An amylase inhibitor-producing microorganism was identified as a subspecies of Streptomyces diastaticus from morphological and physiological studies and was named Streptomyces diastaticus subsp. amylostaticus No. 2476.
When this strain was aerobically cultured in a shaking flask containing 100ml of medium consisting of 4% corn starch, 2% soy bean flake extract, 0.3% NaCl, 0.1% K₂HPO₄, 0.05% MgSO₄•7H₂O, 0.001% FeSO₄•7H₂O, 0.0001% CuSO₄•5H₂O, 0.0001% ZnSO₄•7H₂O, and 0.0001% MnSO₄•nH₂O (pH 7.0) at 30°C, the highest inhibitory activity was obtained after 70_??_80hr of cultivation.
This amylase inhibitor (S-AI) had inhibitory activity on α-amylases and glucoamylase, but not on β-amylases and pullulanase.

View full abstract

Degradation of eugenol as the sole carbon and energy source was studied with a newly isolated microorganism. Ferulic acid (4-hydroxy-3-methoxycinnamic acid) and vanillin (4-hydroxy-3-methoxybenzaldehyde) were obtained as products. These are produced by oxidation and β-oxidation of the aromatic side-chain and metabolized further to form vanillic acid and protocatechuic acid. Protocatechuic acid was cleaved by “ortho” fission to form β-ketoadipate. From these results, the degradation pathway was proposed. The strain was tentatively identified as Corynebacterium sp. from the results of physiological and morphological tests.

View full abstract

The inhibitory action of lauryl DL-valinate•HCl on plant growth was studied with rice (Oryza sativa L. cultivar. Nihonbare), millet (Panicum crus-galli) and barnyardgrass (Echinochloa crus-galli). It has been found that the inhibitory activity of amino acid higher alkyl esters including lauryl DL-valinate•HCl on plant growth was closely related to their wetting power. The leakage of protein from seeds submerged in the solution of lauryl DL-valinate•HCl was observed to be proportional to its growth-inhibiting activity. Lauryl DL-valinate•HCl inhibited the α-naphthylamine-oxidizing activity of roots and the α-amylase synthesis induced by gibberellin in isolated endosperms.

View full abstract

An α-1, 3-glucanase was partially purified from Meicelase, a commercial cellulase preparation from T. viride, and its properties were studied. Cellulase in the crude enzyme was removed by adsorption on cellulose powder, and α-1, 3-glucanase was purified through saltingout with ammonium sulfate, chromatographies on DEAE-Sephadex, hydroxylapatite and Sephadex G-100. The α-1, 3-glucanase gave 2 peaks on DEAE-Sephadex chromatography, and the enzyme of the second peak was further purified to about 300-fold purification. The enzyme was most active at pH 4.5, and 55°C, stable at 4°C at pH between 3 and 7, and stable up to 50°C on heat treatment at pH 4.5 for 30min. The Michaelis constant for α-1, 3-glucan was 7.1mM (as anhydroglucose units). The molecular weight of the enzyme determined by gel filtration on Sephadex G-100 was 47, 000. Only glucose was formed as reducing sugar liberated from α-1, 3-glucan, showing that the enzyme was an exo-α-1, 3-glucanase (EC 3. 2. 1. 84).

View full abstract

When Rhizopus arrhizus NRRL 1526 was mix-cultured with Proteus vulgaris AHU 1144, a strain having a high fumarase activity, in a medium containing glucose as a substrate, fumaric acid fermentation was successively converted to L-malic acid fermentation and large amounts of L-malic acid were accumulated as an end product.
As an inoculum of P. vulgaris for this fermentation, cells in the stationary growth phase (48 to 72 hr culture) were much more favorable than those in the exponential growth phase (18 hr culture) and malic acid yields in the former case were as high as about 70 to 75% based on initial glucose after 3 to 4 days of the mixed culture.

