Showing papers in "Journal of General and Applied Microbiology in 1969"

Occurrence of plasmalogens in anaerobic bacteria

Abstract: YOSHIYUKI KAMIO, SHIRO KANEGASAKI AND HAJIME TAKAHASHI Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University, Sendai, Japan (Received April 14, 1969) Plasmalogens, aldehyde-containing phospholipids, were found to occur widely in all four fractions of ruminal microbes obtained after differential centrifugation of sheep rumen contents. Plasmalogens were also detected in ruminal and soil anaerobic bacteria which were obtained by enrichment cultures with various energy sources. They occurred widely in all strictly anaerobic bacteria and Propionibacterium tested. The strains included Desul fovibrio sp., Selenomonas ruminantium, Bacteroides ruminicola, Veillonel-la gazogenes, Peptostreptococcus elsdenii, Propionibacterium freudenreichii, Pro-pionibacterium shermanii, Clostridium saccharoperbutylacetonicum, Clostridium acetobutylicum, Clostridium perfrigens, Clostridium kaneboi, and Clostridium kainantoi. Although the ubiquity of plasmalogens in these anaerobic bac-teria was confirmed, the amount of plasmalogens per unit weight of cells varied considerably depending on bacterial species. Amount of plasmalogens in Selenomonas and Propionibacterium also varied by cultural conditions. However, plasmalogens were not detected in aerobic bacteria and in facultative anaerobic bacteria even when they were cultured under strictly anaerobic conditions.

PARTICIPATION OF A REGULATOR GENE IN THE α-AMYLASE PRODUCTION OF BACILLUS SUBTILIS

Abstract: Bacillus natto IAM 1212 produces α-amylase by about 5 times as much as Bacillus subtilis Marburg strain, α-amylase of which is more thermostable and moves more rapidly to anode at pH 8.3 than that of B. natto. When this high producing activity of α-amylase (Amysup character) was transferred to a Marburg strain by the DNA from B. natto, about one-tenth of the transformants with Amysup character produced recipient type, i.e. Marburg type α-amylase and the other natto type α-amylase. In other words, the former acquired only high producing activity of α-amylase. This result suggests that a gene which regulates α-amylase synthesis participates in the α-amylase-producing system of B. subtilis and this regulator gene (amyR) is closely linked to the α-amylase structure gene. Reciprocal crossing also confirmed this assumption. The amyR gene is linked to the aro116 gene which has been reported to be the linked marker of α-amylase structure gene by S. Yuki.

The influence of nitrogen and carbon sources on the production of glucoamylase by aspergilli

Abstract: Glucoamylase produced by Aspergillus niger was observed to increase in culture media at two specific intervals, namely, after three days and after six days of incubation. The lowest levels of enzyme production occurred when the nitrogen source was asparagine, trypticase or urea. Intermediate quantities of enzyme were formed when the fungi were grown on a nitrogen source of yeast extract. Corn steeping liquor and nutrient broth (2.5%) gave highest enzyme activity. The inorganic nitrogen source providing the best glucoamylase yield was ammonium-N. Nitrate-N or a mixture of ammonium-N and nitrate-N was less effective. When inorganic nitrogen was employed, the two periods of glucoamylase production were not as readily demonstrated. With ammonium-N, all the glucoamylase is produced during the fourth or fifth day of incubation, thus showing only one period of enzyme synthesis.The quantity of glucoamylase produced depended upon the monosaccharide added to the growth medium. Glucose yielded the highest level of enzyme regardless of the nitrogen sources. Mannose, usually, was slightly more effective than sorbitol and produced about half the quantity of glucoamylase obtained from glucose. Lowest levels of enzyme occurred when fructose or xylose served as carbon sources. Growth on 1.0 or 2.0% carbohydrate, rather than 5.0%, was found to result in higher levels of glucoamylase. Although A. niger NRRL 330 was the principle organism tested, the characteristics of glucoamylase synthesis were not specific for this strain since similar results were obtained with A. niger NRRL 3122 and A. awamori NRRL 3112.

Taxonomic studies on coryneform bacteria:i. division of bacterial cells

Abstract: Cell division of the so-called eubacteria and coryneform bacteria was studied by the time lapse microscopy and microcinematography. The rodshaped bacteria were divided into the three types of simple type, snapping type, and bending type on the basis of the mode of cell division. From the results obtained the mode of bacterial cell division is considered to indicate a taxonomic significance, and eubacteria capable of growing by simple type division are clearly distinguished from coryneform bacteria which multiply by snapping or bending type.

