Showing papers in "Archives of Microbiology in 1980"

Characterization of an acetate-decarboxylating, non-hydrogen-oxidizing methane bacterium.

Abstract: A methanogenic bacterium, commonly seen in digested sludge and referred to as the "fat rod" or Methanobacterium soehngenii, has been enriched to a monoculture and is characterized. Cells are gramnegative, non-motile and appear as straight rods with flat ends. They form filaments which can grow to great lengths. The structure of the outer cell envelop is similar to Methanospirillum hungatii. The organism grows on a mineral salt medium with acetate as the only organic component. Acetate is the energy source, and methane is formed exclusively from the methyl group. Acetate and carbon dioxide act as sole carbon source and are assimilated in a molar ratio of about 1.9:1. The reducing equivalents necessary to build biomass from these two precursors are obtained from the total oxidation of some acetate. Hydrogen is not used for methane formation and is not needed for growth. Formate is cleaved into hydrogen and carbon dioxide. Coenzyme M was found to be present at levels of 0.35 nmol per mg of dry cells and F420 amounted to 0.55 microgram per mg protein. The mean generation time was 9 days at 33 degrees C.

The Sulfolobus-“Caldariella” group: Taxonomy on the basis of the structure of DNA-dependent RNA polymerases

Abstract: The similarity of the morphology and of DNA composition, the homology of the component patterns of DNA-dependent RNA polymerases and their immunochemical crossreactivity support the conclusion that several extreme thermoacidophiles are related to each other. We name two new species of the genus Sulfolobus. The first, Sulfolobus solfataricus (DSM 1616 and DSM 1617) has the same GC content in its DNA and the same general properties as S. acidocaldarius, but differs significantly from the latter species in the molecular weights of the 11 components of its RNA polymerase and in the salt requirements of this enzyme. The second, Sulfolobus brierleyi, DSM 1651, differs from S. acidocaldarius in several respects. The cells show much less stability at neutral pH. The GC content is significantly lower. The RNA polymerase lacks two components present in the enzymes from the other species. The residual 9 components show larger size differences from the homologous subunits of the S. acidocaldarius enzyme. Like the enzyme from S. solfataricus, the polymerase from S. brierleyi yields an incomplete immunochemical crossreaction with an antibody against the RNA polymerase from S. acidocaldarius. The isolates DSM 1616 and DSM 1617 of Sulfolobus solfataricus are probably identical with or similar to the “Caldariella” strains MT 3 and MT 4, isolated by de Rosa et al. (1975). Like all other known archaebacterial RNA polymerases the enzymes from these species are insensitive to rifampicin and streptolydigin.

Growth parameters ( K s , μ max , Y s ) of Methanobacterium thermoautotrophicum

Abstract: Methanobacterium thermoautotrophicum was grown on a mineral salts medium in a fermenter gassed with H2 and CO2, which were the sole carbon and energy sources. Under the conditions used the bacterium grew exponentially. The dependence of the growth rate (μ) on the concentration of H2 and CO2 in the incoming gas and the dependence of the growth yield ( $$Y_{CH_4 }$$ ) on the growth rate were determined at pH 7 (the pH optimum) and 65° C (the temperature optimum). The curves relating growth rate to the H2 and CO2 concentration were hyperbolic. From reciprocal plots apparent K s values for H2 and CO2 and μmax were obtained: app. $$K_{{\text{H}}_{\text{2}} }$$ = 20%; app. $$K_{{\text{CO}}_{\text{2}} }$$ = 11%; μ = 0.69 h-1; t δ (max)=1 h. $$Y_{CH_4 }$$ was 1.6 g mol-1 and almost independent of the growth rate, when the rate of methane formation was not limited by the supply of either H2 or CO2. The yield increased to near 3 g mol-1 when H2 or CO2 were limiting. These findings indicate that methane formation and growth are less tightly coupled at high concentrations of H2 or CO2 in the medium than at low concentrations. The physiological significance of these findings is discussed.

