NADPH-cytochrome c oxidoreductase obtained from the cells of Thiobacillus thiooxidans displayed the pH optimum of 8.7 and was completely inactivated by heating at 60°C for 10 min. Enzymatic activity was proportional to protein concentration and linear with time. The Km for NADPH was found to be 2.13×10^-5M. The enzyme was specific for NADPH and transferred electrons to cytochrome c and some dyes. p-Chloromercuribenzoate, EDTA and acryflavin inhibited the reductase activity.

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Volatile smelling compounds of freezed egg white, freshness of which was kept by freezed storage, were collected by steam-distillation. After DNP-hydrazones of volatile carbonyl compounds were separated into four classes by column chromatography, DNP-hydrazones contained in each class were separated by thin layer chromatography. Rf and melting point of recrystalized compounds were compared with those of authentic compounds. Volatile basic compounds were collected as hydrochlorides and detected by paper and thin layer chromatography.
Acetone, acetaldehyde, formaldehyde, 2-pentanone, 2-butanone diacetyl except two unknown compounds as volatile carbonyl compounds, and ammonia, methylamine, dime-thylamine and putrescine as volatile basic compounds were tentatively identified.
Correlations between these compounds and smell of freezed egg white were discussed.

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A modification of the cysteine-sulfuric acid method of Dische and Devil¹¹⁾ is described. The modified procedure is based on the observation that fructose and glucose give different time course of color development. By this modified procedure, fructose in the presence of 100-fold excess of glucose can be determined with an error of about 10%.

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Modifications of the anthrone-sulfuric acid method and the phenol-sulfuric acid method are described. By employing the principle of two-point determination of Mokrasch and by modifying the conditions for color development, fructose in the presence of 100-fold excess of glucose can be determined with an error of about 15% by the modified anthrone-sulfuric acid method. The modified phenol-sulfuric acid method also gave the same order of sensitivity and specificity to fructose as the modified anthrone-sulfuric acid method.

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A number of bacterial strains from type culture collections and natural sources were examined in their metabolic characteristics toward sorbitol and L-sorbose.
Paper chromatographic analyses of sorbitol and L-sorbose metabolites obtained from the cultures of various bacteria revealed that the organisms producing 2-keto-L-gulonic acid from sorbitol were merely found in the genera Acetobacter, Gluconobacter and Pseudomonas, whereas those producing the acid from L-sorbose were distributed in the twelve genera of bacteria: Acetobacter, Alcaligenes, Aerobacter, Azotobacter, Bacillus, Escherichia, Gluconobacter, Klebsiella, Micrococcus, Pseudomonas, Serratia and Xanthomonas. G. melanogenus, which was characterized by excellent production of 2- keto-L-gu Ionic acid from sorbitol, also formed several other sugars and sugar acids as the sorbitol metabolites. These compounds were identified to be D-fructose, L-sorbose, D-mannonic acid, L-idonic acid, 2-keto-D-gluconic acid and 5-keto-D-mannonic acid, respectively, by means of two-dimensional paper chromatography.
Bacteria producing 2-keto-L-gulonic acid from sorbitol were usually isolated from fruits but not from soil.

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The location of (1→3)-D-glucosidic linkages in a dextran elaborated by Leuconostoc sp. IFO 12370 has been investigated by the Smith degradation technique. The polyaldehyde produced by periodate oxidation of the dextran was reduced with borohydride to the corresponding polyalcohol. Complete acid hydrolysis of the dextran polyalcohol yielded glycerol and glucose, in the molar ratio of 17:1. The polyalcohol gave upon mild acid hydrolysis glycerol and a small proportion of α-1-O-D-glucosylglycerol, which arose from the sequence →6G1→3G1→6G1→ in the parent dextran. Methylation of the polyalcohol followed by acid hydrolysis gave 1-O-methyl-glycerol (17 parts) and 2, 4-di-O-methyl-D-glu-cose (1 part), together with a trace of 2, 4, 6-tri-O-methyl-D-glucose. These results indicate that most of (1→3)-linkages in the dextran molecule are located as the branch points of the (1→6)-linked chain. The lengths of repeating unit in the various fractions of acid-degraded dextran, estimated from the ratio of glycerol and glucose derived from the poly-alcohols, were not significantly different from that of the native dextran, suggesting that random cleavage of the chain occurs during partial acid-hydrolysis.

