Shikimic acid-G-¹⁴C, phenylalanine-U-¹⁴C, and trans-cinnamic acid-3-¹⁴C were incorporated into the gallic acid moiety of ( - )-epicatechin-3-gallate and ( - )-epigallocatechin-3-gallate in young tea shoots. The incorporation of shikimic acid-7-¹⁴C into the gallic acid was almost equal to that of shikimic acid-G-¹⁴C, i.e. decarboxylation of the side chain did not occur during the conversion of shikimic acid to gallic acid and the conversion apparently was due to dehydrogenation of the shikimic acid. These results suggest at least two pathways for gallic acid biosynthesis in tea shoots;a) a pathway through phenylpropanoid, and b) a pathway through the dehydrogenation of shikimic acid.
Esterification of gallic acid with catechins was also confirmed by tracer experiments in young Tea shoots.

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Gamma-radiation effects on Taiwan-produced rice-grains have been investigated and discussed to find an advantageous application of radiation processing on the preservation of rice-grains.This paper deals with the results obtained from the three major kinds of rice-grains which were gamma-irradiated and stored.
The factors of contaminated microorganisms on the rice-grains, chemical analysis of rice-ingredients, activity of α-amylase in the rice-grains, germination-rate of the rice-grains, rice-insect fertility and cooking quality of the hulled rice were studied. Among them, the following factors were considered as the main results.
1. Gamma-irradiation could reduce the contaminated microorganisms remarkably as the gamma-dose was raised over 100 krad, e. g., reduction of the survivals of microorganisms to 19.62-39.76% with 100 krad, to 5.41 -9.09% with 300 krad, to 0.58-0.81% with 1 Mrad during the twelve months storage period.
2. Some fluctuations on the fat acidity and the contents of reducing sugars and thiamine in the irradiated rice-grains were observed in the period of twelve months preservation at 20-30°C.
3. The germination of rice-grains was reduced considerably with the irradiated samples exposed to a gamma-dose over 50 krad.
4. No deleterious effects on rice-insect fertility on the irradiated rice-grains with a 50 krad gamma-dose occurred during the twenty-four months storage period, unlike those of the unirradiated ones and the irradiated ones with gamma-doses below 30 krad.
5. No significant changes were found on the cooking quality of the hulled rices irradiated
below a gamma-dose of 300 krad and stored for twenty-four months at 20-30°C.

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A β-cyano-L-alanine (β-CNAla)-degrading enzyme in Pseudomonas sp. 13 was purified and isolated as a crystalline preparation. The purification procedure involved ethanol fractionation, ammonium sulfate fractionation, column chromatography on DEAE-cellulose and crystallization in the presence of ammonium sulfate, resulting in 660-fold purification with a yield of 21%.
The crystalline enzyme was homogeneous as judged by ultracentrifugation and SDS-polyacrylamide gel electrophoresis. The enzyme has a molecular weight of approximately 1, 000, 000, and consists of about 30 apparently identical subunits, each of a molecular weight of approximately 35, 000.
The enzyme catalyzed the hydrolysis of β-CNAla to form asparagine and aspartic acid. The enzyme reaction was inhibited strongly by thiol reagents.

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Volatile components of Zinchoge flowers were identified. Zinchoge flowers were extracted with pentane. The odor of the pentane extract was very similar to that of the Zinchoge flower. This extract was then fractionated into four fractions by distillation, and two of them were further separated by column chromatography into five fractions, respectively. These fractions were analyzed by gas chromatography and combined gas chromatography/mass spectroscopy.
One hundred and forty-five compounds, including 30 hydrocarbons, 39 esters, 20 aldehydes, 8 ketones, 21 alcohols, 7 phenols, 10 acids and 10 miscellaneous compounds, were identified. One hundred and nineteen of these compounds were newly identified as volatile components of Zinchoge flowers.

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The structure of an extracellular acidic polysaccharide (APS-I) of R. meliloti 201 was studied.Digestion of APS-I with APS-I depolymerase revealed that it consists of three repeating units of oligosaccharides, termed OP1, OP2 and OP3, in a ratio of 55:25:20 (by weight). ¹H-NMR analysis, methylation analysis, Smith degradation and α-D-mannosidase digestion indicated that OP1 was a pentasaccharide with the following structure:
β-D-Glcp
(1→4)
α-D-Manp-(1→4)-α-D-GlcUAp-(1→3)-α-D-Manp-(1→3)-D-Glcp
The structures of OP2 and OP3 were not fully elucidated, but each oligosaccharide had some structural similarities with OP1, containing a (1→4)-linked D-glucuronosyl residue, (1→3)(1→4)-linked D-mannosyl residue, (1→3)-linked reducing-terminal D-glucose residue and non-reducing-terminal D-mannosyl and D-glucosyl residues.