C-phycocyanin as a storage protein in the blue-green alga Spirulina platensis

Abstract: The possibility that c-phycocyanin serves as a nitrogen source in Spirulina platensis during nitrogen starvation was studied. The following evidence was obtained in support of this idea. 1. Under favourable conditions for growth, c-phycocyanin existed in large excess in the algal cells. 2. When the supply of nitrogen was low, about 30–50% of the c-phycocyanin disappeared without any effect on the maximal growth rate. 3. A culture which was deprived of nitrogen continued to grow unaffectedly for a period, the duration of which depended on the c-phycocyanin content in the cell before nitrogen starvation was initiated. 4. c-phycocyanin was the only nitrogenous compound that was depleted during the course of nitrogen starvation when growth was yet unaffected. 5. When protein synthesis was inhibited either by nitrogen starvation or by methionine sulfoximine (MSO), phycocyanin content began to decline immediately and growth continued at normal rates as long as c-phycocyanin did not decline below 50%. 6. The decrease in c-phycocyanin content during nitrogen starvation was accompanied by an increase in proteolytic activity.

Dynamic aspects of phycobilisome structure

Abstract: In exponentially growing cells of Synechococcus sp. 6301, over 95% of the phycobiliproteins are located in phycobilisomes, and the remainder is present in the form of low molecular weight aggregates. In addition to the subunits of the phycobiliproteins (C-phycocyanin, allophycocyanin, allophycocyanin B), the phycobilisomes of this unicellular cyanobacterium contain five non-pigmented polypeptides. During the initial phase of starvation (24 h after removal of combined nitrogen from the growth medium), the phycobiliproteins in the low molecular weight fraction largely disappeared. Phycocyanin was lost more rapidly from this fraction than allophycocyanin. Simultaneous changes in the phycobilisome were (1) a decrease in sedimentation coefficient, (2) a decrease in phycocyanin: allophycocyanin ratio, (3) a shift in the fluorescence emission maximum from 673 to 676 nm, and (4) a selective complete loss of a 30,000 dalton non-pigmented polypeptide. Upon extensive nitrogen starvation (72 h), the intracellular level of phycocyanin decreased by over 30-fold. These results indicate that in the early stage of nitrogen starvation, the free phycobiliproteins of the cell are degraded, as well as a significant proportion of the phycocyanin from the periphery of the phycobilisome. However, the structures partially depleted of phycocyanin still function efficiently in energy transfer. On extended starvation, total degradation of residual phycobilisomes takes place, possibly in conjunction with the detachment of these structures from the thylakoids. None of the effects of the absence of combined nitrogen were seen when cells were starved in the presence of chloramphenicol, or in a methionine auxotroph starved for methionine.

Treponema bryantii sp. nov., a rumen spirochete that interacts with cellulolytic bacteria

Abstract: A saccharolytic spirochete that associated and interacted with cellulolytic bacteria was isolated from bovine rumen fluid. Isolation was accomplished by means of a procedure involving serial dilution of a sample of rumen fluid into a cellulose-containing agar medium. Clear zones appeared within the medium as a result of cellulose hydrolysis by rumen bacteria. The saccharolytic spirochete and a cellulolytic bacterium later identified as a strain of Bacteroides succinogenes were isolated from the clear zones. The spirochete did not utilize cellulose, but grew in coculture with the cellulolytic bacterium in cellulose-containing media. When cocultured in these media the spirochete used, as fermentable substrates, soluble sugars released from cellulose by the cellulolytic bacterium. In cellulosecontaining agar medium the spirochete enhanced cellulose breakdown by the B. succinogenes strain. Electron microscopy showed that the helical spirochete cells possessed an outer sheath, a protoplasmic cylinder, and two periplasmic fibrils. Under a CO2 atmosphere, in a reduced medium containing inorganic salts, rumen fluid, glucose, and NaHCO3, the spirochete grew to a final density of 1.9×109 cells/ml. Succinate, acetate, and formate were products of the fermentation of glucose by growing cells. CO2 (HCO3 -), branched short-chain fatty acids, folic acid, biotin, niacinamide, thiamine, pyridoxal, and a carbohydrate were required for growth of the spirochete. The results of this study indicated that the rumen spirochete represents a new species of Treponema. It is proposed that the new species be named Treponema bryantii.

