The application of the radiopasteurization method to fermentation media consisting mainly of molasses was investigated. γ-Irradiation was found to have an excellent pasteurization effect on the fermentation media and at the same time to bring about an increase in the fermentation rate and yield of ethanol. Percent survivals in molasses decreased to ca. 70% by heating at 80°C for 30min, to ca. 10% by irradiation with 3.0×10⁵ rad and to ca. 1% by 6.0×10⁵ rad. Irradiated mash was suitable for the medium of the “starter”, since the rate and the degree of the growth of Saccharomyces cerevisiae in irradiated mash did not differ from those of the growth in heat-pasteurized mash.
In the case of the molasses mash supplemented with nitrogen sources, the fermentation rate and yield of ethanol in irradiated mash were larger than those in heated mash. Besides, in the absence of nitrogen sources a 14% difference in fermentation yield was seen between the mash irradiated with 3.0×10⁵ rad and the mash heated at 80°C. With the doses ranging from 1.0×10⁵ to 9.5×10₅ rad, concentrations of total sugar and direct reducing sugar, pH, and optical density of molasses were little affected by irradiation.

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1. Both amounts of lipid phosphorus and acid-insoluble nitrogen in the mitochondrial fraction from chilling-injured sweet potatoes (var. Okinawa 100) were larger than in the fraction from healthy sweet potatoes. The N-amount appeared to be increased more by chilling-injury than the P-amount.
2. Sweet potato, a tropical plant, showed lower value of the degree of unsaturation of fatty acids in mitochondrial fraction than white potato, a temperate-zone plant.
3. The amount of unsaturated fatty acids of C₁₆, C₁₈ and C₂₀ as percentage of the total fatty acids was higher in mitochondrial fractions from chilling-injured sweet potatoes (var. Okinawa 100 and var. Norin 1) than in mitochondrial fractions from healthy sweet potatoes. However, in the case of white potato mitochondrial fraction no detectable difference was observed between storage at 0_??_1°C and at 10_??_14°C.

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The inactivating action of monovalent cation (Na⁺) and the stabilizing action of divalent cation (Mg²⁺) on phage were studied using HM 2 phage (group I) of Clostridium saccharoperbutylacetonicum.
The maximum rate of the phage inactivation occured at 0.1M Na⁺. However, the serum-brocking power (SBP) of phage protein decreased with increasing the concentration of Na⁺. The inactivation of the phage by Na⁺ was associated with alteration of its sedimentation rate (s_{20, w}, ), the release of its DNA, and denaturation of its protein. These facts indicate that Na⁺ causes the destructions of the structures of phage protein and DNA-protein complex. The action of Na⁺ on the phage could be illustrated by the salting-in and salting-out effects of protein of the phage by Na⁺.

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Effects of purine and pyrimidine analogues on growth and amino acid accumulation were studied using Aerobacter aerogenes No. 19-35 which produces L-valine in phosphaterich medium (PS-medium) and L-glutamic acid in phosphate-poor medium (PD-medium). The cell growth was inhibited in PS-medium containing purine and pyrimidine analogues. In PD-medium, 6-thiolpurine such as 6-mercaptopurine, 6-methylmercaptopurine and 6-thioguanine did not inhibit cell growth while other drugs did. The addition of 6-thiolpurine, especially 6-mercaptopurine, to PD-medium provoked the production of valine in place of L-glutamic acid. Cells are classified into two types by these characters: L-valine producing type and L-glutamic acid producing type. Such conversion phenomena seemed to occur only in strain No. 19-35, since other strains of A. aerogenes and E. coli producing valine did not show the conversion phenomenon when exposed to 6-thiolpurines.

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From the first crop of P. graveolens and the first and third ones of P. roseum Bourbon, low-boiling compounds escaping during the steam distillation were collected, and examined through gas chromatography and the preparation of their derivatives. Nine hydrocarbons, five carbonyl compounds, three alcohols, two esters and one sulfide were identified. Among these twenty compounds, sixteen compounds except dimethyl sulfide, α-pinene, myrcene and limonene were newly identified in the geranium species. Appreciable variation between the species was observed in contents of methanol, acetone, dimethyl sulfide and terpene hydrocarbons.

