Studies on the charactors of cocci especially on the orange colored strains isolated from rumen contents of fattening young British Friesian ox and Dexter cows were carried out. The organisms are round or oval, having 0.7 to 1.2 microns in diameter, appearing singly in pairs or in chains. Gram-positive, catalase negative, grow well on rumen liquid (RL)- glucose medium under strict anaerobic conditions. Optimum temperature lies between 35 and 40°C. Acid from glucose, mannose, maltose, sucrose, lactose, cellobiose, raffinose and starch. Lesser from arabinose and salicin. No acid from xylose, xylan and cellulose. The acid is almost exclusively L (+)- lactic acid. The most striking feature is their strong fermentability of raw starch, expressing their function in rumen. Including white strains isolated together, these organisms are considered very similar to Streptococcus bovis Orla-Jensen, however, as the lactic acid recovery from consumed glucose does not exceed from 40_??_50% in RL-glucose medium and 70_??_80% in polypeptone-yeast extract (PY)- glucose medium, the organisms can not yet be determined as true Streptococcus.
Orange color of the colored strains develops relating to the nitrogenous composition of culture media, such like orange in RL but no coloration in PY.

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Reversible transconformation of casein induced by heating at temperature from 15°C to 70°C and cooling to 15°C was studied using ultraviolet difference absorption spectra and pH measurement.
(1) Difference spectra of whole casein solution showed hyperchromisity (240_??_260 mμ and 283_??_310 mμ) and hypochromisity (260_??_283 mμ) resulting from red shift of absorption spectra which altered by the change of temperature. The difference spectra had 2 characteristic peaks at 287_??_288 mμ, due to tyrosine side chain, and 293_??_294 mμ, due to tryptophan side chain, which grew up with elevated temperature up to 70°C and disappeared after cooling.
(2) When κ-casein was heated at 60°C, light dispersion was caused by aggregation. After cooling casein solution became clear again.
(3) pH value of casein solution was changed by heating from pH 6.75 at 10°C to pH 6.54 at 50°C and pH 6.57 at 60°C, and by cooling from pH 6.59 at 60°C to pH 6.85 at 25°C and pH 6.76 at 10°C. These results suggest reversible transconformation of casein at elevated temperature.

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The initial action patterns of saccharifying α- and liquefying α-amylase of Bacillus subtilis on various β-limit dextrins were investigated and the following results were obtained:
1) The initial attack of saccharifying α-amylase occurred much more readily at the third or more inner bond toward the reducing end from the branching point.
2) In the case of liquefying α-amylase, the amylase seemed to hydrolyze the glucosidic linkage at the fifth or more inner bond from the branching point.
These observations were quite comparable with the results obtained by using linear maltooligosaccharides in the earlier report^*, where the glucosidic linkages to be split by these amylases are determined by the certain distance from the non-reducing end of the saccharides.

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The microbial transformation of DL-5-(β-methylmercaptoethyl)-hydantoin (DL-Hyd) to L-methionine was studied. The bacteria which can convert DL-Hyd to methionine and carbamylmethionine was isolated from soil and identified as Bacillus coagulans. Among eleven strains of B. coagulans and other bacteria from IAM culture collection, none of the bacteria was capable of forming L-methionine from DL-Hyd. Methionine and carbamyl-methionine formed from DL-Hyd were identified as L- and D-form, respectively.
The optimal conditions for producing L-methionine were studied, and the yield of Lmethionine and carbamyl-D-methionine attained to 56.7 and 17.6 molar percent respectively, when the bacteria was incubated aerobically at 30°C for 26 hr using the following medium: 1% of starch, 0.25% of urea, 0.8% of corn steep liquor, 0.5% of KH₂PO₄, 0.01% of MgSO₄ and 0.001% of each Fe and Mn sulfates.
The mechanism of transformation of DL-Hyd to L-methionine and carbamyl-D-methio-nine was discussed.