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A method has been described for the isolation and purification of two basic hemo-proteins, cytochrome c and peroxidase 556, from 70kg of rice embryo. The hemoproteins were extracted with dilute phosphate solution without any pretreatment with organic solvent. Ion exchange chromatography on CM-Sephadex C-50 and gel filtration chromato-graphy on Sephadex G-75 and G-100 were successfully applied for the purification. Both hemoproteins as major constituents were crystallized from ammonium sulfate solution, and some other chromoproteins, a blue protein and a flavoprotein, were also purified. Crystalline cytochrome c of rice embryo was found to be quite similar to cytochrome c of mammalian sources. Peroxidase 556 was found to spectroscopically belong to high spin form of peroxidases such as horseradish peroxidase II and Japanese-radish peroxidase a but it was a basic protein unlike to the latter two. Physiological significance of these hemoproteins is briefly discussed.

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This paper constitutes the second in a research series undertaken to clarify the inter-active relationships among soybean meal protein, calcium and phytic acid. The extent of bound calcium by cold insoluble fraction protein which is low in phosphorus content was examined by differential equilibrium dialysis, differing concentrations of calcium and phosphorus. The results indicate clearly that many molecules of calcium and phosphorus may be bound by a single protein molecule and that extent of calcium increases as phosphorous compounds in dialysis system such as phytic acid increase. The maximum number of bound calcium was calculated after statical equation by Klotz.

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Volatiles stripped from rice bran are analyzed by gas chromatography and thin-layer chromatography either directly or after converted into 2, 4-DNPH's or 3, 5-DNB's. Most of prominent peaks revealed by a direct gas chromatography of the volatiles condensates behave as alcohols against classification reagents and are identified as methanol, ethanol, n-propanol, sec-butanol, isobutanol, n-butanol, isopentanol, n-pentanol and n-hexanol, re-spectively. Ethanol and methanol, being major components in the condensates are also detected by thin-layer chromatography of their 3, 5-DNB's. Carbonyls identified by a flash exchange gas chromatography of 2, 4-DNPH's of the volatiles are ethanal, propanal, isobutanal, acetone, n-butanal, isopentanal, n-pentanal and n-hexanal. Thin-layer chromato-graphy of 2, 4-DNPH's reveals presence of ethanal, acetone, isopentanal or/and isobutanal and n-hexanal in the volatiles. The experimental data have been integrated to furnish a basis for extensive comparison with other rice products and by-products.

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HM 2 phage (group I) of Clostridium saccharoperbutylacetonicum was studied with regard to its peculiar behavior on exposure to deoxyribonuclease (DNase). It ss as inactivated by DNase: 10, 50 and 100μg/ml of DNase inactivated 20, 50 and 70% of free HM 2 phages, respectively. Whereas, JIM 3 phage (group II) was not affected by even 300μg/ml of DNase.
The mechanism of the inactivation of HM 2 phage by DNase was supposed either to be the enzymatic digestion of phage DNA by DNase, or to be a polyelectrolytic interaction between the phage protein and DNase. On these subjects, the following results were obtained. (1) Heat-treated DNase, the enzymatic activity of which was destroyed, did not affect on HM 2 phage. (2) HM 2 phage was inactivated by basic proteins which caused polyelectrolytic interaction with phage. Whereas, HM 3 phage, which was not inactivated by DNase, was inactivated more intensively than the former by basic proteins. (3) HM 2 phages in high concentration formed large aggregates with basic proteins, but not with DNase. (4) When EM 2 phages were exposed to DNase, the large amount of phage-DNA was released from phage coat membranes. These DNA were considered to be digested by DNase in phage particles.
These results indicated that the inactivation of HM 2 phage by DNase was due to the enzymatic action of DNase.

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Strejtomyces kanamyceticus produces ¹⁴C-kanamycin and other labeled kanamvcin-related compounds in the presence of ¹⁴C-glucose or ¹⁴C-glucosamine. One of the latter is paromamine. In order to find a condition to obtain high incorporation rate of radioactive precursors, the culture phase for the addition of labeled compounds was studied. The maximal incorporation rate was obtained by addition of ¹⁴C-glucosamine in the later phase of the culture and the specific activity of resultant kanamycin A was found to be much higher than in the case of the addition of ¹⁴C-glucose. In some experiments one third of radioactivity of the added ¹⁴C-glucosamine was taken into kanamycin, and the distribution was limited only in 6-amino-6-deoxy-D-glucose moiety of kanamycin A when ¹⁴C-glucosamine was added. In contrast, when ¹⁴C-glucose was added, approximately equal distribution of radioactivity was observed in each of three moieties of kanamycin A molecule.