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A mold which produced α-galactosidase and little invertase was isolated and identified as Mortierella vinacea. α-Galactosidase formation of the mold was induced by galactose, melibiose, raffinose and lactose. Among these inducers lactose showed the most stimulative effect. α-Galactosidase was produced by either Koji method or submerged culture method, but in the latter most α-galactosidase was found in the mycelium fraction.
Hydrolysis of raffinose in beet molasses was studied with the α-galactosidase in the mycelium fraction and about 80% of raffinose was found to be hydrolyzed by the enzyme preparation.

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In attempts to obtain GMP producing strains, Brevibacterium ammoniagenes was treated with UV, N. T. G. or D. E. S. as a mutagen. Adenine-guanine requiring mutants were obtained from an adenine-requiring mutant of Brev. ammoniagenes, KY 3482-9 and two of them, presumably adenine-xanthine requiring mutants, were then reverted to mutants which required only adenine for their growth.
Although these revertants were not able to accumulate a copious amount of GMP, most of them and of adenine-guanine requiring mutants produced larger amounts of IMP than the parent adenine-requiring strain.
Effects of Mn²⁺ and purine bases in the medium on IMP production by these mutants were examined and IMP productivities of these mutants were compared with the parent strain under optimal conditions.
These mutagenic treatments were thus proved to be effective for the increase of de novo IMP production by Brev. ammoniagenes mutants.

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Brevibacterium ammoniagenes ATCC 6872 accumulates 5'-GDP and -GTP, or 5'-ADP and -ATP together with GMP or AMP in nucleotide fermentation by salvage synthesis.
With cell free extract of this strain, transphosphorylating reactions of AMP or GMP were investigated.
ATP-AMP transphosphorylating enzyme (s) was partially purified to 21.7 fold with acid treatment, salting-out and column chromatography.
In ATP-AMP and ATP-GMP transphosphorylating reactins, optimal conditions were decided such as for concentrations of enzyme, of MgCl₂ and of phosphate donor, pH and cell age as the enzyme sources.
Specificities of phosphate donors and acceptors were examined with both the partially purified enzymes or the sonicate. AMP and GMP were phosphorylated by ATP rapidly, but IMP and XMP were not, therefore supporting our previous finding that Brev. ammoniagenes could not accumulated IDP, ITP, XDP and XTP in IMP and XMP fermentation, respectively.
Although ATP was the best donor for both AMP and GMP phosphorylations, other nucleoside triphosphates and PRPP were used as phosphate donors.
Furthermore, phosphorylation of ADP to ATP was investigated and possible mechanisms of nucleoside di- or triphosphates synthesis in the nucleotide fermentation were discussed.
From these results, it is suggested as a possible mechanism for nucleoside di- and triphosphate accumulation by Brev. ammoniagenes, that a nucleoside monophosphate formed is phosphorylated to a nucleoside di-phosphate with ATP or other phosphate donors and then the nucleoside diphosphate is converted to a triphosphate with these phosphate donors.

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Both AMP and GMP were transphosphorylated rapidly to the corresponding nucleoside-diphosphates and triphosphates by ATP and by other high energy phosphate compounds with cell free extracts of Brevibacterium ammoniagenes.
Some enzyme inhibitors, such as metals and PCMB were shown to inhibit the phosphorylations of AMP and GMP. Higher levels of ATP, ADP, GTP and GDP also inhibited the activity of the partially purified ATP-AMP transphosphorylating enzyme (s).
In guanine nucleotides fermentation by salvage synthesis with this strain, addition of these inhibitors to the medium increased the amounts of GMP and total guanine nucleotides accumulated.
On the contrary, supplement of xylene or of other organic solvents to the medium stimulated the accumulation of bothe GTP and total guanine compouuds in this fermentation. From enzymatic studies, these solvents are presumed to have the ability to change cell permeability.
Such findings give an effective method for controlling the amounts of nucleotides accumulated in these fermentations.

