Comparison of Mineral Balances in Germfree and Conventional Mice When Sodium Phytate is Added to Purified Diet

A strain of Alcaligenes isolated from soil was a good producer of β-glucuronidase, and the enzyme was purified from the cell-free extract by sequential column chromatography on DEAE-Toyopearl, Toyopearl HW-55F, and Phenyl-Sepharose CL-4B. By these procedures, two β-glucuronidases designated as β-glucuronidases I and II were purified 240- and 508-fold, respectively. β-Glucuronidase I, with a molecular weight of 75,000, had an optimum pH at 7.5 and the enzyme II, with a molecular weight of 300,000, had maximum activity at pH 6.0. Both enzymes were strongly inhibited by saccharo-1,4-lactone, glucaro-δ-lactam, p-chloromercuribenzoate, Hg²⁺, and N-bromosuccinimide. β-Glucuronidase I was active toward estrogen-3-β-glucuronides and inert toward β-glucuronide conjugates of menthol, estrogen-17β-, estrogen-16α-, androsterone-3α-, testosterone-17β-, cortisol-17α-. β-Glucuronidase II hydrolyzed all of these substrates. β-Glucuronidase I was inhibited by phenolphthalein and its glucuronide.     

Structures of Allosamidins,Novel Insect Chitinase Inhibitors,Produced by Actinomycetes

The structures of allosamidin (1) and methylallosamidin (2), novel insect chitinase inhibitors, were elucidated as 1 and 2 by acid hydrolysis experiments and analyses of 2d-NMR spectra. They are unique basic pseudotrisaccharides consisting of 2-acetamido-2-deoxy-d-allose (N-acetyl-d- allosamine) and a novel aminocyclitol derivative (3), termed allosamizoline.     

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.     

The Chlorinolysis of Methionine Derivatives

The chlorinolysis of l-methionine methyl ester hydrochloride with molecular chlorine was carried out under various conditions, resulting in methyl l-2-amino-4,4,4-trichlorobutanoate and methyl l-2-amino-3,4,4,4-tetrachlorobutanoate which were isolated as N-benzoyl and N-carbobenzoxy derivatives. The chlorinolysis of N-acylmethionine ester and methionine sulfoxide ester proceeded also without cleavage of the N-protecting group to give the same products as above. However, the reaction of methionine sulfone derivative with chlorine did not proceed in the same conditions.

It was proved that the resulting polychloroamino acid derivatives are optically pure. The possible chlorinolysis mechanism was also proposed.     

Syntheses and Biological Activities of Isonitrile Dipeptides

We investigated GroEL substrates from Bacillus subtilis 168 using the single-ring mutant of B. subtilis GroEL. We identified 28 candidates for GroEL substrates, of which Spo0B, Ald, Eno, SpoIIP, and FbaA were involved in spore formation, and Rnc, Tuf, Eno, Tsf, and FbaA were essential for B. subtilis growth. As observed at the protein level, the amount of SpoIIP interaction with GroEL increased at 3 h after initiation of sporulation.     

Isolation and Identification of Ubiquinone 10 from Cultured Cells of Tobacco

Callus tissues were induced from stem and root segments of Rauwolfia serpentina. Growth and alkaloid production of the callus tissues were examined under various culture conditions. The growth was strikingly promoted in the presence of 2,4-D (0.5~1 ppm), kinetin (0.2~0.5 ppm) and yeast extract (0.1~0.2%). At favourable conditions, the growth value in 4 weeks’ culture was ca. 40 (F.W.), and ca. 25 (D.W.) for stem callus tissues, and ca. 15 (F.W.), and ca. 8 (D.W.) for root callus tissues. Stem and root callus tissues produced ajmaline and some other unidentified Rauwolfia alkaloids. The ajmaline content in root callus tissues was 10~20mg % and in stem callus tissues was 1~10mg %. The ajmaline production was strikingly reduced when 2,4-D concentration increased, or kinetin was omitted in the culture medium. Phytosterols including stigmasterol, β-sitosterol or cholesterol were also produced.     

Isolation and Structure of Phytotoxic Compounds Produced by Phyllosticta sp.

In the course of phytopathological studies, it has been observed that phyllosticta sp. produces a toxic compound causing wilting and simultaneous dark coloration of the clover leaf. A toxin, herein refered to as phyllosinol (mp 76~77°C) was isolated in a crystalline state from the pure culture and it was proved to be the principal toxic metabolite of the fungus. Although the melting point of phyllosinol differed significantly from that reported for epoxydon (40~45°C), the structural evidence deduced from spectrometric and chemical methods indicates that phyllosinol is substantially identical with epoxydon even in stereo-isomeric considerations. In the red clover leaf test 10 ppm phyllosinol showed the wilting effect.     

Growth Profile of Crown Gall Cells of Tobacco in Suspension Culture

In order to obtain a basic information of plant cell suspension culture as a step toward the development of large scale culture, culture conditions of crown gall cells (auxin non-requiring cells) were investigated. Addition of yeast extract to culture medium was significantly effective for the growth and cell dispersion.

In experiments on the ability of the cultured cells to utilize sugars as the carbon source, it was observed that galactose, added to the culture medium, markedly inhibited the cell growth.

Pasteurization of the medium containing fructose as carbon source made it brownish by Maillard reaction and the medium apparently restrained the cell growth. However, the fructose medium sterilized by filtration was excellent for the cell growth as well as sucrose or glucose medium. In a jar fermentor, even the glucose medium became brownish by heat sterilization and the brown colored medium restrained the cell growth. Under optimum conditions, the doubling time was 1.1 day in exponential phase and 2.0 g of cell (dry weight) per 100 ml culture was obtained as the maximum yield.