Among 843 strains of 56 microbial genera tested, those producing considerably high elastolytic activities in the culture broths belonged to only 5 genera, Bacillus, Brevibacterium, Flavobacterium, Pseudomonas and Streptomyces. When Flavobacterium sp. No. 9-35 which was isolated from soil, was cultured in a commercially applicable medium, 155 units per ml of elastolytic enzyme was produced which was about 4 fold higher than that produced with a complex medium by “the best” strain of those reported.
Twenty six-fold purified enzyme preparation from Flavobacterium sp. No. 9-35 showed the optimum temperature of 40°C and the optimum pH of 7.5 with phosphate buffer for the activity. The enzyme was stable at neutral pH and at less than 40°C, while the complete inactivation was observed at 60°C for 20min. CuSO₄, ZnSO₄ and HgCl₂ strongly inhibited the activity, wheras p-hydroxymercuribenzoate and EDTA did not. 0.1M NaCl inhibited the activity 45% at pH 8.5, while activated it 20% at pH 7.2. The enzyme was strongly adsorbed on elastin at pH 8.5.

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A Chlorella strain with high growth rates under heterotrophic growth condition was isolated from a pond and identified as Chlorella regularis. The optimal temperature for growth was 36°C and the optimal pH 6.0_??_7.0. Glucose, galactose, acetic acid, ethanol, acetaldehyde and pyruvic acid may serve as the carbon source supporting the growth under both dark and light conditions.
The specific growth rate μmax was; 0.3 hr^-1 under autotrophic condition, 0.28 hr^-1 under heterotrophic condition, and 0.45 hr^-1 under mixotrophic condition. In the dark heterotrophic culture, the yield of the cells to glucose was 0.5_??_0.6, that to acetic acid 0.48, and that to ethanol 0.66. When dark heterotrophic cells were cultured under mixotrophic condition, the yield to glucose increased to 0.82, that to acetic acid to 0.85 and to ethanol to 0.91.
The protein and chlorophyll contents of the cells in autotrophic culture were approximately, 60% and 4Y., respectively. These contents of the cells in dark heterotrophic culture changed in accordance with the growth phase. The protein of the cells reached to, approximately, 60% and the chlorophyll to 2% in stationary phase of growth, although the cells were maintained even in the dark.

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The formation of Suzukacillin A (SK-A) a peptide antibiotic containing α-amino isobutyric acid (α-AiB) as an unnatural amino acid component, was promoted with increased supply of α-AiB while alternatively affected by the supplied amounts of L-valine; promotively with its adequate amounts (0.05_??_0.2%) but inhibitorily with its excess amounts (0.5_??_1.0%). At the incorporation tests with ¹⁴C-labeled pyruvate or L-valine, the radioactivity was incorporated into SK-A and mainly into α-AiB of SK-A formed, where a quantitative dilution occurred in the coexistence of their cold preparations. This suggests that L-valine or its related metabolite is a precursor of α-AiB in the biosynthetic process. Also the same was the case in the incorporation test with ¹⁴C-labeled α-keto isovalerate which seemed likely to be less effective precursor than L-valine on the basis of the radioactivity incorporated into α-AiB of SK-A peptide. When L-valine-1-¹⁴C or pyruvate-1-¹⁴C was used as incorporation substrate, no radioactivity in α-AiB of SK-A formed was found, and thus in the biosynthetic process of α-AiB from L-valine, 1-C of L-valine molecule might be decarboxylated and ruled out of α-AiB molecule. From the results obtained at the tests with each of radioactive pyruvate, L-valine and α-AiB, concomitantly supplied with various amounts of cold α-AiB or L-valine, it was suggested that there may occur a certain competitive correlationship between α-AiB and L-valine in the mobilizing process to SK-A peptide synthesis. The role of L-valine played in the formation mechanism of SK-A peptide was signified and discussed, where a presumptive scheme for the formation mechanism of SK-A peptide was tentatively presented.