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Changes in ribulose diphosphate (RuDP) carboxylase activity and the content of fraction 1 protein, which had been elucidated to be identical with protein of RuDP carboxylase, in tobacco leaves were examined with age, comparing with change in total protein content. Fraction 1 protein was determined by an immunological precipitin method developed in this experiment. Fraction 1 protein decreased with age and the rate was similar or a little larger than those of total protein and total chlorophyll. The carboxylase activity decreased more rapidly than fraction 1 protein during the senescent process. In a plant, upper leaves showed higher values of the carboxylase activity and fraction 1 protein content than lower leaves. The specific activity, RuDP carboxylase activity per unit fraction 1 protein, in upper leaves was higher than that in lower leaves.

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A protease inhibitor produced by Penicillium cyclopium on solid sultures of wheat bran was purified by means of column chromatography on Duolite A-2 and DEAE-cellulose. acetone precipitation and lyophilization. The purified inhibitor obtained as a white, floccose and hygroscopic substance was monodisperse by ultracentrifugal analysis. It was found to be an acidic macro-molecule of a molecular weight of about 5000. The chemical analyses rejected the possibility of the presence of amino acids, peptides, sugars, amino sugars, or uronic acids in the inhibitor molecule.

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Properties of a protease inhibitor from Penicillium cyclopium were studied. The pH range of the inhibitor action is restricted to acid pH, optimally at pH 3. Increasing temperature accelerates its action upon enzyme. The inhibitor causes enzyme inactivation in proportion to its concentration. It is fairly stable in an acid solution but unstable in an alkaline solution. It undergoes destruction by heat, hydrogen peroxide and ascorbic acid. The inhibitor reversibly combines with Al³⁺, Fe³⁺, Ag⁺ and Cu²⁺ to produce a precipitate. Salts interfer with the inhibitor activity. Generally, acid proteases from various penicillia are susceptible to the inhibitor while those from other genera are resistant.

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The behaviour of 3-deoxyglucosone (3-D-G) was investigated in the process of browning of Saké, and it was concluded that the browning of Saké involved at least three kinds of browning reactions which were the amino-carbonyl reaction, the caramelization and some other browning reaction by components other than glucose in Saké. The active dicarbonyl compound of 3-D-G was confirmed to act as an important intermediate in both the amino-carbonyl reaction and caramelization. The caramelization was the browning of glucose by itself via 3-D-G, while the amino-carbonyl reaction was the browning of glucose via 3-D-G by the interaction with the amino compounds in Saké. The remainder was, however, independent of glucose and 3-D-G. The temperature coefficient for the rate of the overall browning of Saké was estimated to be 2.8 to 2.9 of Q₁₀ value.

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In the previous report, it has been concluded that the browning of Saké involves three kinds of browning reactions which are the amino-carbonyl reaction, the caramelization and some other browning reaction by components other than glucose. In the present work, the extents of contributions of the three kinds of reactions to the browning of Saké were actually estimated, respectively, by analysing its dependence on the glucose concentration. It was concluded that the browning by components other than glucose was the major factor while the amino-carbonyl reaction and the caramelization were the minor ones in the browning of Saké. In addition, the change in pH value, as well as in temperature, was observed to influence exponentially on the browning rate of Saké. An empirical equation was proposed to elucidate the way in which the three kinds of reactions con-tributed to the overall browning in Saké.

