) which, with the exception of glyceraldehyde-3-P dehydrogenase (G3PDH), were constitutive Two G3PDH activities were detected, one of which was inducible and NAD-dependent while the other was constitutive and NADP-dependent Cell-free extracts of P doudoroffii also contained enzymes of the methylglyoxal pathway (MGP) which converted dihydroxyacetone-P to pyruvate The low specific activities of enzymes of this pathway as compared to the EMP suggested that the major route of FDP catabolism was via the latter pathway 2 Ribose catabolism appeared to involve an inducible uptake system and an inducible ribokinase, the resulting ribose-5-P being converted to glyceraldehyde-3-P and fructose-6-P (F-6-P) by means of constitutive activities of the pentose-P pathway The F-6-P formed as a result of these reactions was converted to FDP by means of a constitutive 6-P-fructokinase (6-PFK) Since no activity converting fructose or F-1-P to F-6-P could be detected in cell-free extracts of P doudoroffii, the results suggested that fructose and ribose were catabolized via 1-PFK and 6-PFK, respectively, the two pathways converging at the level of FDP Further evidence for this suggestion was obtained from a mutant which lacked an NAD-dependent G3PDH, accumulated FDP from both fructose and ribose, and was not able to grow on either of these compounds 3 Ribose grown cells had increased amounts of the fructose uptake system and 1-PFK suggesting that a compound (or compounds) common to the catabolism of both fructose and ribose acted as the inducer(s) of these activities Evidence was presented suggesting that the probable inducer(s) of 1-PFK and FPTS could be FDP, glyceraldehyde-3-P, or dihydroxyacetone-P 4 A mutant unable to grow on fructose was characterized and found to lack FPTS while retaining 1-PFK and other enzyme activities of the EMP and MGP, indicating that a functional FPTS was essential for growth on fructose and suggesting that all or most of this sugar was catabolized via F-1-P