Cold shock of bacteria

Abstract: Escherichia coli, Pseudomonas fluorescens, and Bacillus subtilis cells, harvested at the logarithmic phase of growth, were susceptible to cold shock as judged by their loss in viabilities. Viabilities of cold shocked cells increased rapidly upon incubation at 30° with suitable additives. Among the effective agents, magnesium ion was the most important, because no appreciable recovery in viability was observed in the absence of magnesium. When the cold shocked E. coli cells were incubated at 30° with 5×10-3M of magnesium acetate for 10 to 20min, the viability of shocked cells became equal to that of unshocked cells. The addition of 2, 4-dinitrophenol to such systems inhibited the magnesium-mediated recovery almost completely. When NAD or ATP plus nicotinamide was further added to the reaction mixture, the inhibition by 2, 4-dinitrophenol was released. This result suggests that NAD, possibly as an energy source, is involved in the recovery process.An increase in permeability of cold shocked E. coli cells was demonstrated with the aid of 8-anilino-1-naphthalenesulfonate. The stimulatory effect of NAD or ATP added to the reaction mixture on the recovery from the cold shock could be explained by this permeability increase.The presence of two critical temperature zones in the cold shock was confirmed with both B. subtilis and Ps. Fluorescens. When the initial temperature of the cell suspension before the cold shock was lowered by 5°, both temperature zones moved to lower temperatures approximately by 3-5°.

A new sulfate-reducing bacterium isolated from antarctica

Abstract: Determinative studies were carried out with an obligatory anaerobic spore-forming sulfate-reducing bacterium, strain No. 64, isolated from the SKARVS NES district in the East Antarctica. This sulfate-reducing bacterium belongs to the genus Desulfotomaculum differing from all of the known species of the genus in respect to glucose fermentation, gelatin liquefaction, optimum temperature for growth, and nutritional requirements. The strain was determined to be a new species of the genus Desulfotomaculum, D. antarcticum, IIZUKA et OKAZAKI nov. sp.

Thin-layer chromatography of some c21, c19 and c18 steroids

Abstract: The resolution of 30 different C21, C19 and C18 steroids by various solvent systems on silica gel chromatoplates was reported. Effective resolution of Δ4-pregnene, 5α, axial-equatorial hydroxyl epimers and positional isomers was successfully achieved. The differentiation between these compounds by using four different colour reagents has been also reported.

Factors influencing the production and further metabolism of indole-3-acetic acid by pseudomonas savastanoi

Abstract: Certain factors which affect production and accumulation of indole-3- acetic acid (IAA) and activity of enzymes associated with its synthesis from L-tryptophan were investigated in Pseudomonas savastanoi. Addition of L- tryptophan to the culture medium enhances the accumulation of IAA by the bacterium. However, both tryptophan oxidative decarboxylase and indoleacetamide hydrolase, which are involved in the conversion of L-tryptophan to IAA, are produced by the bacterium without addition of L-tryptophan to the medium. In a medium containing L-tryptophan, yields of tryptophan oxidative decarboxylase per liter of culture suspension were increased as much as 2-fold.IAA and indole-3-acetamide (IAM) strongly inhibit tryptophan oxidative decarboxylase. IAA up to 6×10-3M did not inhibit indoleacetamide hydrolase. IAA also inhibits uptake of L-tryptophan by whole cells.IAA is converted to its lysine conjugate by the bacterium. During logarithmic phase of growth in a nutrient medium, the specific activities of tryptophan oxidative decarboxylase and indoleacetamide hydrolase were 2-fold greater than that for the system converting IAA to its lysine conjugate. During the stationary growth phase, specific activity of all three enzymes decreased but that for the first two enzymes did so more rapidly than the specific activity of the lysine conjugate synthetase. Accumulation of IAA in culture filtrates is controlled to a large extent by the relative activities of the systems synthesizing and utilizing IAA.

Studies on the aggregation of yeast caused by lactobacilli

Abstract: Aggregation of yeast by lactobacilli was studied by a method of boundary moving electrophoresis.The isoelectric point of sake yeast and lactobacillus B-83 was at pH between 3.5 and 4, while that of wine, alcohol, brewer's, and baker's yeasts and lactobacillus B-74 was below pH 2.The aggregation was observed in the combination of the microorganisms which had different isoelectric points and in the pH range in which the signs of the cell surface charge of yeast and lactobacilli are contrary.From these results it was supposed that aggregation of yeast by lactobacilli would be due to the electrostatic force between yeast and bacterial cell surfaces.

