Zeitschrift für allgemeine Mikrobiologie

Published by John Wiley and Sons
Online ISSN: 0044-2208
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Article
In sporenbildenden aeroben Bakterien ist die Sekretion verschiedener Exoenzyme an das Erreichen eines bestimmten physiologischen Zustandes, der während des Sporenstadiums 0-1 erreicht wird, gebunden. Neben Ribonuclease, alpha-Amylase, neutraler and alkalischer Protease konnte in der Kulturflüssigkeit von Bacillus subtilis ein weiteres Enzym, eine ß-1,3-1,4-Glucanase nachgewiesen werden. Das Ferment katalysiert die hydrolytische Splaltung der inneren ß-1,3- und ß-1,4-Bindungen von ß-Glucanen, die aus Cerealien oder aus Cetraria islandica, dem Isländischen Moos, isoliert werden können. Dabei entstehen als hauptsächliche Hydrolyseprodukte 3-O-ß-D-Cellobiosyl-D-Glucose und 3-O-ß-D-Cellotriosyl-D-Glucose. In der vorliegenden Arbeit wird die Bildungskinetik der ß-Glucanase im Verlaufe des Wachstums im Zusammenhang mit verschiedenen Indikatoren der Sporenreifung verfolgt. Die ß-Glucanasebildung beginnt am Ende des exponentiellen Wachstums. Sie wird in der beginnenden stationären Phase mit konstanter Geschwindigkeit fortgesetzt und kommt erst nach 18 h Kulturdauer unter den gewählten Wachstumsbedingungen zum Stillstand. Ein starker Anstieg der Anzahl hitzeresistenter sporen wurde am Ende der Glucanaseproduktion nach 16 h Kulturdauer beobachtet. Die Sporenreifung war erst nach 25 h unter unseren Bedingungen abgeschlossen. Die hier mitgeteilten Resultate weisen darauf hin, dass die ß-Glucanase im Sporenstadium 0-1 gebildet wird.
 
Article
In the agar diffusion test 24 triazines were investigated with regard to their action on the mulplication of DNA phages (lambda and LPP-1) and RNA phages (M12 and Qbeta). In several cases the amount of plaques was diminished or increased depending on the kind of triazine and virus. The investigations demonstrate the triazines to be able to interfere with the formation of plaques by virulent and temperate viruses of procaryotes.
 
Article
(20S)‐20‐Carboxy‐1,4‐pregnadien‐3‐one was hydroxylated by Cochliobolus lunatus in positions 11β and 12β. The two novel compounds were characterized by means of UV‐, IR‐ and NMR‐spectroscopy. The possibility to use microbial degradation products of the 20‐carboxy‐pregnane type for the partial synthesis of glucocorticosteroids is discussed.
 
Article
Fructose-1,6-bisphosphate aldolase (Fru-P2A) from a psychrophilic marine bacterium was found to be Class II aldolase based on activation by K+, activation by divalent cations, inactivation by EDTA, low molecular weight, and similar values for Km, Vmax, and Arrhenius activation energy. This enzyme was not markedly different in amino acid composition from the enzymes from mesophilic and thermophilic organisms, yet it has unusual thermal properties.
 
Article
A Bacillus amyloliquefaciens strain high-producing for beta-1.3-1.4-glucanase has gradually lost the ability to produce this enzyme during long-time continuous cultivation, independent of the culture conditions. Mutant strains isolated after long-term cultivation exhibited changed behaviour concerning extracellular enzyme formation and sporulation. By agarose gel electrophoresis of alkaline DNA extracts isolated form original and mutant strains we demonstrate that the observed pleiotropic phenomena are not caused by the loss of a complete plasmid present in the original strain. From extracts of both the original and mutant strains plasmid DNAs with approximately the same molecular weight of about 35 Mdal were isolated.
 
Article
Cell-free extracts from Candida rugosa JF 101, disrupted with a cell homogenizer, oxidized 1-decene to decyl alcohol, decyl aldehyde and decanoic acid under aerobic and anaerobic conditions. A linear relation' between enzyme concentration and the amount of the decyl alcohol formed was observed. The anaerobic concentration of decyl alcohol was approximately 2 to 3 times greater than under aerobic conditions. By use of mass spectrometry it was shown that the oxygen of water is incorporated into the 1-decene during hydroxylation. From these results, the most probable mechanism would be hydration of 1-decene to decyl alcohol by an enzyme analogue to fumarase. A tentative mechanism for n-decane hydroxylation in C-rugosa JF 101 system is proposed.
 