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A new glycopeptide antibiotic, OA-7653, was isolated and purified from the culture broth of an actinomycete, designated as Streptomyces hygroscopicus subsp. hiwasaensis subsp. nov. OA-7653 appears to be most analogous to vancomycin in that it is a water-soluble compound with an ultraviolet absorption maximum at around 280 nm, containing only glucose as a neutral carbohydrate component and active against Gram-positive bacteria. The antibiotic exhibited moderate antimicrobial activities against Gram-positive bacteria including Mycobacterium and showed very low toxicity. It inhibited specifically the synthesis of cell wall peptidoglycan in bacteria.

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Changes were investigated in trace alkaloid composition of MC-1 (bright) and Burley-21(burley) leaf lamina during curing. Seventeen alkaloids were identified. The total level of trace alkaloids decreased by approximately 60% in both MC-1 (flue-cured) and Burley-21 (air-cured).Although the level of most alkaloids also decreased, 2, 3' -bipyridyl and four nornicotine derivatives, i.e. N'-hexanoyl-, N'-octanoyl-, 1'-(6-hydroxyoctanoyl)- and 1'-(7-hydroxyoctanoyl)-nornicotine, increased during curing in both varieties. Differences in the trace alkaloid composition of cured leaf lamina among MC-1 (bright), Burley-21 (burley) and Basma (Orient) were also studied. In all varieties, N'-formylnornicotine, nicotyrine, anabasine and cotinine were present at high concentrations relative to the other trace alkaloids. The alkaloids which were present in a relatively high concentration in particular varieties, were as follows : MC-1 ; N'-formylanabasine, Burley-21 ; N'-hexanoyl- and N'-octanoyl-nornicotine, and Basma ; cotinine and N'-formylanatabine.

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An extracellular amylase produced by Bacillus circulans F-2 was purified to show a single band on SDS polyacrylamide gel electrophoresis. The purified enzyme has a molecular weight of 93, 000, an optimum pH of 6.0-6.5 and an optimum temperature of 60°C. On isoelectric focusing, the enzyme exhibited 2 bands with isoelectric points of 4.88 and 4.93. When acting on amylaceous polysaccharides, the enzyme removed maltohexaose from their non-reducing ends but the produced maltohexaose was then split into maltotetraose and maltose. Although it digested corn starch granules at approximately the same rate as porcine pancreatic and Streptococcus bovis amylases, it digested potato starch granules far faster than those amylases.

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Antioxidative activity was measured on extracts of rosemary (Rosmarinus officinalis L.), one of the herb spices belongs to the Family Labiatae, using different solvents. Strong activity was observed in the weakly acidic fraction of the n-hexane extract, which was further fractionated and purified to afford several active compounds. An ordorless and colorless compound, named rosmanol, showed high antioxidative activity in both lard and linoleic acid, and particularly in lard, was about four times more active than synthetic antioxidants, such as BHT and BHA.Furthermore, the antioxidant activities of the derivatives of rosmanol and carnosol were measured by the ferric thiocyanate method and the TBA method, to observe the correlation between chemical structure and activity as an antioxidant.

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Three serine proteinases designated as proteinases A, B and C were purified 800 to 2, 000-fold from an extract of Euphausia superba with an affinity column of soybean trypsin inhibitor (STI)-Sepharose 4B, and by gel filtration on Sephadex G-75 and DEAE-cellulose chromatography. The final preparations except for proteinase A were electrophoretically homogenous. The molecular weights of enzymes A, B and C were determined to be 28, 000-30, 000. Enzymes B and C showed very similar amino acid compositions in which acidic amino acids were abundant. Enzymes A, B and C could hydrolyze protein substrates and synthetic ester substrates including arginine residues. But they hydrolyzed most of the substrates more slowly than bovine trypsin did. All the enzymes were inhibited by soybean trypsin inhibitor, diisopropyl-fluorophosphate, leupeptin and antipain.Their enzymatic and molecular properties such as an instability in an acidic pH region, molecular weight and amino acid composition were similar to those of fish trypsin-like enzymes although krill enzymes had very low isoelectric points (pI 2.6).
These results suggest proteinases A, B and C in Antarctic krill are anionic trypsin-like enzymes and different from the well known cathionic trypsins in mammals.