View full abstract

Gamma-radiolosis of D-glucose in aqueous solution under anaerobic condition was investigated. D-Glucose was significantly decomposed by action of a hydroxyl radical, a primary radiolytic product of water. Under anaerobic condition, glucose radicals formed by hydrogen abstraction of hydroxyl radical were dehydrated to give deoxygenated sugars, and a part of the radicals followed disproportionation reaction to afford dicarbonyl sugars. Among the products formed through these reactions, 2-deoxy-D-arabinohexono-1, 4-lactone (G in N₂=0.6, G in N₂O=0.8), 3-deoxy-D-erythrohexosulose and other 3-deoxy sugars as deoxygenated compounds, and D-arabinohexosulose and D-xylohexos-5-ulose as dicarbonyl compounds were identified. From these results, the radiolytic scheme of D-glucose in the absence of oxygen was proposed.

View full abstract

Guanidinoacetate amidinohydrolase was purified about 45 fold and crystallized from Pseudomonas sp. ATCC 14676 grown in a medium supplemented with guanidinoacetate. The molecular weight of the enzyme was estimated to be about 163, 000 by two different procedures. Polyacrylamide disc electrophoresis of the enzyme in the presence of SDS showed a single component with a molecular weight of 38, 000, suggesting that the enzyme is composed of four identical subunits. Treatment of the enzyme with EDTA resulted in the complete loss in the activity, which was fully restored by incubation with Mn²⁺. A slight activation was observed with Fe²⁺, Co²⁺, and Ni²⁺ (0.1mM), while these cations added at 1.0mM together with 0.1mM Mn²⁺ completely inhibited the enzyme. The enzyme was optimally active at pH 9.0 to 9.5. Guanidinoacetate was the exclusive substrate among the guanidino compounds tested. The apparent Km values for guanidinoacetate and Mn²⁺ were estimated to be 9.1mM and 1.3 μm, respectively. The enzyme was activated by incubation with 2-mercaptoethanol added together with Mn²⁺ and was inhibited by p-chloromercuribenzoate; indicating that the presence of sulfhydryl groups at or near the active site of the enzyme. ATP inhibited the enzyme and the inhibition was competitive with Mn²⁺.

View full abstract

Candida brumptii IFO-0731, which is known to produce succinic acid from n-paraffins, also produced other organic acids. One of them was identified as L-malic acid. Some culture conditions for L-malic acid production from n-paraffin were investigated. It was found that urea was the most favorable nitrogen source and 2g/liter of urea was optimum for L-malic acid production. Under the best condition, 25.4g/liter of L-malic acid was obtained from 4% (v/v) of super heavy n-paraffin and the yield was about 80% (wt./wt.).

View full abstract

A strain of Erwinia aroideae produced an extracellular pectolytic enzyme under growth conditions with pectin or pectic acid as the inducer. This strain also produced a pectin lyase when nalidixic acid is added to a culture medium. The pectolytic enzyme produced under the growth conditions was purified approximately 40-fold from the culture fluid by carboxymethyl cellulose and Sephadex G-75 gel column chromatographies. The purified enzyme was almost homogeneous on sodium dodecyl sulfate polyacrylamide gel electrophoresis, having a molecular weight of about 36, 000 to 38, 000. This enzyme, with optimal activity at pH 9.0 to 9.2, produced reaction products which had a strong absorption at 230 nm, indicating a lyase type of the reaction. The enzyme activity was markedly stimulated by calcium ion and completely inhibited by cobalt and mercuric ions and by ethylenediaminetetraacetate. Pectic acid or pectin with lower methoxyl content was a good substrate for this enzyme, while no significant activity was observed when pectin with higher methoxyl content was used as a substrate. It was concluded that the enzyme produced under the normal growth conditions is an endo-pectate lyase and differs from the pectin lyase induced by nalidixic acid.

