Penicillinamidohydrolase inEscherichia coli

Substrate specificity of the bacterial penicillinamidohydrolase (penicillinacylase, EC 3.5.1.11) fromEscherichia coli was determined by measuring initial rates of enzyme hydrolysis of different substrates within zero order kinetics. SomeN-phenylacetyl derivatives of amino acids and amides of phenylacetic acid and phenoxyacetic acid of different substituted amides of these acids or amides, structurally and chemically similar to these compounds, served as substrates. Significant differences in ratios of initial Tates of the enzyme hydrolysis of different substrates were found when using a toluenized suspension of bacterial cells or a crude enzyme preparation, in spite of the fact that the enzyme is localized between the cell wall and cytoplasmic membrane, in the so-called periplasmic space.N-phenylacetyl derivatives are the most rapidly hydrolyzed substrates. Beta-phenylpropionamide and 4-phenylbutyramide were not utilized as substrates. The substrate specificity of the enzyme is discussed with respect to a possible use of certain colourless compounds as substrates, hydrolysis of which yields chromophor products suitable for a simple and rapid assay of the enzyme activity.     

Reversion of anomalous forms ofBacillus subtilis

Anomalous forms ofBacillus subtilis A 32 produced by prolonged cultivation in a chemostat under nitrogen limitation are described. A change in the cultivation conditions brings about a transformation of these forms to bacillar rods. The transformation is gradual and lasts for several generations.     

Biochemical dehydrogenations of saccharides

Fermentation of a medium containing 5% 2-deoxy-D-glucose and barium carbonate by a strain ofPseudomonas aeruginosa yielded barium 2-deoxy-d-gluconate. The yield was 77% theoretical. The strain in question makes it possible to prepare directly calcium, magnesium, manganese and ferrous salts of 2-deoxy-d-glueonic acid. A treatment of 6% solution of 2-deoxy-d-glucose with commercial glucose oxidase preparation caused also a complete dehydrogenation.     

A study of the binding of Mn2+ to bovine pancreatic deoxyribonuclease I and to deoxyribonucleic acid by electron paramagnetic resonance.

EPR studies of Mn2+ binding to bovine pancreatic deoxyribonuclease I show that the enzyme can bind three Mn2+ ions at pH 7.5 and 2 degrees. Two sites bind Mn2+ strongly, with a Kd of 10(-4)M, and the third binds Mn2+ weakly, with a Kd of 10(-3)M. Ca2+ competes with the two strong sites, whereas Mg2+ competes only with one of them, indicating that both sites are not equivalent. Mn2+ binding to DNA has been confirmed by EPR measurements. Two types of sites, with different affinities for Mn2+ binding, were found on DNA molecules, one with a Kd of 1.2 times 10(-4)M and the other with a Kd of 10(-3)M. Mg2+ ions can displace Mn2+ from the high affinity sites, but not from the low affinity sites. These results suggest the Mn2+ binds not only to the phosphate groups, but also to the electron donor groups of the base rings.     

Ultrastructure of rabbit lung intravenously infected with Mycobacterium simiae "Weiszfeiler" no 52.

M. simiae "Weiszfeiler" strain No. 52 belongs to the second Runyon Group of Atypical Mycobacteria characterized by rough, eugonic, yellow-orange pellicle grown on Sauton medium incubated for 14 days at 37 degrees C. A suspension containing 10 mg/ml (equalling 5 times 10-7 viable units) injected intraveneously into rabbits provokes profound changes in the ultrastructure of the pulmonary tissue, manifested in activation of macrophages and pathologic alteration of pneumocytes. In histologic sections stained according to the Ziehl-Neelsen technique and examined in a light-microscope cytoplasmic acid-fast, highly dispersed granules are found in macrophages, resembling in morphological appearance the cellular phospholipids in sarcoid granuloma. Abundant production of interalveolar and interseptal fibrotic collagen tissue was observed in rabbits infected with M. Simiae a rather striking phenomenon in fresh experimental mycobacterial infection in the rabbit. The strain in question should therefore be considered not as Saprophyte but as a Mycobacterium Species with attenuated virulence, closely related to Prissic and Masson's M. scrophulaceum.     

Diabetogenic effect of triamcinolone in man with impaired thyroid function: serum free fatty acids, inorganic phosphorus, calcium and total protein pattern after an oral glucose load.

In a group of 35 subjects (12 hypothyroid, 11 euthyroid and 12 hyperthyroid) the changes of FFA, inorganic phosphorus, calcium and total proteins were investigated during glucose tolerance test with (TGTT) or without (OGTT) triamcinolone pretreatment. In hypothyroidism the decrease of FFA during TGTT was blunted, whereas the pattern of phosphatemia was unaltered and total proteins were increased. In euthyroidism triamcinolone exerts the opposite effect on the behaviour of FFA (decline was smaller and shorter) and phosphatemia (decline was pronounced and persists for longer period). In hyperthyroidism the fasting level of FFA decreased in TGTT but during the test no significant differences in FFA and phosphatemia patterns were observed. The calcemia was not influenced by triamcinolone in any subgroup. Our results suggest the importance of thyroid function as modifying factor in some metabolic effects of triamcinolone in man. Changes in hypothyroidism are in good agreement with changes of blood glucose and insulinemia, as described previously. Opposite effect of triamcinolone on FFA and phosphatemia pattern in euthyroidism may suggest the changes of sensitivity of some peripheral tissues to insulin action. The decline of fasting level of FFA after triamcinolone in hyperthyroidism is surprising and might be of importance in maintaining unaltered glucose tolerance after triamcinolone in this subgroup.     

Physiological characteristics of chemostatically grownCitrobacter freundii as a function of the specific growth rate and type of nutrient limitation

Citrobacter freundii was grown aerobically in a chemostat on a mineral medium witn galactose or glucose as carbon and energy sources under limitation by carbon or nitrogen source respectively. At various specific growth rates ranging from 7 to 95% μ_max the culture in steady state was analysed and growth yield, specific metabolic rate of substrate utilization, intracellular concentration of pyruvate, ATP, ADP, AMP and energy charge were determined and plotted as functions of dilution rate. In all four types of experiments the physiological state of cells remained practically independent of dilution rate up toD = 0.6 μ_max, and at a given specific growth rate nearly independent on μ_max and type of limitation. At approximatelyD = 0.6 μ_max, which is close to the maximum output dilution rateD _m, the physiological state of the cells changed: growth yields decreased and intr cellular pyruvate and adenylates concentrations increased. Consequently, in a given medium two dilution rates exist at which growth rate dx/dt is the same but the physiology of the population is quite different.     

Oxidation of methanol,formaldehyde and formic acid by methanol-utilizing yeast

Methanol-utilizing yeast,Candida boidini 11 Bh, characterized by high tolerance to methanol during growth, displays even higher tolerance when the oxidation rate by intact cells is tested. Low respiration activity is found even at 22% v/v of methanol. The half-saturation constant was 17–18mM. The half-saturation constants for the two oxidation intermediates, formaldehyde and formic acid were 3.6–4.0 and 30–33mM, respectively. When applied together with standard concentration of methanol, very low concentrations of both intermediates stimulated the oxidation rate. These results are discussed in connection with the relationship between growth and oxidation, the tolerance to high concentrations of inhibitory products and the mechanism of inhibition.     

Viererbruten des Kaiseradlers(Aquila heliaca)

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