Metabolisms of Nucleosides in Bacteria

In this study, the authors developed a simplified method for the separation and the quantitative determination of nucleosides and bases, using paper electraophoretic technique. By this method, nucleosides and bases were well separated and determined in a fairly short time. Thus, this method was expected to be as accurate as the published methods and was believed to be a better method for both quantitative determination and detection of the nucleosides and bases in a large number of samples.     

Structural Requirements for Mutagenic Activities of N-Heterocyclic Bases in the Salmonella Test System

The mutagenic activities of quinoline, isoquinoline, phenanthridine, benzo(f)quinoline, benzo(h)quinoline and their α-amino derivatives were compared in relation to the effect of structural changes using the Salmonella typhimurium test system. All mutagenic compounds tested require the liver microsomal fraction for their mutagenic activity. Phenanthridine, two benzoquinolines and quinoline were mutagenic. α-Amination of two benzoquinolines and quinoline resulted to increase their mutagenic activity intensively. Addition of a benzene ring to the benzene moiety of 2-aminoquinoline, so that two carbon atoms are shared, affected distinctly the increase in the mutagenic activity. The co-existence of benzoquinoline series with 2-aminobenzo(f)quinoline showed the clear synergistic action.     

Microbial Oxidation of Cephalosporins to Cephalosporin Sulfoxides

The highest inhibition rate of conidial germination of Pyricularia oryzae was shown by extracts of rice plant leaves inoculated by a pathogen after treatment with probenazole, a rice blast controlling agent. Four anti-conidial germination substances were isolated from these extracts. Substances A, C and D inhibited the germination of the conidia at concentrations between 100 and 200 mcg/ml, and substance B caused morphological changes in the germination tubes of the conidia with a little inhibition of germination. These substances were differentiated from momilactone A, B and the degraded or metabolized products of probenazole. Besides anti-conidial germination activity, they showed antimicrobial activities against several kinds of phytopathogenic bacteria of fungi on agar plates by diffusion method.     

A New Isoflavone with Antifungal Activity from Immature Fruits of Lupinus luteus

A new isoflavone having antifungal activity was isolated from immature fruits of Lupinus luteus (Leguminosae), and named luteone. The structure was shown to be 5,7,2′,4′-tetrahy-droxy-6-(3,3-dimethylallyl)-isoflavone by degradative and spectroscopic studies.     

Studies on Microbial Metabolisms of Sugar Nucleotides

Effects of various factors including incubation time, water content of airdried cells, concentration and pH of KH₂PO₄–K₂HPO₄ mixture, d-glucose concentration, MgSO₄ concentration, GMP concentration, cell concentration, aeration and various kinds of carbohydrates on the fermentative production of GDP-mannose, GDP and GTP from 5′-GMP by air-dried cells of baker’s yeast were investigated. The water content of air-dried cells was the most important factor in the fermentation. When the air-dried cells of baker’s yeast (100 mg/ml) were incubated with 5′-GMP (20 μmoles/ml), d-glucose (800 μmoles/ml), potassium phosphate buffer (360 μmoles/ml, pH 7.0), and MgSO₄ (20 μmoles/ml), 2-hr incubation gave GDP in 20% yield and GTP in 61.1% yield, GDP-mannose being produced in 45% yield after 8-hr incubation. The phosphorylation of 5′-AMP, 5′-dAMP, 5′-dGMP 5′-CMP and 5′-UMP was also observed in high yields under the same conditions.     

Microbial Formation of D-Pantothenic Acid-α-glucoside

The formation of D-pantothenic acid-α-glucoside (PaA-α-G) was found from D-pantothenic acid (PaA) and maltose in incubation mixtures of microorganisms, especially Saccharomyces yeasts and Sporobolomyces coralliformis IFO 1032. The reaction conditions were investigated for formation of PaA-α-G by resting cells of Spor. coralliformis. The formation of the compound increased with PaA concentration (3~20 mg/ml). The yield was maximum at 5~10 mg/ml of PaA. Cetyl trimethyl ammonium bromide (0.1 %) promoted the formation of PaA-α-G. Sucrose was the optimal α-glucosyl donor. When 30 mg/ml of sucrose was fed to the reaction mixture (initial sucrose, 100 mg/ml; and PaA, 10 mg/ml) at 12-hr intervals, 5.74 mg/ml (3.30 mg/ml as PaA) of PaA-α-G was formed in 48-hr incubation at 28°C with shaking. PaA-α-G was also formed by yeast α-glucosidase, mold maltase and the cell-free extract of Spor. coralliformis. The compound showed approximately 9~10% and 0.1~0.3% (molar ratio) of activity of PaA for Saccharomyces carlsbergensis ATCC 9080 and Lactobacillus plantarum ATCC 8014, respectively. The compound had the same microbiological activity as authentic 4′-O-(α-D-glucopyranosyl)-D-pantothenic acid.     

Acid-stable α-Amylase of Black Aspergilli

The Acid-stable α-amylase and the acid-unstable α-amylase from Aspergillus niger contained one mole of sulfhydryl group per one mole of enzyme, which probably existed correlating with calcium atom that was essential for the amylase activity.

Iodine reacted at acidic pH specifically with the sulfhydryl group of both enzymes and oxidized it to considerably high degree, since about 4 eq of iodine per mole of sulfhydryl group of both enzymes were consumed. The modification of the sulfhydryl group of the acid-stable α-amylase did not affect the amylase acitvity, while, that of the acid-unstable α-amylase reduced it to 70 per cents intact enzyme. It was difficult to carry out carboxy-methylation of the sulfhydryl group of the acid-stable α-amylase under mild conditions maintaining its activity, but that of the acid-unstable α-amylase was easily achieved.

These facts suggested that some differences existed in the neighborhood of the sulfhydryl group of both enzymes, and that the sulfhydryl group of them was not the active site.     

The Influence of Measurement Parameters on the Specific Rotation of Amino Acids

The effects of solute and hydrochloric acid concentrations on optical rotation were studied using 20 naturally occuring amino acids.

There appeared to be no common factor among the amino acids as far as the inclination of optical rotation was concerned. Lutz-Jirgenson’s rule could be applied to few amino acids in the cationic form. Therefore, in the determination of the optical rotation, the concentration of the solute, nature of solvent and temperature must be rigorously controlled. The optical conditions of measurement and the specific rotation of 20 amino acids were recommended based on this work.     

Formation of Rhamnolipid by Pseudomonas aeruginosa and its Function in Hydrocarbon Fermentation

Screening test for obtaining growth stimulant (GS) produced by a hydrocarbon-utilizing bacterium, Pseudomonas aeruginosa S₇B₁, was carried out. In consequence, the anthrone positive substance was most effective on the growth of this strain. Although the growth of this strain on glucose medium had no relation with the addition of GS, the growth on n-hexadecane medium was remarkably stimulated by the addition of GS. This effect of GS seemed to be specific on the growth of P. aeruginosa. GS which had a strong surface activity and emulsifying power was comfirmed to be rhamnolipid.