A comparative study on the composition of chlorosomes (Chlorobium vesicles) and cytoplasmic membranes from Chloroflexus aurantiacus strain Ok-70-fl and Chlorobium limicola f. thiosulfatophilum strain 6230

Abstract: Highly purified fractions of chlorosomes and cytoplasmic membranes were isolated from Chloroflexus aurantiacus Ok-70-fl and Chlorobium limicola 6230. These fractions were comparatively analyzed for their pigmentation, phospholipid, glycolipid, and cytochrome c content as well as for their specific activities of succinate dehydrogenase and NADH-oxidase. The data showed that there are some differences in pigmentation and phospholipid content between the isolated fractions of Chloroflexus and Chlorobium. Chlorosomes of Chloroflexus contained a specific BChl a-complex with a characteristic absorption maximum at about 790 nm. This BChl a-complex could not be detected in spectra of chlorosomes from Chlorobium. The near infrared region of the spectra of the isolated cytoplasmic membranes of both organisms revealed considerable differences: The BChl a-complexes of Chloroflexus membranes exhibited peaks at 806 and 868 nm whereas the membranes of Chlorobium had a single BChl a-peak at 710 nm. In contrast to the findings with Chlorobium the chlorosomes of Chloroflexus contained at least twice as much phospholipids as did the cytoplasmic membranes. In Chlorobium the phospholipid content of cytoplasmic membranes is three times that of their chlorosomes. The distribution of all other components (carotenoid composition, enzyme activities, cytochrome c content, and glycolipids) was about the same in both strains. From the data it was concluded that differences in the organization of the photosynthetic apparatus are mainly based on differences of the organization of the photosynthetic units in the cytoplasmic membrane and probably the kind of linkage of the light harvesting system in the chlorosomes with the reaction center in the cytoplasmic membranes.

Methanol Metabolism in Yeasts: Regulation of the Synthesis of Catabolic Enzymes

Abstract: The regulation of the synthesis of four dissimilatory enzymes involved in methanol metabolism, namely alcohol oxidase, formaldehyde dehydrogenase, formate dehydrogenase and catalase was investigated in the yeasts Hansenula polymorpha and Kloeckera sp. 2201. Enzyme profiles in cell-free extracts of the two organisms grown under glucose limitation at various dilution rates, suggested that the synthesis of these enzymes is controlled by derepression — represion rather than by induction — repression. Except for alcohol oxidase, the extent to which catabolite repression of the catabolic enzymes was relieved at low dilution rates was similar in both organisms. In Hansenula polymorpha the level of alcohol oxidase in the cells gradually increased with decreasing dilution rate, whilst in Kloeckera sp. 2201 derepression of alcohol oxidase synthesis was only observed at dilution rates below 0.10 h−1 and occurred to a much smaller extent than in Hansenula polymorpha.

Capacity of chromatiaceae for chemotrophic growth. Specific respiration rates of Thiocystis violacea and Chromatium vinosum

Abstract: The capacity for chemoautotrophic, mixotrophic and organotrophic growth in the dark was tested with 45 strains of 17 species (11 genera) of the Chromatiaceae. The auxanographic deep agar shake culture method was used; the gas phase contained 5% O2 and 1% CO2 in N2. All strains tested of Chromatium vinosum, C. minus, C. violascens, C. gracile, Thiocystis violacea, Amoebobacter roseus, Thiocapsa roseopersicina gave positive growth responses under chemoautotrophic and mixotrophic conditions (extra carbon source acetate); one strain of Thiocapsa roseopersicina grew also organotrophically on acetate alone. No growth was obtained with the remaining 17 strains of ten species. None of the five type species (three genera) of the Chlorobiaceae grew under chemotrophic conditions. With Thiocystis violacea 2311 a growth yield of 11.3g dry weight per mol thiosulfate consumed was obtained under chemoautotrophic conditions; under mixotrophic conditions with acetate the yield increased to 69g dry weight per mol thiosulfate consumed. With Thiocystis violacea 2311 maximal specific respiration rates were obtained with thiosulfate as electron donor irrespective of the presence or absence of sulfur globules in the cells; organic substrates served as carbon sources only and did not support respiration. With Chromatium vinosum D utilization of thiosulfate was not constitutive; maximal respiration rates on thiosulfate were obtained only with thiosulfate grown cells containing sulfur globules. Respiration rates were further increased by malate, fumarate or propionate; these substrates also served as sole electron donors for respiration. Acetate and pyruvate were used as carbon sources only. The ecological significance of the chemotrophic metabolism is discussed.

