A Jar fermentor technique used for microbes is applied to a large-scale culture of tobacco cell suspension in both batch and semi-continuous culture systems.
A 30 liter fermentor contained 20 liters culture which, after 5 days incubation at 28°C, yielded a suspension of 20g/liter as dry weight and 650ml/liter as packed cell volume in a batch system. Growth curves of tobacco cell suspensions showed an optimal mean doubling time of 1 day during an exponential growth phase of 1_??_4 days. In the semi-continuous cultures, the yields of dry weight were 12_??_13g/liter day.

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Succinylchymotrypsin was prepared from succinylchymotrypsinogen with trypsin and purified by DEAE-Sephadex chromatography.
The total 15 lysyl and N-terminal amino residues of chymotrypsinogen were succinylated. About 23.5 out of 49 seryl and threonyl residues and none in the total 4 tyrosyl residues in succinylchymotrypsin were modified
The helix content of succinylchymotrypsin was about 8.4%.
The pK_app of N-terminal amino group in succinylchymotrypsin did not change and that of tyrosyl residues shifted to alkali side, in comparison with those of unmodified α-chymotrypsin.
The k_cat of succinylchymotrypsin increased and its Km did not change within experimental error, in comparison with those of α-chymotrypsin.

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A new method is described for the synthesis of pyrimidine nucleosides via fully acylated sugars in the presence of Lewis acids.

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Separation and determination of the optical isomers of chrysanthemic acid has been carried out by gas chromatography. The diastereoisomeric esters of chrysanthemic acid with l-menthol were separated on a column of 2% QF-1 coated on Chromosorb W. d-trans, l-trans and dl-cis Chrysanthemic acids were resolved but d-cis and l-cis acids were not separatable from one another on any column tested. These isomers of chrysanthemic acid were not isomerized during esterification and gas chromatographic operation, and their ratios were determined from their peak area ratios. The l-bornyl esters of the isomers of chrysanthemic acid were not so easily separatable as the l-menthyl esters.

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Kluyvera citrophila KY 3641 cultivated aerobically more than 48 hr produced peicillin acylase. Sucrose, monosodium glutamate and aspartic acid stimulated the growth of the cells, whereas glucose, fructose, maltose and lactose inhibited the growth and acylase production. Enzymic reaction was carried out with whole broth, too, instead of using separated intact cells. The cells maintained its acylase activity more than one month and could be used repeatedly. Acetone-dried or freeze-dried cell was also available for enzymic reaction. Identification of 6-APA was also described.

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The persistence of thiophanate-methyl, dimethyl 4, 4'-o-phenylenebis (3-thioallophanate), and its metabolites on plant leaves and glass plates was examined using a radiolabeled compound. Apple and grape leaves and glass plates treated with thiophanate-methyl-phenyl-¹⁴C were kept outdoors, except on rainy days and nights when they were kept in green house.
Half lives of thiophanate-methyl on apple and grape leaves were around 15 and 12 days, respectively. On glass plates half life was around 3 days. The per cent of abundance of the parent compound and a major degraded product, methyl 2-benzimidazolecarbamate, versus applied thiophanate-methyl at 14 days after treatment was as follows; 52.6 and 10.1% on apple leaves, 49.5 and 8.9% on grape leaves, and 5.5 and 24.1% on glass plates, respectively.

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Amino acid composition of the CGMMV^* coat protein was determined to be as follows: Asp₂₀, Thr₁₀, Ser₂₄, Glu₁₀, Pro₆, Gly₉, Ala₂₁, Val₇, Ile₇, Leu₁₈, Tyr₄, Phe₉, Lys₄, His₁, Arg₈, Trp₂. No terminal α-amino group was detected by dinitrophenylation method. The carboxyl-terminus was found to be serine by hydrazinolysis of the protein and digestion with carboaypeptidase A.
For sequence analysis of the coat protein, tryptic digestion was accomplished at pH 8.0 resulting in ten soluble and several insoluble peptides at pH 4.5. The amino acids contained in soluble peptides accounted for 91 out of 160 residues in the whole protein. The amino acid sequences of ten soluble peptides were determined.
From the similarities of amino acid sequence of the peptides to those of TMV^* protein, CGMMV was assumed to be a strain of TMV group.

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The yolk index of newly laid egg was 0.50, while that of the egg stored at 30°C for 15 days in an atmosphere of carbon dioxide at a pressure of 2kg/cm₂ and in air was 0.43 and 0.25, respectively.
The carbohydrate content of ovomucin gel (B) obtained from the eggs stored in an atmosphere of carbon dioxide was higher than that of ovomucin gel (B) obtained from the eggs stored in air.
The free boundary electrophoretic pattern of ovomucin gel (B) obtained from the eggs stored in an atmosphere of carbon dioxide consisted of two peaks, and the relative mobility of each peak showed the same value as that of the corresponding each peak of ovomucin gel (B) obtained from newly laid egg white.
The fractionation pattern obtained by density gradient column electrophoresis of ovomucin gel (B) obtained from the eggs stored in an atmosphere of carbon dioxide showed two peaks, peak-F and peak-S, while that of the eggs stored in air showed a considerable diminution in peak-F.
From these results, discussion was made about the occurrence and mechanism of egg white thinning when eggs were stored in an atmosphere of carbon dioxide.

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The volatiles, produced from casein during roasting at 250°C for 1 hr, were fractionated and analyzed by various methods including the combination of gas chromatography and mass spectrometry. The following kinds of compounds were tentatively or conclusively identified: six aliphatic aldehydes, benzaldehyde, acetophenone, hydrogen sulfide, methanethiol, dimethyl disulfide, five primary alkylamines, piperidine, N-ethylcyclohexylamine, two alkylpyrazines, ethylmethylpyridine, three aliphatic carboxylic acids, pyruvic acid, benzoic acid, six aliphatic alcohols, and three aromatic hydrocarbons. α-Dicarbonyls were not formed in any detectable amount.
The results obtained above are discussed in comparison with those of pyrolysis of some kinds of α-amino acid and also with those of some roasted foods.