Genetic and biochemical studies on bacterial formation of l-glutamate

Abstract: 1. One acetate kinase-negative, 53 phosphate acetyltransferase-negative, and seven isocitrate lyase-negative strains were found among 73 mutants of Brevibacterium flavum, which grew on glucose but not on acetate. The result indicates that these enzymes are essential for the utilization of acetate in B. flavum, and also that the participation of acetyl-CoA synthetase is absent in this strain.2. The genetic defect of any one of these three enzymes had no effect on the production of L-glutamate from glucose.3. Many mutants possessing a variety of isocitrate lyase activities were obtained by deriving revertants grown on acetate from four isocitrate lyase-negative strains. By an experiment using such revertants, it was concluded that isocitrate lyase had an important effect on the production of L-glutamate from acetate, which was shown as a positive correlation between the activity of this enzyme and L-glutamate productivity in the cells grown in an acetate medium. It was also shown that the change of this enzyme activity had no effect on the production of L-glutamate from glucose.4. Mutants possessing a variety of phosphate acetyltransferase activities were obtained by deriving revertants grown on acetate from four strains lacking this enzyme. In this case, too, a positive correlation was observed between the activity of this enzyme and L-glutamate productivity in the cells grown in an acetate medium.5. In connection with the present study, it was pointed out that genetic elevation of the levels of key enzymes was important for the improvement of microbial strains for industrial purposes.

Comparative study on pediococcus halophilus, p. soyae, p. homari, p. urinae-equi and related species

Abstract: A comparative study was made on three salt-tolerant, dextro-rotatory lactic acid-forming pediococci, Pediococcus halophilus, P soyae, and P homari, and related species, P urinae-equi and Aerococcus viridans, with reference to P pentosaceus and P acidilactici The former three species were sufficiently similar in their morphological, physiological, and nutritional characteristics, and DNA Tm value to place them into a single species P urinaeequi is almost similar to them, especially nutritionally, but differed from them only on the point of salt sensitivity It was proposed that their boundary be the growth on 12% salted medium A viridans has characters much similar to P urinae-equi to be classified within the species, although its preference for aerobic conditions and other non-fermentative characters suggest its relation with Staphylococcus-Micrococcus group The usefulness of nutritional requirement for the classification of this genus was discussed

Arthrobacter luteus nov. sp. isolated from brewery sewage

Abstract: A group of bacteria isolated from brewery sewage was studied taxonomically They were gram-positive, facultatively anaerobic, pleomorphic, branching, non-motile, non-sporulating, non-acid-fast, and catalase-positive rods (06-10×08-100μ) They formed cystites and showed bending-type cell division They produced a yellow pigment and reduced nitrate, hydrolyzed starch, and liquefied gelatin They produced acids from various carbohydrates These characteristics were compared with those of 18 strains of related microorganisms The isolates seemed to belong to the genus Arthrobacter, but no corresponding species was found in the taxa appearing in Bergey's Manual (7th Ed) The name Arthrobacter luteus was, therefore, newly proposed for these isolatesWhile the new species was in accord with those of the genus Arthrobacter in basic characteristics, it also had similarities to some species of the genera Nocardia, Cellulomonas, Microbacterium, and CorynebacteriumThese observations suggested that the species occupied an intermediate position between the families Corynebacteriaceae and Actinomycetaceae

Microbiological transformations of progesterone

Abstract: Transformation of progesterone by various microorganisms such as actinomycetes, yeast and mould fungi of different 53 strains had been studied. 19 different strains was able to oxygenate progesterone at C11. The transformation products, other than 11α-hydroxyprogesterone, which were formed by these active strains had been identified. These studies proved that the active organisms were capable of introduction of oxygen function at different positions other than position 11 of progesterone e.g. positions 17 and 21. Side chain degradation reaction with and without lactone formation of ring D of progesterone had been also found to be performed by some of these organisms.