Article
In laboratory-scale experiments with growing cells of the obligate methane-oxidizing strain M 102, an overall molar gas turnover ratio of the order given below could be postulated: 1 CH4+1--1.2 O2=0.3 CO2+water. Expectations that the optimal gas mixture of methane and oxygen should lie within the range of this stoichiometric consumption ratio have been verified in fermenter 5 1 batch culture experiments. The optimal range of methane-oxygen mixture, found under the experimental conditions described, is based on the estimated growth parameters as generation and doubling times, yield coefficients related to methane and oxygen, and the efficiency of methane metabolism as indicated in the absolute amounts of CH4, O2, and CO2 turned over. The mentioned stoichiometric relation of 1 CH4:1--1.202 did not change with varying the composition, i.e. the partial pressures of CH4 and O2 introduced as a mixture to the cells. The efficiency of methane oxidation was obviously influenced and decreased markedly when deviating from the broad optimal range of CH4/O2 mixtures. With non-growing cells, on the other hand, the stoichiometric relation showed a considerable shift (1:1.4--1.8 CH4:O2) with a clear tendency towards more O2 consumption. The oxidation potential of growing cells, seems then to have a linear interdependence to the substrate concentrations, i.e. partial pressures.
 
Article
Proteus mirabilis strain 1095/67 has been cultivated in surface culture on different minimal media, which were composed either of single amino acids or combinations of several amino acids without or with stepwise addition of other C-sources up to the complete medium. We found a direct dependence of the formation of the various antigens (moving to the cathode, to the anode, or not moving) on the composition of the medium. Certain amino acids (Ala, Glu, Asp. Ser) elicit only parts of the antigenic spectrum formed in complete medium, but enrich special antigens. With only 2 amino acids (Ala and Glu) and the vitamins thiamine, pyridoxal, and niacine in the anorganic basic medium the complete antigenic spectrum can be formed. We have shown that the composition of the cell wall varies with the composition of the culture medium. In shaked cultures we found serine to be essential for the production of the antigen moving to the cathode and the thermolabile antigene moving to the anode (ATA), niacine to be of fundamental importance in the biosynthesis of all antigens. Already under very limited supply of nutrients ATA, common to all gram-negative bacteria, is formed and, therefore, it seems to be a very basic cell wall component.
 
Article
In the presence of malate or citrate sporangiospores of C. elegans were able to hydroxylate cortexolone with a rate twofold exceeding that of the control, water suspended spores. Analysis of the intracellular nicotinamide coenzyme pools revealed an increased NADPH: (NADP+ + NADPH) ratio, indicating more effective NADPH-generating systems in malate- or citrate-stimulating spores. Swollen spores remaining in the pregermination state, retained higher cortexolone-hydroxylating activity in the absence of malate and citrate. In these spores degradation of endogenous alanine and glutamic acid was observed. Possible NADPH-generating systems in C. elegans sporangiospores were discussed.
 
Article
Two newly isolated aniline-degrading bacterial strains were characterized with regard to their enzyme systems responsible for aniline catabolism. One of them identified as a Rhodococcus sp. metabolized aniline exclusively via the beta-ketoadipate pathway by means of inducible enzymes. The aniline-degrading enzyme system of the second isolate, presumably a pseudomonad, was shown to consist of an inducible aniline-converting enzyme and constitutive meta-pathway enzymes. Both isolates failed to metabolize monochlorinated anilines in the absence of additional carbon sources. To explain this the ring-cleaving enzymes of both isolates were examined for their substrate specificities. Furthermore, the effect of 4-chlorocatechol on the enzymes catalyzing aniline conversion and catechol oxygenation was investigated.
 