Nickel, a component of factor F430 from Methanobacterium thermoautotrophicum

Abstract: Methanobacterium thermoautotrophicum, growing on medium supplemented with 2 μmol 63NiCl2/l, was found to take up 1.2 μmol 63Ni per g cells (dry weight). More than 70% of the radioisotope was incorporated into a compound, which dissociated from the protein fraction after heat treatment, was soluble in 70% acetone, and could be purified by chromatography on QAE-Sephadex A-25, Sephadex G-25, and DEAE cellulose. The purified 63Ni labelled compound had an absorption spectrum and properties identical to those of factor F430 and is therefore considered to be identical with factor F430.

Autotrophic CO 2 fixation in Chlorobium limicola . Evidence for the operation of a reductive tricarboxylic acid cycle in growing cells

Abstract: Chlorobium limicola was grown on a mineral salts medium with CO2 as the main carbon source supplemented with specifically labeled 14C propionate and the incorporation of 14C into alanine (≙ intracellular pyruvate), aspartate (≙ oxaloacetate), and glutamate (≙ α-ketoglutarate) was studied in long term labeling experiments. During growth in presence of propionate 30% of the cell carbon were derived from propionate and 70% from CO2. Propionate was not oxidized to CO2. All three amino acids were found to be labeled. The labeling patterns indicate that propionate was assimilated via propionyl CoA, methylmalonyl CoA and succinyl CoA. When 1-14C propionate was the labeled precursor no radioactivity was found in the carboxyl group(s) of alanine, aspartate and glutamate, excluding the incorporation of propionate into the amino acids via succinate oxidation to fumarate. With 1-14C propionate preferentially aspartate (C-3) and glutamate (C-2) became labeled, with 2-14C propionate alanine (C-3) and glutamate (C-4). These findings indicate that propionate was incorporated into the amino acids via succinyl CoA, α-ketoglutarate, isocitrate, and citrate, followed by a si-type cleavage of citrate to oxaloacetate and acetyl CoA (or acetate). Similar experiments with U-14C acetate confirm these conclusions. Thus, all reactions of the proposed reductive tricarboxylic acid cycle could be demonstrated in autotrophically growing cells.

Short-term ammonium inhibition of nitrate utilization by Anacystis nidulans and other cyanobacteria

Abstract: Ammonium at low concentrations caused a rapid and effective inhibition of nitrate utilization in the light by the cyanobacterium Anacystis nidulans without affecting the cellular level of nitrate reductase activity. The inhibition was reversible, and the ability of the cells to utilize nitrate was restored immediately after ammonium had been exhausted. The inhibitory effect was dependent on consumption by the cells of the added ammonium which was rapidly incorporated into amino acids. In the presence of L-methionine-d,l-sulfoximine (MSX) or azaserine, inhibitors of the glutamine synthetase-glutamate synthase pathway, ammonium did not exhibit any inhibitory effect on nitrate utilization. Ammonium assimilation, rather than ammonium itself, seems to regulate nitrate utilization in A. nidulans. Short-term inhibition by ammonium of nitrate utilization and its prevention by MSX were also demonstrated in the filamentous cyanobacteria Anabaena and Nostoc.

The effect of nisin on murein synthesis.