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Rhodotorula glutinis K-24 produces a highly viscous polysaccharide in an acidic medium. The polysaccharide was separated from the supernatant of the culture broth by addition of acetone, and partially purified by ethanol precipitation. The partially purified polysaccharide, of which the hydrolysate gave three spots corresponding to fucose, galactose, and mannose on a thin-layer chromatoplate, seemed to be homogeneous in free boundary electrophoresis, but in ultracentrifugation, it gave two peaks, major and minor, on the sedimentation pattern. The minor component containing mannose was removed by the fractional precipitation with ethanol. The purified polysaccharide thus obtained was found to be a fucogalactan ([α]²⁵_D-38°) composed of L-fucose and D-galactose in a molar ratio of about 1:1. Partial hydrolysis revealed the presence of core galactan in it. The sedimentation velocity of the purified polysaccharide decreased rapidly with increasing concentration of polysaccharide above 0.5%. The sedimentation coefficient at zero concentration was 10.7×10^-13 at 30°C in 1% NaCl aq.

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An alkalophilic proteinase was isolated in a crystalline state from the culture filtrate of a Streptomyces sp. which was screened as an organism to grow on the media of pH values above 9.5. The proteinase was inactivated by DFP. It was also inactivated by EDTA at temperatures above 25°C but, was not inactivated by the reagent at temperatures lower than 20°C. The proteinase showed its optimal pH at 12 or higher than 12 and the proteolytic action occurred even in solutions containing 0.2M sodium hydroxide. The enzyme exhibited its maximum activity at 60°C.
The present paper employs a new definition for a unit of the so called proteinase activity.

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The nature of the commercial smoke condensates (wood vinegar liquors) obtained from six kinds of wood was studied in relation to their flavors.
Undistilled smoke condensates had an unpleasant resin-like odor, while distilled ones had a strong pungent smoky flavor mixed with cresolic and furfural-like doors. These differed slightly with various kinds of wood materials. Among the six kinds of wood materials, oak B^* and bamboo gave organoleptically the most acceptable smoke flavors.
The amounts and compositions of flavor components in the carbonyl, non-carbonyl neutral, basic, acidic and phenolic fractions contributed to the differences in aroma. In particular, the composition of the phenolic fraction was thought to be important.

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Carbonyl, non-carbonyl neutral and basic fractions obtained from commercial aqeuous smoke condensates were separated by GC and/or silica gel column chromatography.
Ninety-eight constituents were identified (thirty-one compounds were not previously reported in smoke condensate) and twenty-two were tentatively identified on the basis of tR of GC, IR, MS (GC-MS) and NMR data.
The yields of all the smoky aroma constituents (four aliphatic alcohols, five aliphatic ketoalcohols, six aliphatic ketones, twenty cyclic monoketones, three cyclic diketones, ten acids, two esters, twelve lactones, nine furan derivatives, six N-compounds, four aromatic hydrocarbons, two aromatic alcohols, nine aromatic carbonyls, twenty-four phenols and phenol ethers and five pyrocatechols) detected in commercial wood vinegar were estimated.

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An improved method of the purification of ricin D was investigated. Ricin was purified by gel-filtration through Sephadex G-75 at pH 8.0, followed by either CM-cellulose column chromatography at pH 6.5 or DEAE-cellulose column chromatography at pH 8.5. The homogeneity of the purified ricin was criticized by polyacrylamide gel disc electrophoresis.The purified ricin behaved homogeneous also in ampholine electrophoresis, indicating the isoelectric point of 7.34. Ricin thus purified was identical with ricin D in electrophoretical migration and toxicity. By the measurement of optical rotatory dispersion of the purified ricin, ORD constant, λc, Moffitt-Yang parameters, a0 and b0, were evaluated to be 235nm, -138 and -66, respectively.