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The usefulness of partially hydrolyzed polyvinyl alcohol (PVA_??_=500) was examined by the absorption and excretion from the gastro-intestinal tract of adult rat. About 86% of orally injected PVA was excreted in feces in 48 hr, but no excretion was observed in urine. More than 50% of PVA injected into heart was excreted in urine in 12hr, but no fecal PVA was observed. About 100%, of orally injected PVA was recovered in the gastro-intestinal tract killed 5.5hr after injection, but no PVA was observed in blood from the killed rats.

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A characteristic effect of inorganic neutral salts on myrosinase was discovered. The salts containing monovalent anions had a remarkable inhibitory effect on the ascorbate-activated enzyme, but little on the non-activated enzyme. Such an effect was elucidated to be due to the anion. For the ascorbate-activated enzyme, a linear relation was obtained by plotting the logarithms of the enzymatic activity against the square roots of the ionic strength of the salts. Therefore, the effect of monovalent anions is ascribable to the ionic strength of the solvent. The Km values of the activated and non-activated enzyme were increased by the presence of the monovalent anion.

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Three different types of proteinase inhibitors, I, II and III, were fractionated from Japanese radish seed by repeated column chromatographies on SE- and CM-cellulose. The finally purified preparation of inhibitor III was found to be homogeneous by both chroma-tographic and electrophoretic analyses. All three of them strongly and stoichiometrically inhibited trypsin whereas they showed weak inhibition on other proteinases, such as chymotrypsin, Nagarse and Pronase. From nitrogen content and ultraviolet absorption spectra, each of the inhibitors I and III .vas confirmed to be a protein. The molecular weights of inhibitors I and III were calculated to be 8000 and 12, 000, respectively. These inhibitors were stable at temperatures above 90°C in an acidic pH.

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(±)Sclerin (4, 5, 6, 7-tetramethyl-8-hydroxyisochroman-1, 3-dione), a mycerial extracts of Sclerotinia libertiana, and related compounds were synthesized. Alkylated-7-hydroxyindanones were obtained from alkylated phenols and α-bromobutyrylbromide, and these indanone derivatives were treated with benzaldehyde to give 2-benzylidene-7-hydroxyindanone deri-vatives. By ozonolysis of these benzylidene-indanones were obtained 8-hydroxyisochroman-1, 3-diones.

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In order to obtain information of the gross-structure of glutelin, chemical and physico-chemical properties of S-cyanoethyl glutelin were investigated. Glutelin remained at the origin in polyacrylamide gel electrophoresis, while S-cyanoethyl glutelin migrated in the gel and resolved into two components. The ion-exchange chromatography by carboxymethyl Sephadex C-50 gave further resolution of S-cyanoethyl glutelin into one neutral component corresponding to the anodic component and two basic components corresponding to the cathodic component in polyacrylamide gel electrophoresis at neutral pH. The amino-terminal residue of the neutral component (Component I) could not be detected by the fluorodinitrobenzene method, while both the basic components (Component II and III) had only glycine as the amino-termini. On the basis of dinitrophenyl-glycine found, the minimum molecular weights of Component II and III were calculated at about 35, 000 and 43, 000 respectively. The relative concentration of these three components was as follows; Component I: Component II: Component III=8:1:1. These facts obviously indicate that glutelin is a very large molecule composed from these three components polymerized by disulfide linkage, Component I being the major subunit.

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From the volatiles of roasted barley or those formed during roasting of barley, furfural, 2-methylbutanal, 2-methylpropanal, 3-methylbutanal, 2, 3-pentanedione, ethylglyoxal and pyruvaldehyde were isolated and identified as their mono- or bis-2, 4-dinitrophenylhydrazones.
Removal of the carbonyl compounds from the volatiles resulted in a loss of the characteristic aroma of roasted barley.

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Some physicochemical properties of the acid-stable α-amylase and the acid-unstable a-amylase, produced by Asp. niger simultaneously, were investigated comparing with each other. The molecular weights of the acid-stable α-amylase and the acid-unstable α-amylase were computed from sedimentation equilibrium as 58, 000 and 61, 000, respectively. And the molecular shapes were investigated by measuring sedimentation coefficient and intrinsic viscosity. The results indicated that these two α-amylases were similar in hydrodynamic properties and that they were prolate ellipsoids and poor hydrated proteins. But high frictional ratio of the acid-stable α-amylase suggested a certain degree of molecular asymmetry. Isoelectric points of the acid-stable α-amylase and the acid-unstable α-amylase were deter-mined as pH 3.44 and 3.70, respectively.

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