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Wheat germ phosphatase (WP), sweet potato phosphatase (SP) and bovine spleen phosphoprotein phosphatase (BP) were immobilized on aminoalkylsilyl glass beads with glutaraldehyde, and denoted as IWP, ISP and IBP, respectively. Their activity yields were in the range of 16-44%. By this immobilization, the pH optima for WP and SP shifted one pH unit to the acid side and the temperature optima did not change. Their stabilities regarding pH and temperature were also comparable but their Km values were visibly increased compared with enzymes in the soluble state. The immobilized acid phosphatases were able to be used for limited hydrolysis of the phosphoryl group in casein. Rennet curd, with a smaller tension value, was obtained using test-milk dephosphorylated with IWP or ISP. The addition of CaCl₂ was found to slow down the phosphorylation of casein.

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Rice stored at 40°C and 80% relative humidity for 60 days, which was sensorially evaluated as old rice, showed a significant difference in cooking quality and texture of when cooked as compared with 4°C-stored rice. Free phenolic acids were detected in a larger amount in the 40°C-stored rice, and it is proposed that the increase of phenolic acids during storage may partly contribute to the cooking properties of old rice. GC analyses of the volatiles of cooked rice showed that a larger amount of pentanal, hexanal, heptanal, alkenals, ketones, 2-pentylfuran, 4-vinylphenol, etc., and a smaller amount of 1-pentanol, 1-hexanol, etc. were found in 40°C-stored rice than in 4°C-stored rice. The relationship between changes in these volatile components and the flavor of cooked old rice is also discussed.

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Two carboxyl proteinases with high milk clotting activity from Irpex lacteus designated as enzymes A and B were isolated by affinity chromatography with dehydroacetyl-pepstatin as ligand, DEAE-cellulose chromatography and isoelectric focusing. The final preparations were judged homogeneous by polyacrylamide gel electrophoresis. The molecular weights of enzymes A and B were determined by gel filtration and SDS-polyacrylamide gel electrophoresis to be 36, 000. The isoelectric points of A and B were 4.9 and 5.3, respectively.
A and B had similar amino acid compositions lacking sulfur-containing amino acids, such as cystine, cysteine and methionine. And both of the enzymes were inhibited by carboxyl proteinase inhibitors such as pepstatin, diazoacetyl-DL-norleucine methyl ester (DAN) and 1, 2-epoxy-(p-nitrophenoxy)propane (EPNP). These results indicate that enzymes A and B are similar to calf chymosin and other microbial rennets in their active site structure having two different carboxyl groups, although they showed minor differences in regard to enzymatic and molecular properties.Enzymes A and B exhibited almost the same ratio of milk-clotting activity to proteolytic activity as commercial microbial rennets obtained from Mucor pusillus and Mucor miehei.

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A differential pulse polarographic method based on the Brdicka current (polarographic catalytic hydrogen evolution current produced by proteins in the presence of the cobalt ion) was applied to the direct titration of subtilisin BPN' (S.BPN') with Streptomyces subtilisin inhibitor(SSI) at a concentration level of 10^-9 M. The first and second dissociation constants of the dimeric S.BPN'-SSI complex were determined by fitting theoretical curves to the titration data, in which the multiple equilibrium involving microscopically distinct forms of the S.BPN'-SSI complex was taken into account. The intrinsic free energy change in the first binding of S.BPN' to dimeric SSI was larger than that of the second binding.

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Shoot-forming cultures of Digitalis purpurea L. were grown in various modifications of Murashige-Skoog medium. The effects of CaCl₂, MgSO₄, FeSO₄ coupled with Na₂-EDTA, minor elements, initial pH of the medium, some organic solvents as precursor solvents, and precursors on their growth and digitoxin formation were investigated. Reduction of the basal CaCl₂ concentration by two-thirds promoted both digitoxin formation and shoot-differentiation while a threefold increase repressed these. Ca⁺⁺ proved to be responsible for this phenomenon. A threefold increase in the MgSO₄ concentration improved digitoxin formation, Mg⁺⁺ being responsible for this improvement. The optimal concentration of FeSO₄ coupled with Na₂-EDTA was similar to that in the MS basal medium. A threefold increase in the concentration of either KI, Na₂MoO₄ or CuSO₄ improved digitoxin formation, the optimal pH of the medium before autoclaving being 6.Among methanol, ethanol, propylene glycol, dimethyl sulfoxide and acetone as the precursor solvents, methanol and acetone were relatively the least inhibitive at 2%. Progesterone at 0.1mg/100ml medium improved the digitoxin content per flask to 180% of that of the control cultures, whereas no promotive effect from cholesterol was observed.