Abstract: Nisin inhibits murein synthesis with concomitant accumulation of undecaprenyl-pyrophospho-MurNAc(pentapeptide) (lipid intermediate I). This inhibition is caused by the formation of a complex between the antibiotic and lipid intermediate I. Undecaprenyl-pyrophospho-MurNAc(pentapeptide)-GlcNAc (lipid intermediate II) also forms a complex with nisin. However, when murein synthesis is inhibited by nisin, this latter complex is not formed since lipid intermediate II is no longer synthesized.

Anaerobic oxidation of thiosulfate and elemental sulfur in Thiobacillus denitrificans

Abstract: Thiobacillus denitrificans strain RT could be grown anaerobically in batch culture on thiosulfate but not on other reduced sulfur compounds like sulfide, elemental sulfur, thiocyanate, polythionates or sulfite. During growth on thiosulfate the assimilated cell sulfur was derived totally from the outer or sulfane sulfur. Thiosulfate oxidation started with a rhodanese type cleavage between sulfane and sulfone sulfur leading to elemental sulfur and sulfite. As long as thiosulfate was present elemental sulfur was transiently accumulated within the cells in a form that could be shown to be more reactive than elemental sulfur present in a hydrophilic sulfur sol, however, less reactive than sulfane sulfur of polythionates or organic and inorganic polysulfides. When thiosulfate had been completely consumed, intracellular elemental sulfur was rapidly oxidized to sulfate with a specific rate of 45 natom S°/min·mg protein. Extracellularly offered elemental sulfur was not oxidized under anaerobic conditions.

Vectorial electron transport in Degulfovibrio vulgaris (Marburg) growing on hydrogen plus sulfate as sole energy source

Abstract: Desulfovibrio vulgaris (Marburg) was grown on hydrogen plus sulfate as sole energy source in a medium containing excess iron. The topography of electron transport components was investigated. The bacterium contained per mg cells (dry weight) 30U hydrogenase (1U=1 μmol/min), 35 μg desulfoviridin (= bisulfite reductase), 0.6 U adenosine phosphosulfate reductase, 30 mU thiosulfate reductase, 0.3 nmol cytochrome c3 (Mr=13,000), 0.04 nmol cytochrome b, 0.85 nmol menaquinone, and 0.4 nmol ferredoxin. Hydrogenase (>95%) and cytochrome c3 (82%) were localized on the periplasmic side and desulfoviridin (≈95%), adenosine phosphosulfate reductase (87%), thiosulfate reductase (74%), and ferredoxin (71%) on the cytoplasmic side of the cytoplasmic membrane; menaquinone and cytochrome b were exlusively found in the membrane fraction. The location of the oxidoreductases indicate that in D. vulgaris (Marburg) H2 oxidation and sulfate reduction take place on opposite sides of the cytoplasmic membrane rather than on the same side, as has recently been proposed.

Polyhydric alcohol production and intracellular amino acid pool in relation to halotolerance of the yeast Debaryomyces hansenii

Abstract: Changes in polyol production and the intracellular amino acid pool were followed during the growth cycle of Debaryomyces hansenii in 4 mM and 2.7 M NaCl media. The intracellular levels of polyols were markedly enhanced by high salinity, the dominant solutes being glycerol in log phase cells and arabinitol in stationary phase cells. At low salinity arabinitol was the most prominent intracellular solute throughout the growth cycle. There were no major changes in the composition of the total amino acid pool with changes in cultural salinity. The amount of total free amino acids related to cell dry weight was 15–50% lower in cells cultured in 2.7 M NaCl as compared to 4 mM NaCl media.

Vanillic acid metabolism by the white-rot fungus Sporotrichum pulverulentum

Abstract: Vanillic acid metabolism was studied in wild-type Sporotrichum pulverulentum and three different mutants. Vanillic acid was found to be oxidatively decarboxylated to methoxyhydroquinone (MHQ) and simultaneously reduced to vanillin and vanillyl alcohol to different degrees depending upon the cultivation conditions. The reducing pathway cannot be utilized unless the fungus has access to an easily metabolized carbon source such as glucose or cellobiose, while decarboxylation takes place in cultures with only vanillic acid present. Polymerization reactions also occurred in the culture solutions. Some evidence for reoxidation of vanillin and vanillyl alcohol was obtained in vivo, and in vitro experiments using horseradish peroxidase.