Article
The capsule material of Staphylococcus aureus strain 1193/74 could be separated by precipitation with trichloroacetic acid and ethanol as well as by chromatography on DEAE-cellulose into 13 fractions. All fractions contained saccharides and uronic acids as well as amino acids and appeared in their qualitative composition rather similar. However, in quantitative composition and in chromatographic behaviour a rather high degree of heterogeneity could be observed. No clear cut separation of protein and polysaccharide material could be achieved in any fraction. It is supposed, therefore, that the capsule material does not represent merely an acidic polysaccharide, but contains certain amounts of amino acids or peptides varying in a rather wide range.
 
Article
Hydroxylation of progesterone at C-6, C-11, C-17, C-21 as well as formation of 11α-hydroxyallopregnane-3,20-dione was successfully performed with cell homogenate of a local strain of Rhizopus nigricans REF 129. Progesterone was only transformed to 17α- and 21-hydroxy derivatives with the supernatant solution of the cell homogenate. 5α-Hydrogenase, β-hydroxylase, and 11α-hydroxylase appeared to be located in the cell-sediment fraction. Successful isolation of a cell preparation containing 11α-hydroxylase system was obtained by treating the cell-sediment fraction with citrate buffer, then by precipitation with ammonium sulphate. The formation of 6β,11α-dihydroxyprogesterone was hindered by the addition of ascorbic acid to the reaction mixture.
 
Article
From the partial hydrolysates of the lipopolysaccharides from strains 2 (Sh. flexneri 1b) and 1290/63 (Sh. flexneri var. Y) we isolated in both cases a Rha-Rha-disaccharide. After trimethylsilylation, gas liquid chromatographic purification and mass spectrometry we could show that the rhamnose molecules of both disaccharides were linked to each other by a 1,3-glycosidic bond. As we could estimate the other linkages in the O-specific side chains of these lipopolysaccharides in recent studies, we are now able to present the following structures for the repeating units: In addition, we found in the lipopolysaccharide of strain 1290/63 rather big quantities of a poly-α-(1,4)-glucan.
 
Article
With Candida spec. H and the mutant H13 during nitrogen starvation we studied the changes occurring in the macromolecular cell composition. In the absence of nitrogen, but in the presence of n-alkanes as carbon source, both the nucleic acid and protein levels of the dried yeasts rapidly decreased. At the same time carbohydrates and lipids were accumulated. Candida spec. H predominantly increased carbohydrate content, whereas the mutant accumulated significantly higher amounts of lipids. The main components of the cellular carbohydrates and the fatty acid content were analyzed, and the results were discussed.
 
Article
There is a rapid uptake of 14C-glucose and unlabelled glucose during the period of pigmentation of the mycelium of Botryodiplodia theobromae Pat. Incorporation of 14C-glucose into structural components of the mycelium started within 24 hours of incubation and progressively increased until the 14th day of growth. The mycelium contains glucose, isomaltose, and sucrose. There is a decrease in these carbohydrates during autolysis. Autolysis in B. theobromae occurs in an acid medium of pH 5.2.
 
Article
During application of D-glucose-u-14C paromomycine II is higher labelled and shows a different dependence on the application time than paromomycine I, which is isomer at the paromose part. For the two paromose isomeres different rates of synthesis are supposed that change nonproportionally to each other. The distribution of radioactivity in paromomycine I shows that there is no fragmentation of the glucose chain during the biosynthesis of glucosamine, ribose, and paromose I. As to the 2-deoxystreptamine the result has not been ascertained.
 
Article
Paromomycin was isolated from culture filtrates of Streptomyces albus var. metamycinus nov. var. after feeding the growing cultures with D-glucose-u-14C. From the different incorporation rates conclusions concerning different features of the paromomycin biosynthesis (utilization of the carbon source, proportional and disproportional changes of the rates of synthesis) could be drawn. Uptake and metabolism of glucose are discussed.
 
Article
Distribution of radioactivity in paromomycin ascertained after application of 14C-D-glucose, 14C-D-glucosamine, 14C-2-deoxystreptamine, respectively, 14C-D-ribose is taken as basis for a biosynthesis scheme: While ribose bound in the antibiotic originates from glucose by oxidation and following decarboxylation, glucosamine is formed via fructose-6-phosphate. Paromose I arises from glucosamine, but not the cyclohexan derivative 2-deoxystreptamine, whose biosynthesis pathway is directly branching off glucose.
 