Dna base composition of the genus hansenula

Abstract: The DNA base ratio (GC content) of 46 cultures of Hansenula and allied yeasts which represented 26 species or varieties was studied The GC contentof Hansenula DNA ranged from 285 to 463% Intrageneric variation reached 156-178% in Hansenula sensu WICKERHAM, 144-158% in Hansenula sensu LODDER and KREGER-VAN RIJ, 134-148% in Hansenula emended by NOVAK and ZSOLT, and 110-127% in Hansenula group (genus) of TSUCHIYA et al The frequency curve of the DNA base ratio demonstrated the presence of two large groups in the genus Hansenula The first group had relatively high values of GC content with maximal frequency in a range of 40-42% The second had relatively low values of GC content with the maximal frequency in a range of 32-34% Species arranged in each line of the phylogenetic scheme of Hansenula proposed by WICKERHAM exhibited consistent DNA base ratios individually though exceptions were found, and so did the serological groups within Hansenula group (genus) of TSUCHIYA et al A taxonomic and phylogenetic evaluation of DNA base ratio was discussed in relation to the delimitation of the genus Hansenula by several workers and the existence of several groups having similar DNA base ratio within the genus

Studies on (-)-citramalic acid formation by respiration-deficient yeast mutants:v. purification and some properties of citramalate condensing enzyme

Abstract: Citramalate condensing enzyme has been purified 95-fold by ammonium sulfate fractionation, and by Sepharose 4B and DEAE-cellulose column chromatography from RD petite mutant strain of Saccharomyces carlsbergensis. On the basis of the fact that (-)-citramalate fraction had all of the radioactivity while (+)-citramalate fraction contained no radioactivity when enzymically formed radioactive citramalic acid with authentic carrier DL-citramalic acid was subjected to optical resolution, it was made clear that this enzyme catalyzed the formation of (-)-citramalate from pyruvate and acetyl-CoA. The optimal pH of the enzyme was 7.4 and the Km value for pyruvate was 2.3×10-3M. The purified enzyme preparation still has an activity toward α-ketobutyrate and α-ketoisovalerate. The Km value for α-ketoisovalerate was 2.6×10-5M. L-Leucine inhibits to the same extent the respective condensation reactions between these three α-keto acids and acetyl-CoA. α-Ketoisovalerate is an effective inhibitor of citramalate condensing reaction. The enzyme was strongly inhibited by p-chloromercuribenzoate, Cu2+, Zn2+, Hg2+, Pb2+, and Cd2+. Citramalate condensing enzyme appears to be identical with α-isopropylmalate synthetase in the leucine biosynthetic pathway. The role of this enzyme in RD petite mutant strains of Saccharomyces carlsbergensis was discussed in relation to citramalate formation.

Fungi of delhi

Abstract: Two rare fungi isolated from the soils of Delhi have been described. Aspergillus penicilliformis KAMYSCHKO seems to be a link between the enera Aspergillus and Penicillium. Preussia aurantiaca RAI and TEWARI has been transferred to Pycnidiophora aurantiaca (RAI & TEWARI) MUKERJI and RAO comb. nov. A key to the genus Pycnidiophora Clum is included.

Utilization of gas oil in the production of single cell protein

Abstract: 1. For the production of single-cell protein, isolation of yeast strains from oil-soaked soils and water was carried out. Out of several yeasts isolated, nine were identified and found to belong to the genus of Candida, Trichosporon, Pichia, and Saccharomyces.2. Strains of Candida and Trichosporon grow well in gas oil with high specific growth rates.3. Crude protein content of the dried cells vary between 48 and 62%.

Properties of toxins of vertigillium dahliae, the causative agent of cotton wilt disease

Abstract: 1. Determination of the toxicity of the culture liquid of Verticillium dahliae showed that it contains an active toxin which causes wilting of cotton plant shoots. Thus the fungus produces a toxin which it excretes into the external medium.2. By testing a large number of strains of microorganisms from various taxonomic groups, a test microbe was found which is sensitive to the toxin of Verticillium dahliae.3. Such a sensitive test-microbe made it possible to use chromatography with a bioassay method to select the best solvent for extraction of the toxin.4. The toxin was obtained in concentrated form by extraction with amyl acetate.5. A study of the physico-chemical properties of the toxin showed it to be acidic in nature, and that it has two sharp absorption maxima in the UV region at 245 and 275mμ, and is stable at high temperatures and over a wide range of pH values.6. The toxin of Verticillium dahliae is composed of two fractions, differing in physico-chemical and biological properties.7. The toxin of Verticillium dahliae does not appear to be highly specific, and, besides its marked inhibitory action on cotton, causes wilting of leguminous plants, during which time cereals are not sensitive to the toxin in the concentrations tested.8. The rate of wilting under the influence of the toxin depends on the age and variety of cotton plant. Adult plants wilt more quickly than young plants. The cotton plant variety 108-F (resistant to wilting) is less sensitive to the toxin than variety 2034, which is non-resistant to wilting. The toxin causes browning of the vascular bundles and the surrounding woody parts of the stalk, and the leaves loose their turgidity.9. The tetraene antibiotics, 18-45 and 18-80, produced by actinomycetes, have an effect on the toxin produced by Verticillium dahliae. They interact with the fungal toxin and neutralise its action.10. Treatment of seeds with antibiotics or cultivation in soils which have been enriched with the antibiotic-producing actinomyces produces an immunity in the cotton plants which is passed on to the following generations.