NADH as electron donor for the photosynthetic membrane of Chlamydomonas reinhardii

Abstract: A chloroplast fraction from Chlamydomonas reinhardii cells can oxidize NADH in the light, unlike chloroplasts of higher plants. The Chlamydomonas preparation catalyzes electron flow from NADH to methylviologen or ferredoxin to evolve hydrogen (in the presence of a hydrogenase) or take up oxygen. The NADH photooxidation is sensitive to rotenone, dibromothymoquinone and dicyclohexylcarbodiimide. This suggests that a rotenone sensitive NADH dehydrogenase is coupled on the plastoquinone reduction site of the potosynthetic electron flow system. On sonication of the particles NADH photooxidation is lost but may be restored by a protein fraction from an acetone extract plus plastocyanin.

Relations between d-ribulose-1,5-bis-phosphate carboxylase, carboxysomes and co2 fixing capacity in the obligate chemolithotroph thiobacillus-neapolitanus grown under different limitations in the chemostat

Abstract: An adaptation of the d-ribulose-1,5-bisphosphate carboxylase (RuBPCase) activity to changing CO2 concentrations in the growth medium in the chemostat was observed in the obligate chemolithotroph Thiobacillus neapolitanus. RuBPCase activity has been separated in a soluble and particulate fraction. The activity of the particulate fraction appeared to be associated with the carboxysomes.

Excretion of a protease by Serratia marcescens

Abstract: Excretion of an extracellular protease of Serratia marcescens ATCC 25419 occurred during logarithmic growth and was highest (per cell) when cultures reached the stationary growth phase. Production of the extracellular protease was induced by leucine or casein in minimal medium or by growth in tryptone-yeast medium. In the late stationary phase an intracellular protease activity accumulated which was also observed in mutants with very low extracellular protease activity. The excreted protease was the dominant protein in the growth medium. The protease was purified to homogeneity by column chromatography on Bio-Gel P-100 and on DEAE-cellulose. Quantitative amino acid analysis revealed the absence of sulfur-containing amino acids. The enzyme consists of one polypeptide chain. A molecular weiht of 51,000 and 55,000 was estimated using polyacrylamide gel electrophoresis and chromatography on Bio-Gel P-100 respectively. The enzyme cleaved only N-alpha-benzoyl-DL-lysine-and-arginine-nitroanilides but not the corresponding leucine or tyrosine derivatives nor a set of di- and tripeptides.

Influence of manganese on morphology and cell wall composition of Aspergillus niger during citric acid fermentation.

Abstract: Morphology and cell wall composition of Aspergillus niger were studied under conditions of manganese sufficient or deficient cultivation in an otherwise citric acid producing medium. Omission of Mn2+ (less than 10-7 M) from the nutrient medium of Aspergillus niger results in abnormal morphological development which is characterized by increased spore swelling, and squat, bulbeous hyphae. Fractionation and analysis of manganese deficient cell walls revealed increased chitin and reduced β-glucan contents as well as reduction of galactose containing polymers, as compared to cell walls from manganese sufficient grown hyphae. Addition of copper induced the same effect as manganese deficiency, both on morphology and cell wall composition. Addition of cycloheximide also produced a very similar type of morphology with increased chitin and reduced β-glucan contents of the cell wall but its effect on galactose was less pronounced.

Methane formation from fructose by syntrophic associations of Acetobacterium woodii and different strains of methanogens.

Abstract: When Acetobacterium woodii was co-cultured in continuous or in stationary culture with Methanobacterium strain AZ, fructose instead of being converted to 3 mol of acetate was converted to 2 mol of acetate and 1 mol each of carbon dioxide and methane, showing that interspecies hydrogen transfer occurred. In continous culture the organisms formed a close physical association in clumps; the doubling time for each organism was 6h at 33°C. Methane mainly was derived from carbon positions 3 and 4 of the sugar, but other carbons also yielded methane; this was shown to be due to carbon dioxide-acetate exchange reactions by A. woodii in a manner similar to that carried out by Clostridium thermoaceticum. Four other methanogens, Methanobacterium M.o.H. and M.o.H. G, Methanobacterium formicicum, and Methanosarcina barkeri (not acetate-adapted) also produced similar results, when co-cultured with A. woodii.