Article
Mannose-6-p is an activator of the 14C-mannose incorporation from GDP-14C-mannose in the mono- and oligosaccharides and in the mannopolymers of the cell wall proteophosphomannan produced by the food protein yeast Candida spec. H. Moreover, mannose-6-p is a precursor of proteophosphomannan: 14C-Mannose-6-p has been incorporated in absence of GTP. Corresponding behaviour shows glucose-6-p by synthesis of beta-glucan and glycogen. Mutants of Candida spec. H with different efficiency in the biosynthesis of mannan, beta-glucan and glycogen incorporate hexose-6-p in a different extent.
 
Article
Biosynthesis of [7-3H]16alpha-hydroxy-dehydroepiandrosterone in high specific activity has been studied. [7-3H] dehydroepiandrosterone (13.9 C/mM) in trace quantity was oxidized by Streptomyces roseochromogenes (NRRLB-1233) for 5 min at 27 degrees C. The radioactive products were chromatographically separated, identified and their radiochemical purity established by isotopic dilution analysis. [7-3H]16alpha-hydroxy-dehydroepiandrosterone (2.5 x 10(7) dpm) was obtained by microbial hydroxylation of substrate (1.9 X 10(9) dpm). In some cases [7-3H])5-androstene-3beta, 16alpha, 17beta-triol in a small amount of radioactivity could be found at the prolonged reaction for 30 hr.
 
Article
The introduction of a 16 alpha-hydroxyl function into the steroid nucleus was studied in resting cells of Streptomyces roseochromogenes NRRL B-1233. The oxidation product of dehydroepiandrosterone (DHEA) was identified as 16 alpha-hydroxy DHEA by using thin-layer and gas-liquid chromatography. A linear relation between cell concentration and 16 alpha-OH-DHEA formation was observed. 16 alpha-Hydroxylase showed good activity at pH 8.0 for 16 alpha-OH-DHEA formation. The enzyme showed good activity at 3.1 x 10(-4) M DHEA. The oxidation products of pregnenolone, 4-androstene-3,17-dione, estrone, and 5-androstene-3 beta,17 beta-diol as well as of other substrates were identified as the 16 alpha-hydroxy steroid, respectively. The rates of microbial 16 alpha-hydroxylation were as follows: 76.9% for DHEA, 50.4% for pregnenolone, 43.9% for 4-androstene-3,17-dione, 34.3% for estrone, and 19.6% for 5-androstene-3 beta,17 beta-diol. The organism tested catalyzes 16 alpha-hydroxylation of a wide variety of steroids.
 
Article
The comparative fatty acid analysis of extractable and non-extractable lipids of Escherichia coli W 1655 F+ and its stable protoplast type L-form shows quantitative as well as qualitative differences. From 10 different fatty acids obtained 16:0, 17:0 and 18:0 are present at about the same quantities in the lipid fractions of the bacterial and L-form. The absence of larger amounts of 12:0, 14:0, and 14:beta OH fatty acids in non-extractable L-form lipids reflects the loss of the cell wall in L-form cells. 16:1 fatty acid was found in L-form lipids only. This qualitative difference and the 2-3 times higher content of 18:1 in L-form lipids and the 7 times lower content of cyc 19:0 in extractable lipids of the L-form may be interpreted as alterations characteristic for the changed composition of the cytoplasmic membrane in L-form cells.
 
Article
The steroid 16alpha-hydroxylase of Streptomyces olivoviridis was shown to be constitutive, but synthetized only in small amounts in the absence of cortisol. The induction of the enzyme with the steroidal substrate resulted in a distinct increase in the 16alpha-hydroxylation rate. This increase in activity, however, could be demonstrated only in later phases of culturing, after the growth of the mycelium was completed. The possible mechanisms controlling the activity of the enzyme during the growth phase of the culture are discussed.
 