View full abstract

The specific surface area and the specific pore volume distribution of Japanese tobaccos were measured by means of the low temperature gas adsorption technique utilizing the B. E. T. and the Inkley methods. The specific surface area and the specific pore volume of the micropores less than 300 Å in diameter varied from 6, 000 to 17, 000cm²/g and from 0.0012 to 0.0036cm³/g, respectively, with types of curing in the ascending order of the sun cured, the flue cured and the air cured tobaccos. The both specific values were increased by extracting the tobaccos with water greatly in the case of the flue cured, while slightly in the case of the air cured tobaccos, suggesting that the effect of the curings on the specific values were due to differences in the content of low molecular components. Effectiveness of puffing was also shown. The specific surface area was linearly correlated with the specific volume of the micropores less than 300 Å in diameter, the constant term showing that contribution of the larger pores more than 300 Å in diameter to the specific surface area of tobacco was insignificant.

View full abstract

About 1, 200 microbial cultured filtrates have been screened for inhibition of cyclic nucleotide phosphodiesterases (EC 3. 1. 4. c] prepared from rabbit brains. One of the active agents was isolated and identified as reticulol (6, 8-dihydroxy-7-methoxy-3-metbylisocoumarin).

View full abstract

It was found in the previous studies that the quality of flour is closely related to the aggregation behavior of gluten separated from it, and that difference of aggregation behavior of gluten was mainly due to the nature of glutenin contained in it. In the present paper, aggregation behaviors of glutenins were investigated in relation to their polypeptide compositions with ten kinds of flours. Component polypeptides of glutenin were fractionated into three fractions, FI, FII and FIII, by gel filtration, and their polypeptide compositions and aggregation behaviors were investigated. The ratio of the three fractions was different among the ten glutenins, and a correlation was found between τ₁₀/C values, a parameter for aggregation reaction, of glutenins and their FI and FII contents. τ₁₀/C decreased with increasing content of FI and with decreasing content of FII. Therefore, the aggregation behavior of gluten depends at least partly on the contents of FI and FII in glutenin.

View full abstract

Since amino acids and saccharides have been used as model substrates for immobilized enzymes, the diffusion coefficients of those compounds were measured in the solutions of glucose polymers, such as dextran, DEAE-dextran and dextran sulfate which are the constituents of carriers of immobilized enzymes, by the height area method using the Schlieren optical system. The diffusion coefficients decreased with an increase of the polymer concentration. The decrease of diffusion coefficient was most significant in L-arginine-dextran sulfate system where the former had a positive net charge, while the latter the negative ones. A model by which the experimental results could be explained was proposed. In the model, were taken into account the direct interactions between the diffusing solute and the polymer, i.e., the hydrogen bonding and the ionic interaction. A fairly good agreement was observed between the experimental and calculated results.

View full abstract

A beer-gushing inducing protein (NGF) produced by Nigrospora sp. No. 207 was found to be characterized by a relatively higher content of hydrophobic amino acid residues, a considerably high content (about 10%) of half-cystine residue, and the lack of tyrosine, tryptophan, methionine and histidine. Cleavage of disulfide bonds in NGF, either oxidative or reductive, resulted in complete loss of the gushing-inducing activity, indicating that the disulfide bonds are essential for exhibiting the activity.
Modification of amino groups in NGF with maleic anhydride or O-methylisourea did not affect the gushing activity, whereas modification with trinitrobenzene sulfonate reduced the activity to some extent. Modification of arginine-guanidino groups in NGF with phenylglyoxal markedly reduced the gushing activity. When the free carboxyl groups of NGF were modified by incorporating glycine or tyrosine, the gushing activity was also drastically destroyed, implying that the acidic groups of the NGF protein are important for the gushing activity.
It appeared that maintenance of a definite molecular conformation is essential for the gushing-inducing activity of this hydrophobic protein.

View full abstract

The acidic constituents of sun-cured Turkish tobacco have been studied, Of the 93 acidic compounds investigated, 10 compounds are new in nature and 18 are new tobacco constituents. The 93 compounds fall mainly into three groups: fatty acids, aromatic acids, and terpenoid acids, which appear to be derived from macrocyclic thunbergane diterpenoids.

View full abstract

Seventeen bacterial capable of utilizing phthalate esters isolated from natural sources were identified. Based on morphological and biochemical characteristics, type of cell division, GC content in DNA, principal amino acid in the cell wall and cellular fatty acid composition, 10 isolates were identified as Nocardia erythropolis, one isolate as Pseudomonas acidovorans, another as Pseudomonas cepacia and four as members of the genus of Corynebacterium.