The distribution of the NADPH regenerating mannitol cycle among fungal species.

Abstract: The mannitol cycle is an important NADPH regenerating system in Alternaria alternata. The cycle is built up of the following enzymes: mannitol 1-phosphate dehydrogenase, mannitol 1-phosphatase, mannitol dehydrogenase and hexokinase. The net reaction of one cycle turn is: NADH+NADP++ATP → NAD++NADPH+ADP+Pi. The enzymes needed for an operating cycle were found in Aspergillus, Botrytis, Penicillium, Pyricularia, Trichothecium, Cladosporium and Thermomyces all genera belonging to Fungi Imperfecti. The only genus of this class lacking the cycle was Candida. No genera from the classes Basidiomycetes and Phycomycetes showed any mannitol 1-phosphate dehydrogenase or mannitol 1-phosphatase activities. The genera investigated, belonging to Ascomycetes, Gibberella, Ceratocystis and Neurospora all lacked mannitol 1-phosphate dehydrogenase. It was concluded that the mannitol cycle is an important and widespread pathway for NADH oxidation and NADP+ reduction in the organisms belonging to the class Fungi Imperfecti.

Lindane degradation by cell-free extracts of Clostridium rectum

Abstract: For lindane degradation, a cell suspension of Clostridium rectum strain S-17 demands the addition of substrates such as leucine, alanine, pyruvate, a leucine-proline mixture, and molecular hydrogen. In the presence of leucine-proline mixture, lindane decomposed in parallel with isovaleric acid formation, and both lindane degradation and isovaleric acid formation were inhibited by monoiodoacetic acid, suggesting a close relation between lindane degradation and the Stickland reaction. Lindane was degraded by cell-free extracts of C. rectum in the presence of dithiothreitol (DTT). Radiogaschromatograms of n-hexane soluble metabolites from [14C] lindane showed the presence of monochlorobenzene and γ-3,4,5,6-tetrachlorocyclohexene, Leucine, NADH, and NADPH were somewhat less active than DTT for lindane degradation in cell-free extracts. Reductive dechlorination seemed the major route of lindane degradation in cell-free extracts as well as in the intact cells of C. rectum.

Acetyl CoA, a central intermediate of autotrophic CO2 fixation in Methanobacterium thermoautotrophicum

Abstract: The pathway of autotrophic CO2 fixation in Methanobacterium thermoautotrophicum has been investigated by long term labelling of the organism with isotopic acetate and pyruvate while exponentially growing on H2 plus CO2. Maximally 2% of the cell carbon were derived from exogeneous tracer, 98% were synthesized from CO2. Since growth was obviously autotrophic the labelled compounds functioned as tracers of the cellular acetyl CoA and pyruvate pool during cell carbon synthesis from CO2.

Regulation of chitin synthesis during germ-tube formation in Candida albicans

Abstract: The synthesis of chitin during germ-tube formation in Candida albicans may be regulated by the first and last steps in the chitin pathway: namely l-glutamine-d-fructose-6-phosphate aminotransferase and chitin synthase. Induction of germ-tube formation with either glucose and glutamine or serum was accompanied by a 4-fold increase in the specific activity of the aminotransferase. Chitin synthase in C. albicans is synthesized as a proenzyme. N-acetyl glucosamine increased the enzymic activity of the activated enzyme 3-fold and the enzyme exhibited positive co-operativity with the substrate, UDP-N-acetylglucosamine. Although chitin synthase was inhibited by polyoxin D (Ki =1.2μM) this antibiotic did not affect germination. During germ-tube formation the total chitin synthase activity increased 1.4-fold and the expressed activity (in vivo activated proenzyme) increased 5-fold. These results could account for the reported 5-fold increase in chitin content observed during the yeast to mycelial transformation.