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The amino acid compositions of the acid-stable α-amylase and the acid-unstable α-amylase obtained from Aspergillus niger were determined by automatic column chromatography. The amino acid composition of the acid-unstable α-amylase was very similar to that of the a-amylase of Aspergillus oryzae. The amino acid composition of the acid-stable α-amylase was also similar in most part, but differed from that of the acid-unstable α-amylase in the following features. (a) The lysine content was lower. (b) Although the totals of carboxyl and amide were almost equal, there were considerably more free carboxyl residues. (c) The serine content was higher. (d) The proline content was lower. These facts may be related to the lower isoelectric point (pH 3.44) of the acid-stable α-amylase.
Amino-terminal amino acid analysis demonstrated one mole of amino-terminal leucine or isoleucine per mole of the acid-stable α-amylase and one mole of amino-terminal alanine per mole of the acid-unstable α-amylase.

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Some physicochemical properties of the cathepsin D purified from the rabbit muscle (L. dorsi) were investigated.
The sedimentation coefficient (S_{20, w}) and the molecular weight determined from sedimentation equilibrium experiment was 3.83S and 29, 000_??_30, 000, respectively.
The amino acid composition of the enzyme was determined with an automatic amino acid analyzer.
The proteolytic specificity of the enzyme was also investigated using the B-chain of oxidized beef insulin as the substrate. The cathepsin D cleaved the bonds Phe-Val, Ala-Leu, Leu-Tyr and Tyr-Leu. The specificity of the cathepsin D was fairly similar to that of the pepsin.

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1. It was demonstrated by silica gel thin layer chromatography that leucine-U-¹⁴C was incorporated into furanoterpenes, e. g. ipomeamarone, in sweet potato root tissue infected by Ceratocystis fimbriata.
2. Further proof for ipomeamarone synthesis from leucine-U-¹⁴C was obtained by the constancy of the specific radioactivity of ipomeamarone semicarbazone through repetitive crystallization.
3. The synthetic pathway of ipomeamarone from leucine was found to be connected with the synthetic pathway from acetate at least at some steps.
4. Leucine-U-¹⁴C was incorporated into both saponifiable and non-saponifiable materials in the same way as acetate-2-¹⁴C.

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Two novel gibberellins, GA₂₁ (I) and GA₂₂ (II), were isolated from immature seeds of sword bean, Canavalia gladiata DC. The isolation procedure of these substances as well as their growth-promoting effects on dwarf maize mutants d₁ and d₅, rice, cucumber and dwarf peas (Progress No. 9) are described.

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The structures of two new gibberellins, GA₂₁ and GA₂₂, isolated from immature seeds of sword bean, were determined as 4aα, 7α-dihydroxy-8-methylenegibbane-1, 1, 10β-tricar-boxylic acid 1→4a lactone (II) and 4aα, 7α-dihydroxy-1β-hydroxymethyl-8-methylenegibb-2-ene-1α, 10β-dicaboxylic acid 1→4a lactone (IV), respectively, on the basis of chemical and physicochemical studies.

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Justicidin A, C₂₂H₁₈O₇, mp 263°C and B, C₂₁H₁₆O₆, mp 240°C were isolated as fish-killing components from Justicia Hayatai var. decumbens. The piscicidal activities of both compounds were demonstrated to be as strong as rotenone and about ten times stronger than that of pentachlorophenol.

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Attempts were devoted to use Streptomyces aureofaciens and Streptomyces erythreus, the antibiotics producers as sources for the biosynthesis of cobalamine. The constituents of the fermentation medium and the strain play an important role in the biosynthesis of vitamin B₁₂. The same strain produced different amounts of antibiotic and vitamin on the two different constitutive media. The increase of the phosphorus concentration in the fermentation medium-within limits-increased the vitamin B₁₂. The optimal concentration of phosphorus favourable for the synthesis of cobalamine was inhibitive for the antibiotic production. The phosphorus level in the fermentation medium plays an important role in the metabolism of carbohydrate and consequently on the biosynthesis of antibiotics. Low concentration of 5, 6-dimethylbenzimidazole (cobalamine precursor) in the presence of suitable phosphorus induced the microorganism to increase its biosynthetic potentiality for the vitamin B₁₂ production.

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