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When a crude enzyme produced by Aspergillus sojae No. 48 in wheat bran culture was tested for breakdown of various kinds of parenchymatous tissues, such as potato, carrot, radish, spinach, cabbage, onion, garlic, rice, soybean, and some fruits, it was recognized that the enzyme degraded cell wall cementing material in middle lamella and caused maceration of these tissues. Cell wall of separated cells was further decomposed by the enzyme.
Also in this paper, some properties of cellulolytic enzymes contained in the crude enzyme preparation were investigated and discussed. At least, three components; cellulase C₁ (filter paper disintergrating activity), cellulase C_x (CM-cellulase) and β-glucosidase activities were involved and isolated by gel filtration on Sephadex G-200. pH optimums of cellulase C₁ and cellulase C_x were both pH 5.0, on the other hand, β-glucosidase was pH 4. 5. Cellulase C₁ and C_x were found to be stable at the range of pH 3_??_8, while β-glucosidase was quite labile below pH 3. These cellulases were completely inactivated by heat treatment at 70°C for 10min.

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In the previous paper, it was reported that the enzyme produced by Aspergillus sojae No. 48 could degrade various kinds of parenchymatous plant tissues, and at least three components of cellulase were contained in it. The present paper concerns with further studies of the other enzyme activities in a crude enzyme preparation produced by this mold. When macerating activity was measured by the weight loss of potato tuber, optimum pH of this enzyme was found to be 5. 5. Concerning the pectolytic enzymes, there were pectic acid saccharifying, liquefying and pectin liquefying activities, but no pectin esterase activity. Except these hydrolytic pectinases, polyglacturonide lyase which eliminated pectin, but not pectic acid was also found in this enzyme. By the results obtained in the effect of pH on enzyme activities, it was assumed that the enzyme which reduces the viscosity of pectin but not of pectic acid was the enzyme causing maceration. Hemicellulases which hydrolyzed xylan and araban, β-glucanases which hydrolyzed laminarin, yeast glucan, pachyman and lutean, chitinase and proteases activities were also found in the enzyme preparation.

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Cultural conditions for the polysaccharide production by Bacillus polymyxa No. 271 were studied. The strain was capable of producing an acidic polysaccharide from xylose, fructose, glucose, mannose, galactose, lactose, maltose, glycerol and mannitol. The constituent sugars and some physical properties of the polysaccharides obtained from the above sugars were essentially identical.
The most suitable medium contained 4% glucose, 0. 5% peptone (pancreatic digest of casein), 0.5% calcium carbonate and other inorganic salts. Monobasic potassium phosphate, magnesium sulfate and manganese sulfate were required. Both Mg²⁺ and Mn²⁺ were essential for the formation of polymer. In order to maintain the pH value of 5.5 or above during the fermentation, the addition of calcium carbonate was effective. The optimum initial pH value and temperature were 6.5 and 28°C, respectively. In these conditions, the polysaccharide produced from glucose was obtained in a yield of 16.5 gram per liter corresponding to 45%, based on the consumed sugar.

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During the germination of barley activities of protein SS reductase and proteolytic enzymes were determined in relation to the activation of zymogen β-amylase. Protein SS reductase was assayed amperometrically titrating SH groups increased by the enzymatic decomposition of barley protein at pH 7.0 and 30°C for 130min. The activity was hardly detected in ungerminated barley but was found to increase rapidly in the early stage of germination. The proteolytic activity was determined using gelatin, haemoglobin and casein as the substrates. The casein hydrolyzing activity during the germination of barley increased rather slowly than the protein SS reductase activity. In contrast to the increase in these enzymatic activities zymogen β-amylase decreased with the advance of germination days. It was also found that salt-soluble zymogen β-amylase obtained from ungerminated barley was activated in vitro by the protein SS reductase from malt in the presence of coenzymes.
From these results above and the formerly reported results on activation of zymogen β-amylase with thiols and proteolytic enzymes, it was postulated that the activation of zymogen β-amylase during the germination of barley would be concerned with the activi-ties of the protein SS reductase and the malt proteinase.