Histochemical studies on the development of carpophore of coprinus kimurae

Abstract: A process of development of Coprinus kimurae HONGO et AOKI cultured in this laboratory was morphologically and histochemically observed. The process was tentatively divided into seven stages according to morphogenesis. Activities of cytochrome oxidase, succinic dehydrogenase, and acid and alkaline phosphatases were mainly found at active growth zones of this mushroom such as top part of stalk and margin of cap. Both kinds of phosphatases were not detected in gills at stages 3 and 4 but were detected in gills at stage 5 when cap grew remarkably and spores formed.

Studies on sporulation and production of mycobacillin, an antifungal polypeptide, by bacillus subtilis

Abstract: Both mycobacillin production and sporulation are initiated in the postlog phase of growth of B. subtilis B3, reaching their maximum by about the same time. Mycobacillin is released during enndotropic sporulation of vegetative cells though not during germination of spores in complex growth medium. Antisporogenic chemicals like glucose (in excess), diethylmalonate, acriflavin, fluoroacetic acid, sodium bisulphite, β-phenethyl alcohol, α-picolinic acid and m-tyrosine inhibit mycobacillin production. By acriflavin and actinomycin D treatment, two types of mutants, oligosporous and asporogenous, were obtained. Mycobacillin production is affected adversely in oligosporous mutants whereas asporogenous mutants do not produce the antibiotic at all.

Molecular mechanism of gram staining

Abstract: The passage of small molecular substances through the walls of Staphylococcus aureus is reduced in presence of 95% ethanol, as well as solutions of aniline, dimethyl aniline and nitrobenzene in 95% ethanol. But these three solvents, unlike 95% ethanol alone, can extract dye from Gram-stained Staphylococcus aureus almost completely, indicating the inadequacy of the permeability theory of Gram staining.It has been observed that, if iodine from Gram-stained Staphylococcus aureus is removed by thiosulphate, the retained dye can be extracted by 95% ethanol. However, if after thiosulphate treatment Staphylococcus aureus with the retained dye is treated with graded amounts of iodine, extractibility of the dye by 95% ethanol is progressively reduced. This supports the model proposed for the mechanism of Gram staining which involves retention of crystal violet by the cell component presumably by electrostatic bonds and formation of chargetransfer complex of crystal violet and iodine through π-electrons. Stability of this cell component-dye-iodine complex towards 95% ethanol determines the Gram character of the cell.

Protonic conduction of artificial lipid membranes

Abstract: Electrical conductance of phospholipid bilayer membranes formed in various electrolyte solutions was studied with special reference to the kind of related charge carriers. The conductance was extremely low (10-5-10-7mho-cm-2), depending considerably on the lipid composition but little on both the concentration and kind of the electrolytes used, and it showed a complex dependence on the pH of the media on both sides of the membrane. Because of these experimental results, it seems preferable to assume that the charge carriers through the membrane are not the ions supplied from the electrolyte added to the solution, but protons and hydroxyl ions whose mobilities depend upon the sign and magnitude of the net charge on the membrane. Potential differences caused by diffusion of protons or hydroxyl ions across the membrane were also determined to prove the above assumption.

Oxidative decarboxylation of 6-phosphogluconate in leuconostoc mesenteroides b07:ii. purification and properties of a new enzyme catalyzing decarboxylation of 2-keto-6-phosphogluconate

Abstract: Although 6-phosphogluconate dehydrogenase (decarboxylating) (EC 1.1.1.44) has not been detected in Leuconostoc mesenteroides B07, the NAD-linked dehydrogenase catalyzing only oxidation of 6-phosphogluconate was purified from this organism.Basal properties of the dehydrogenase was studied and the dehydrogenation product was isolated and identified as 2-keto-6-phosphogluconate.