Article
Streptomyces roseochromogenes (NRRL B-1233) converted estrone predominantly into its 16 alpha-hydroxyl derivative. Chemical and spectroscopic (UV, IR, NMR, MS) methods were used in establishing the structure and strereochemistry of the oxidation product. The product was assigned as 16 alpha-hydroxyestrone (yield, 17%). No other oxidation product was detected in this experiment. An interrelationship between cell growth and 16 alpha-hydroxy-estrone formation was observed. Also, 16 alpha-hydroxylation of estrone was observed in resting cells. 16 alpha-Hydroxylase showed good activity at 3.7 X 10(-4)M of estrone concentration and was completely inhibited by 1.1 X 10(-3)M. These results indicate the presence of a constitutive 16 alpha-hydroxylase in the organism investigated.
 
Article
After cultivation of Streptomyces hydrogenans in the presence of different steroids the activity of both 3 alpha, 20 beta-hydroxysteroid dehydrogenase and 3 beta, 17 beta-hydroxysteroid dehydrogenase was determined in the cell homogenate of the microorganism. By comparing the efficacy of the steroids to increase enzyme activities, steroids could be divided into 3 groups: a) steroids which stimulated preferentially the activity of 3 alpha, 20 beta-hydroxysteroid dehydrogenase (e. g., corticosterone), b) steroids which stimulated preferentially 3 beta, 17 beta-hydroxysteroid dehydrogenase (estradiol-17 beta), and c) those behaving intermediately (e. g., progesterone, 5 alpha -dihydrotestosterone). Highest 3 beta, 17 beta-hydroxysteroid dehydrogenase activity could be measured 2 h after addition of 5 alpha-dihydrotestosterone to the culture medium. The activity of 3 alpha, 20 beta-hydroxysteroid dehydrogenase, however, increased continuously up to 4 h. 3 alpha, 20 beta-hydroxysteroid dehydrogenase and 3 beta, 17 beta-hydroxysteroid dehydrogenase syntheses seemed to be controlled by steroids in a non-coordinate manner.
 
Article
The antibacterial activity of ten N-alkylated derivatives of daunorubicin and adriamycin as well as of 5-iminodaunorubicin has been tested by using Bacillus subtilis 170, Escherichia coli B, and Proteus mirabilis VI and their stable protoplast type L-forms in an agar diffusion test. Eight of the substances showed similar activities against B. subtilis and the L-forms of all test organisms, but no activity against the bacterial forms of E. coli and P. mirabilis. The cell wall of these gram-negative bacteria is responsible for this resistance by not allowing the antibiotics to enter the cells. The piperidino compound N-(CH2)5 daunorubicin shows 2-4 times higher activity against B. subtilis and all L-forms in comparison to daunorubicin and the other derivatives. Five of the substances were inactive against all test strains. Their inactivity seems to be associated with the larger substituents at the C-3' position. Relations between molecular structure and activity are discussed considering data about the interaction with DNA and the antitumor activity. Stable protoplast type L-forms and their bacterial forms represent a suitable and effective test system to screen for more effective substances and to get more information about their mode of action.
 
Article
The dap-dependent mutant of Escherichia coli could grow in the presence of several analogues of diaminopimelic acid (dap). The multiplication proceeded without any apparent difference for several hours when dap was replaced with 2,6-diamino-4-fluoro-pimelic acid or lanthionine. Only growth without accompanying division could be detected in the presence of 2,6-diamino-4-hydroxypimelic acid (hydroxy-dap). 3H-hydroxy-dap was incorporated into the TCA precipitable material under these conditions. 2,4,6-triaminopimelic acid, probably also 2,6-diamino-5-oxo-pimelic acid could not substitute dap in the synthesis of murein. The synthesis of DNA measured by the incorporation of 3H-thymidine was not inhibited by hydroxy-dap but the formation of the cross wall was impaired. The rods were longer, formed bulges in the central part of the cell and ultimately lyzed. Selenium-lanththionine was more toxic but even in its presence some growth occurred and DNA synthesis took place. The replacement of dap with hydroxy-dap stimulated the degradation of the wall mucopeptide during the growth.
 
Top-cited authors
Emi Kleiner
  • Interdisciplinary Center Herzliya
Peter Biely
  • Slovak Academy of Sciences
Diego Matteuzzi
  • University of Bologna
Olli H. Tuovinen
  • The Ohio State University
Rayane Bode
  • Istanbul Ticaret University