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A β-mannosidase was extracted from the Koji culture of Rhizopus niveus and purified by DEAF-and SE-Sephadex column chromatographies and Sephadex G-100 gel filtration.
The purified enzyme was free from mannanase. The pH optimum of the enzyme was 5.0 and the temperature optimum was 40
C. The initial rate of hydrolysis of five mannooligosaccharides by the enzyme was in the following order,
mannotetraose (M₄)=mannotriose (M₃)>mannobiose (M₂)>mannopentaose (M₅)>mannotexaose (M₆).
When M₂, M₃ and M₄ were used, in the concentration of 10^-2M, as a substrate, M₃, M₄ and M₅ were detected respectively by paperchromatographic separation of the reaction products.

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The amount of sorption water in various starch granules which equilibrate with different water vapour at 25°C, and 40°C were measured. Isosteric sorption heat was calculated by Clausius Clapeyron equation.
1) Hysteresis loop area in sorption and desorption of potato starch was the largest and diminished in order of tapioca, waxy rice, sweet potato and rice at 25°C. These values at 25°C were higher than at 40°C.
2) Unilayer sorption water in various starch granules was calculated by B. E. T. equation were approximately 7.7% (dry basis, 25°C).
3) Isosteric sorption heat was almost constant at more than about 20% sorption water.

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Three Penicillium strains having the excellent ability to produce indole alkaloids in surface and submerged cultures were isolated from the air of the circumferance of laboratory. These strains were all identified as Penicillium concavo-rugulosum ABE.
By selection of the strains and modifying the medium, the total alkaloid yield in surface cultures could be increased up to ca. 1000mg alkaloids per 10 cultures (culture filtrate 1.35 liter, and the mycelium 15g, based on dry weight).
From these cultures, a hitherto known ergot alkaloid chanoclavine-(I) and, further, two new interconvertible alkaloids, rugulovasine A and B, have been isolated in good yields.
Rugulovasine A crystallized from benzene as colorless prisms; C₁₆H₁₆O₂N₂, mp 138°C (decomp.), [α]²²_D 0°, [α]²²₅₄₆ 0° (c=0.5, pyridine). Rugulovasine B separated from benzene as colorless resinous oil; C₁₆H₁₆O₂N₂, [α]²²_D 0°, [α]²²₅₄₆ 0° (c=0.5, pyridine). It yielded crystalline salts: the hydrochloride, C₁₆H₁₆O₂N₂•HCl•H₂O, white prisms from water, mp 187°C (decomp.), [α]D 0° (c=0.5, pyridine); the oxalate, C₁₆H₁₆O₂N₂•1/2 C₂H₂O₄, colorless prisms from water, mp 217°C (decomp.), [α]₂₂ 0° (c=0.5, pyridine).
Both rugulovasine A and B gave reddish purple color, turning into bluish purple color with Allport and Cocking's reagent. They were reversibly converted into each other in various organic solvents, especially in alcohols. They were considered to be the chanocla-vine type of new ergot alkaloid isomers, containing an opened D ring and an unsaturated lactone ring in their molecules.
The rugulovasines have been found with chanoclavine-(I) also in the cultures of various molds, such as Penicillium rugulosum, Corticium caeruleum, Pellicularia filamen-tosa, etc., besides Penicillium concavo-rugulosum.

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Microbiological conversion of Borneol was studied by newly isolated pseudomonado (strain H) as a link of the series of microbiological conversion of terpenes. Borneol fermentation was divided into three stages by time course change. Neutral main metabolite of each stage was identified as camphor, campholytolactone, 2, 6-diketocamphane. So the metabolic pathway of camphor by strain H seems to take different pattern as reported previously.

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This report deals with a gaschromatographic method for the identification of benzofuran derivatives. Shimadzu Model GC-1 C with hydrogen flame ionization detector was used in this study. The useful information can be obtained from suitable selection among various column packings and operating conditions tested.

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A method for identification of benzofuran derivatives was developed employing ascending thin-layer chromatography over silica gel and silica gel-cellulose with various solvents as a developer. The results obtained are listed in Table I and II.

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