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A small jar fermentor was developed in order to investigate the effect of oxygen supply on hydrocarbon fermentation. Several indices to oxygen transfer were examined with this small jar fermentor. Conditions for suitable oxygen supply were examined in L-glutamic acid fermentation from hydrocarbon by use of shaking flasks and these small jar fermentors. The data indicated that the rate of oxygen transfer ought to be more than 14.3×10^-7mole/ ml•min in order to obtain satisfactory results. The coefficient of oxygen transfer rate (K_La/H) decreased as the fermentation went on, so the supply of oxygen enriched gas mixture was effective to increase the production of L-glutamic acid.

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γ-L-Glutamyl-L-methioninesulfoxide was isolated from green gram seed. The peptide, isolated from plant material for the first time, was present in large amounts in seeds, but was not detected in commercially obtained etiolated seedlings as was the case with other γ-glutamyl peptides.

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Some conditions of autolysis in cultured tobacco cells were examined for temperature, cell culture age and aeration. Cells autolyzed readily at 45°C. Seventy percent of the dry matter, almost 100% of the soluble sugar, 40% of the insoluble sugar and 60% of the total nitrogen in the initial cells were excreted within 5 hr of incubation in water. At lower physiological temperatures, excreted substances were reabsorbed into cells during the early period of incubation under aerobic conditions.
Rapidly growing cells excreted larger amounts of sugar, nitrogen and solid matter than did non-growing cells during autolysis at 30°C.
Plasmolysis was observed in autolyzed cells.
Autolysis was makedly stimulated by anaerobic conditions.

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Two new juvenile hormone analogues with _??_ cvclopentane (IVa) or cyclohexane (IVb) rings at the terminal position were prepared and were shown to possess high juvenile hormone activity.

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Two roosters of Singles Comb White Leghorn breed were fed on a formula feed containing 2-¹⁴C glycine for 16_??_17 days. The animals were killed and the specific activities of glycine in the tissue proteins and the purine ring of the excreted uric acid were measured. The amount of synthesized glycine in the rooster was calculated by the dilution method based on the specific activities of glycine in the liver protein and the absorbed one. The rooster absorbed about 1g of glycine and synthesized about 10g of glycine per day. Quantitative aspect of glycine metabolism in the rooster was discussed.

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A mannan-hydrolyzing enzyme produced by _??_ certain strain of Bacillus subtilis was purified from the culture broth and isolated in a crystalline state by being treated with several ion-exchangers. The optimal pH of the enzyme was 6.0. The enzyme was stable in a pH region of 5.0 to 9.5 and at temperatures less than 55°C. The enzyme attacked only β-1, 4-mannosidic linkages in the main chain of galactomannan of soybean seed coat, guar, gum and coffee bean, and of glucomannan of konjak (Amorphophalus konjac). Investigation of the hydrolysis mode revealed that the enzyme attacked coffee bean galactomannan endowise to form mannobiose, mannotriose and mannotetraose. The action patterns on several mannohomooligomers prepared from a partial hydrolysate of coffee bean galactomannan were also investigated, indicating that the enzyme preferentially attacked the β-1, 4-mannosidic linkages that were present apart three to four mannose residues from the non-reducing end of the mannose chain.

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Two aldehydic C₂₀-gibberellins, L-2 and L-4, were isolated from the immature fruits of yellow lupine (Lupinus luteus L.). L-2 was shown to have the structure II and named gibberellin A₂₃. L-4 was identified as gibberellin A₁₉ (VI). Two new C₂₀-gibberellins, tentatively called 3, 13-dihydroxy GA₁₅ (IV) and 13-hydroxy GA₁₅ (VIII), were derived from gibberellins, A₂₃ and A₁₉, respectively. The biological activities of four 3, 13-dihydroxy C₂₀-gibberellins-GA₁₈ (I), GA₂₃ (II), GA₂₈ (III) and 3, 13-dihydroxy GA₁₅(IV), which were isolated from the fruits except for 3, 13-dihydroxy GA₁₅-were compared in six gibberellin bioassays.

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A novel lignan, named phrymarolin-I, has been isolated from Phryma leptostachya L. The structure (I) for phrymarolin-I represents the first example of a lignan characterized by a 1, 2-dioxygenated-3, 7-dioxabicyclo [3. 3. 0] octane ring. Its stereochemistry was determined from NMR spectra.

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β-Ionone (I) was oxidized to 2, 3-epoxy-β-ionone (II), which was converted to 2, 3-dihydroxy-β-ionone (III) by acid treatment. III was reduced to 4-(1, 2-dihydroxy-2, 6, 6-trimethylcyclohexan-1-yl)-2-butanol (V), which was converted, by oxidation, to cis- and trans-theaspirone (1-oxa-8-oxo-2, 6, 10, 10-tetramethyl spiro-(4, 5)-6-decene) (VII-A), (VII-B) and dihydroactinidiolide (2-hydroxy-2, 6, 6-trimethylcyclohexyliden-1-acetic acid lactone) (IX).

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Protoplasts were isolated from cultured tobacco cells by removing the cell wall enzymatically. We examined the time courses of treatment with some different concentrations of enzymes (cellulase and macerozyme) and with some different kinds and concentrations of sugars (sucrose, sorbitol and mannitol) which produce good conditions for protoplast isolation. The best conditions for protoplast preparation from tobacco cells cultured in vitro were: use of actively growing cells and isolation of protoplasts with 5% cellulase and 0.1% macerozyme in 0.5% sorbitol.

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