View full abstract

A comparative study of inhibition on the substrate-incorporation in several species of tumor cells has been achieved in combination with the antibiotics having different action mechanisms. It was thus revealed that a large part of the antitumor antibiotics so far examined showed a marked inhibitory response even at a low concentration. Particularly, the antibiotics whose action mechanism were established primarily on cell membrane were markedly sensitive. An apparent difference in the sensitivity was observed in some groups of antitumor antibiotics which act on nucleic acid synthesis. A methodological survey together with an evaluation of this procedure is discussed.

View full abstract

A new antitumor substance designated renastacarcin has been isolated from the cultured broth of Streptomyces violascens by means of DEAE-cellulose and Sephadex G-100 column chromatography. The homogeneity of renastacarcin was confirmed by electrophoreses on a polyacrylamide gel and a cellulose acetate membrane as well as by ultracentrifugal analysis. A sedimentation coefficient at 55, 430 rpm was found to be 2.12 S and the molecular weight was calculated to be 34, 500.

View full abstract

The purified sample of renastacarcin was tested against various kinds of Gram-positive and -negative bacteria, yeast and fungi. As a result, it was concluded that this compounds was almost devoid of antimicrobial activity even at a concentration of 200 mcg/ml. However, renastacarcin showed a significant antitumor activity both in vitro and in vivo against ascitic type of Ehrlich carcinoma and Sarcoma 180 in mice. When the tumor cells were incubated with renastacarcin, the lysis of cell membrane and subsequent leakage of the cellular contents were observed by means of the optical and microscopical method.
The LD₅₀ values of renastacarcin in mice were observed to be 40mg/kg intraperitoneally, and 23.5mg/kg intervenously.

View full abstract

One of the bound forms of vitamin B₆ occurring in rice bran was isolated in a faintly yellowish syrup by repeating ion-exchange and paper-partition chromatographic techniques. The behaviors of the isolate on thin-layer and Aminex A-5 column chromatograms were coincident with those of synthetic pyridoxine-β-D-glucoside which was obtained by Königs-Knorr condensation of α⁴, 3-O-isopropylidene pyridoxine and 2, 3, 4, 6-tetra-O-acetyl-α-D-glucopyranosyl bromide. On acid hydrolysis, the isolate gave pyridoxine and glucose. Glucose was proved to bind to the 5-hydroxymethyl group of pyridoxine, because the isolate did not react with 2, 6-dichloroquinone chlorimide in the presence of boric acid. An equimolar amount of pyridoxine and D-glucose was produced with an equivalent consumption of the isolate by the action of β-glucosidase. No essential difference between the isolated and synthetic preparations could be detected in UV- and NMR-spectral features. Thus, the chemical structure of the isolate was identified as 5'-O-(β-D-glucopyranosyl) pyridoxine.

View full abstract

Hemagglutinin was extracted from castor beans and purified by gel filtration on Sephadex G-75 followed by DEAE-cellulose column chromatography. The molecular weight of castor bean hemagglutinin (CBH) was estimated to be 130, 000 by SDS-polyacrylamide gel electrophoresis and its isoelectric point to be 7.8 by isoelectric focusing electrophoresis. CBH possessed the identical N-terminal amino acids (Ile and Ala) and a very similar amino acid composition to those of ricin D, whereas its toxicity for mice was about 1% of that of ricin D.

View full abstract

Radioactive gibberellin A₂₀ (GA₂₀, I) was prepared by hydrogenation of GA₅ methyl ester-16, 17-epoxide with tritium gas followed by removal of epoxide oxygen and hydrolysis. Radioactive GA₅(V) was obtained by hydrogenation of GA₃ methyl ester monomesylate with tritium gas followed by hydrolysis and radioactive GA₈ (XV) was obtained by oxidation of radioactive GA₅ with osmium tetroxide.

View full abstract