The plasmid genes encoding the ability to grow chemolithoautotrophically with H2 and CO2 from a H2-oxidizing bacterium Alcaligenes hydrogenophilus were in vivo cloned using the broad host range Inc P1 R-plasmid R68.45. The genes of H2-oxidation enzymes were expressed by a novel alternative σ54-like factor encoded on the chromosome. The σ54-like factor was also in vivo cloned using R68.45. Both R68.45-primes, one carrying plasmid genes and the other carrying chromosome genes, were in vivo recombinated and a recombinant plasmid carrying both genes from plasmid and chromosome was obtained. The conjugal transfer of chemolithoautotrophically growing ability was carried out using the resulting recombinant plasmid. Seventeen bacterial strains, including useful material-producing bacteria, grew up to be able to grow with H2 and CO2 as the H2-oxidizing bacteria. Some patent strains registered for the production of antibiotics were ascertained to produce some products which inhibited the growth of the testingbacteria, under not only heterotrophic conditions but also chemolithoautotrophilc conditions. The results obtained in our studies will be available in the future research for the production of useful material from CO2.
New anti-rust quaternary ammonium salts such as N-alkyl-N-(2-hydroxy-3-phenoxy) propyl-N, N-dimethylammonium butyl phosphate (5) [alkyl homologues are octyl (5a), decyl (5b), dodecyl (5c), tetradecyl (5d), hexadecyl (5e), and octadecyl (5f)] were synthesized in order to find the relationship between the alkyl length and the antibacterial activities. N-Alkyl-N-(2-hydroxy-3-phenoxy) propyl-N, N-dimethylammonium chloride (6) [alkyl homologues are dodecyl (6c), tetradecyl (6d), hexadecyl (6e)] were used as standard compounds in the quantitative analysis. 5c and 6c showed antibacterial activities and these activities were nearly equal to that of benzalkonium chloride (BAC), but, the activities shown by 5e, 5f and 6e were insufficient. 5c-e, showed excellent anti-rust effects, but 6c-e showed insufficient effects, because the former had a butyl phosphate anion and the latter had a chloride anion as a counter ion. 5a, b were insoluble in water. 5c showed a cloud point at 61.5°C and 5d showed a cloud point at 72.0°C at a 10% concentration in water. This phenomenon is similar to those of non-ionic surfactants. 5e was freely soluble in water.
Crystal morphology and habit analyses necessary for a pharmaceutical technique have been studied by a polarizing microscope for tiny crystals of sulfamethizole, levodopa and phenobarbital listed in the Japanese Pharmacopoeia XII. Measurement of key refractive indices by an immersion method has been tried semiautomatically using photographs and the kit of immersion oils produced at 0.005 intervals, and a decision of an orientation of the section has been also carried by conoscope using photos. From the orthographic projections produced by the habit analysis of the colored photo as mentioned in the previous reports morphological and optical properties were clarified.
The antioxidative effect of the mammalian liver hydrolyzate preparation (LH) was investigated mainly in the mouse liver homogenates. At lower concentrations, LH stimulated lipid peroxidation induced by Fe2+ and ascorbic acid in the homogenates, but inhibited completely the oxidation at higher concentrations. LH also inhibited the peroxidation of linolenic acid and linoleic acid. LH reduced 1, 1-diphenyl-2-picrylhydrazyl, a free radical model, in a concentration-dependent manner, suggesting that the radical scavenging property of LH is involved in its antioxidative action. Fe2+ and glutathione, contained in LH, seem to act as a stimulator and an inhibitor of peroxidation, respectively.
In order to examine analgesic and antiinflammatory activities of the position isomers of 4-ethoxy-2-methyl-5-morpholino-3 (2H)-pyridazinone (1, emorfazone), an analgesic-antiinflammatory drug, 5-ethoxy-2-methyl-4-morpholino-3 (2H)-pyridazinone (2), 6-ethoxy-2-methyl-4-morpholino-3 (2H)-pyridazinone (3) and 6-ethoxy-2-methyl-5-morpholino-3 (2H)-pyridazinone (4) were prepared. Since 4 showed the most strong activity among the compounds tested, various 6-alkoxy- or 2-cyclohexyl- or 2-phenyl-5-substituted amino-3 (2H)-pyridazinones (15, 16) were prepared and examined for their activities. As a result, 4 and 2-methyl-5-morpholino-6-n-propoxy-or 6-n-butoxy-3 (2H)-pyridazinone (15b, c) and 6-ethoxy-2-ethyl-5-morpholino-3 (2H)-pyridazinone (151) were revealed to be more potent in analgesic and antipyretic activities than commercial drugs (1, aminopyrine, mepirizole, tiaramide HCl, phenylbutazone, mefenamic acid). On the basis of the available data, the structure-activity relationship in a series of 6-alkoxy-2-alkyl-5-substituted amino-3 (2H)-pyridazinones (15, 16) was also discussed.
Stereochemical studies on palladium-catalyzed asymmetric sulfonylations of α, γ-disubstituted allylic acetates were carried out using several kinds of chiral phosphine ligands under various reaction conditions for the development of new methodologies of chiral carbon-sulfur bond formations.
Deoxyribonucleases from eggs and the liver of Xenopus laevis were partially purified by DEAE-cellulose and heparin-Sepharose affinity column chromatographies. The fractions having egg and liver DNase activities were eluted on high performance liquid chromatography through TSK gel G3000SW at the molecular weights of 41.5 and 45 kDa, respectively. The frog DNases hydrolyzed a native DNA over a heat-denatured DNA, and also formed double-strand cuts not only in linear λ-DNA but also in closed circular pBR322DNA. The pH optimum of the DNases was 4.5-5.0 in 50 mM acetate buffer. These enzyme activities were abolished by treatment at 80°C for 5 min and pH 2, 3 or 12 for 1h. The enzymes act in such a manner as deoxyribonuclease II (from bovine spleen)-type nuclease with respect to substrate specificity, optimum pH and cation dependence.
In the course of our search for natural antioxidants, the methanol extract of the fruit hulls of mangosteen (Garcinia mangostana L.) originating in Vietnam was found to exhibit a potent radical scavenging effect. By monitoring the radical scavenging effect, two xanthones, α- and γ-mangostins, were isolated together with (-)-epicatechin, procyanidins A-2 and B-2 as active principles. The antioxidative activity of these two xanthones was measured by the ferric thiocyanate method and it was found that γ-mangostin showed more potent antioxidative activity than BHA and α-tocopherol.