The effects of light quality on the growth of phototrophic bacteria in lakes

Abstract: The effects of light quality (color) on the 14CO2 fixation rates of natural population of photosynthetic sulfur bacteria were tested. The phototrophic bacteria were collected from the sulfide containing waters of 3 stratified lakes. The populations sampled survive in environments where light intensities are very low. Not only are the light intensities low but, due to the “light filtering” characteristics of the lake water, the light is of specific color. It was determined that the spectral properties of the three lakes differed, hence the quality of light reaching the phototrophic bacteria in each lake differed. It was also observed that only green sulfur bacteria were present in the study lake which transmits mainly red light and both purple and green sulfur bacteria were present in the two study lakes which transmit predominantly green light. Enrichment cultures were set up with phototrophic bacteria from two of the lakes serving as the inocula. Enrichment culture studies and photosynthetic responses of the natural populations indicate that light quality is a major factor in determining the composition of phototrophic bacterial population in some lakes.

Mixotrophic growth of Thiobacillus A2 on acetate and thiosulfate as growth limiting substrates in the chemostat

Abstract: During heterotrophic growth on acetate, in batch culture, the autotrophic growth potential of Thiobacillus A2, i.e. the capacity to oxidize thiosulfate and to fix carbon dioxide via the Calvin cycle, was completely repressed. The presence of thiosulfate in a batch culture with acetate as the organic substrate partly released the repression of the thiosulfate oxidizing system. Cultivation of the organism in continuous culture at a dilution rate of 0.05 h-1 with different concentration ratios of thiosulfate and acetate in the reservoir medium led to mixotrophic growth under dual substrate limitation. Growth on the different mixtures of acetate and thiosulfate yielded upto 30% more cell dry weight than predicted from the growth yields on comparable amounts of these substrates separately. The extent to which the carbon dioxide fixation capacity and the maximum thiosulfate and acetate oxidation capacity are repressed appeared to be a function of the thiosulfate to acetate concentration ratio in the reservoir medium. The results of 14C-acetate assimilation experiments and of gas-analysis demonstrated that the extent to which acetate was assimilated depended also on the substrate ratio in the inflowing medium. Under the different growth conditions surprisingly little variation was found in some tri-carboxylic acid cycle enzyme activities. Cultivation of T. A2 at different growth rates with a fixed mixture of thiosulfate (18 mM) and acetate (11 mM) in the medium, showed that dual substrate limitation occured at dilution rates ranging from 0.03–0.20 h-1.

Oxygen utilization by Lactobacillus plantarum

Abstract: Lactobacillus plantarum (ATCC 8014) cells, grown aerobically on glucose medium, consumed molecular oxygen when incubated with either glucose, d/l-lactate or pyruvate as substrate. Cell extracts catalyzed the oxidation of NADH, d/l-lactate or pyruvate with O2. Per mol O2 2mol of NADH were consumed indicating that O2 was reduced to H2O; reduction proceeded via H2O2 involving a NADH oxidase and a NADH peroxidase. Catalase activity was absent. Pyruvate oxidation with O2 led to the formation of H2O2, lactate oxidation to the formation of H2O. Thus in L. plantarum different mechanisms are available by which molecular oxygen can be used as electron acceptor for oxidation reactions.

ATP-linked citrate lyase activity in the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum

Abstract: Cell-free extracts of the green sulfur bacterium Chlorobium limicola forma thiosulfatophilum strains 1C and L have been shown to cleave citrate with the formation of oxaloacetate and acetyl-CoA. This capacity was found in autotrophically grown cells as well as in the cells grown on media with acetate or L-glutamate. Citrate lyase activity in cell-free extracts is only measurable in the presence of citrate, adenosine-5′-triphosphate, coenzyme A and Mg2+ or Mn2+. It is concluded on the basis of the obtained data that C. limicola f. thiosulfatophilum contains adenosine-5′-triphosphate-linked citrate lyase (E.C.4.1.3.8). In contrast to green bacteria in the purple bacteria Ectothiorhodospira shaposhnikovii, Rhodospirillum rubrum and Thiocapsa roseopersicina citrate lyase activity was not found.