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Four isozymes of glucoamylase were isolated from a commercial preparation of the enzyme from Rhizopus niveus by ion-exchange chromatography on CM-Sephadex C-50 and/or the preparative isoelectric focusing method on Sephadex IEF.
The kinetic parameters, Michaelis constant Km and molecular activity k₀, for the hydrolysis of maltooligosaccharides (degree of polymerization DP from 2 to 7) were determined on the major component enzyme at pH 4.5 and 25°C. It was found that Km decreases and k₀ increases with increasing DP. From the DP-dependences of Km and k₀, the subsite affinities of the seven subsites of the enzyme active site were evaluated on the basis of subsite theory. These values are very similar to those obtained for an unfractionated preparation of Rhizopus delemar glucoamylase [Hiromi et al, B. B. A., 302, 362 (1973)].

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(±)-γ-Irones (1/9 mixture of cis-and trans-isomers and pure trans-γ-irono) were synthesized via the intramolecular Diels-Alder reaction starting from 2, 4-hexadienyl aniline derivative and β, β-dimethylacryloyl chloride.

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The cell wall of Conidiobolus lamprauges is composed of D-glucosamine, D-glucose and D-mannose in the ratio of 1:1:1. The first sugar originated from chitin and a part of the second sugar came from β-glucan.
Cell wall-associated, chitin-binding hemagglutinin (endo-CLA) was liberated from the cell wall with β-glucanase but not with chitinase, cellulase, protease, sodium chloride, 2-mercaptoethanol, hydrochloric acid, N-acetyl-D-glucosamine or chitooligosaccharides.
Endo-CLA and extracellular CLA (exo-CLA) showed almost the same sugar specificity and immunological properties in the hemagglutination inhibition test and the double immunodifFusion test, respectively.
Endo-CLA was detected in the cell wall before production of β-glucanase by this organism, whereas exo-CLA was produced in the culture filtrate simultaneously with the enzyme.
These results suggest that CLA was liberated from the β-glucan matrix in the cell wall by the action of β-glucanase.

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Many N-phenylsulfonylbenzamides were synthesized, and their biological activities were tested. Some of these compounds showed a high phytotoxic activity against barnyardgrass with no significant effect on rice plants at their germination stage. In particular, both N-allyl-2-chloro-N-phenylsulfonylbenzamide and N-allyl-2, 4-dichloro-N-phenylsulfonylbenzamide were the most active and a herbicidal test of these two compounds was done in paddy field conditions.

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Both the natural eldanolide [(3S, 4R)-(+)-3, 7-dimethyl-6-octen-4-olide] and its antipode, the pheromone produced by the male Eldana saccharina (Wlk.), were synthesized from optically pure methyl (R)-(+)-citronellate.

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Three multiple forms of lipoxygenase were isolated from rice embryos by DEAE-Sephacel chromatography. The pH-activity curves for the isolated enzymes differed from each other. They showed different products (positional) specificities for linoleic acid as substrate.

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Isoprothiolane sulfoxide, an oxidative metabolite of isoprothiolane, is easily reduced to isoprothiolane by the living rice plant. The reduction is completed within 2 days after addition to the culture solution. Reduction of the sulfoxide may be important in activating a possible propesticide, isoprothiolane sulfoxide, and in ensuring the residual effectiveness of the pesticide, isoprothiolane.

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Intraperitoneal injection of bestatin, a microbial aminopeptidase inhibitor, resulted in accumulation of acid soluble peptide-form amino acids in liver of fasting rats. This aminopeptidase inhibitor caused a slight increase in the rate of urinary excretion of acid soluble peptide-form amino acids. However, the excretion rates of only two amino acids of the acid soluble peptide-form were statistically higher in the bestatin injected rats than in the control rats.
Leupeptin, another microbial protease inhibitor, also resulted in peptide accumulation in liver of fasting rats. However, it was less effective than bestatin. Leupeptin did not show any effect on the rate of urinary excretion of acid soluble peptides in rats under the present experimental conditions.
These results suggest that bestatin sensitive protease (s) may participate in the process of protein degradation in the liver (and presumably in other tissues also), especially in the steps of the degradation of oligopeptides into amino acids.

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