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During the growth of hydrocarbon-utilizing bacteria, Arthrobacter roseoparaffineus KY 4301 and Microcoecus paraffinolyticus KY 4306 on n-alkane as the sole source of carbon, it was found that a considerable amount of an adenine ribonucleotide accumulated in the medium. The nucleotide was isolated in crystalline form from the broth culture through the chromatography on ion-exchange resins and Sephadex G-10 columns. It was identified as cyclic adenosine 3', 5'-monophosphate by paper chromatography, ultraviolet-absorption spectra, infraredabsorption spectra and other chemical analyses.
For the production of cyclic adenosine 3', 5'-monophosphate, n-alkane, particularly n-dodecane or n-tetradecane, was the most preferable carbon source under the culture condition in this work. With n-tetradecane, approximately 1.4g per liter and 0.5g per liter were produced respectively by Arthrobacter roseoparaffineus KY 4301 and Micrococcus paraffinolyticus KY 4306 under the same condition.

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The specific activities of aspartokinase, threonine deaminase, ä-acetolactate (AL-) synthetase and transaminase B were measured for the cell-free extracts of E. coli auxotrophs and E. coli C-6 (wild strain) grown on fermentation medium containing 5% glucose as the carbon source. The properties of each enzymes in crude extracts were also investigated. In a threonine producer, strain No. 15 (Met-), the lysine- or methionine-sensitive aspartokinase which is insensitive to feedback inhibition of L-threonine was derepressed about five folds when the auxotroph was cultured in the presence of a limited amount of methionine.
The formation of aspartokinase, threonine deaminase and transaminase B were increased in the presence of L-valine, while the presence of L-isoleucine appreciably repressed aspartokinase, threonine deaminase and transaminase B in the strain No. 234 (Met-, Val-) and strain T-3 (Met-, Val-, Ileu-).
From the above results, the mechanisms of L-threonine production in E. coli auxotrophs were discussed. The mechanisms of temperature-shift effect in E. coli No. 234 were also proposed, based on the results of the changes of enzyme activities during the fermentation.

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The effect of an antibiotic, borrelidin, on L-threonine fermentation by E. coli auxotrophs were studied and it was found that the production of L-threonine was greatly increased by the presence of borrelidin. The production of L-threonine was considerably increased by the addition of L-aspartic acid together with borrelidin. The maximum amount of L-threonine accumulated by E. coli No. 234 was 15.0mg/ml in the medium containing 50mg/ml of glucose and 5mg/ml of sodium-aspartate.

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The effect of organic acids on the production of macrolide antibiotics by a strain of S. venezuelae MCRL-0376 was studied. Organic acids such as, citric, succinic, acetic, malic, malonic and fumaric acid etc. stimulated antibiotic production, while propionic acid which is reported to be incorporated into an aglycone of macrolide antibiotics showed inhibitory effects on the growth and antibiotic production. However, in the presence of lipoic acid, propionic acid acted as other organic acids.
Addition of organic acids in the medium caused the accumulation of narbonolide, an aglycone of narbomycin, by S. venezuelae MCRL-0376, though this compound was not produced in the absence of organic acids.
Narbonolide did not inhibit the growth of Bacillus subtilis, but by incubation with washed cell of S. venezuelae MCRL-0376, yielded an antibiotic substance (mainly picromycin) active against this organism. Further, addition of narbonolide in the culture of strain MCRL-0376 enhanced the antibiotic production.

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A crude inhibitor for pancreatic lipase was extracted from soybean seeds. The lipase activity decreased curvilinearly with an increase in inhibitor concentration. At a low inhibitor concentration, enhanced inhibition was observed by the co-existence of protein such as bovine serum albumin in the reaction mixture. The lipase activity was inhibited immediately after the addition of inhibitor which did not cause the significant destraction of substrate emulsion. The lipase activities of Aspergillus niger, Rhizopus delemar and castor bean seeds were also inhibited. The inhibition was observed when various oil substrates such as soybean oil, linseed oil, olive oil emulsions and Ediol were used, and the extent of inhibition varied among them. Column chromatography of inhibitor on Sephadex G-100 showed that the molecular weight of a main peak of inhibitor was estimated as about 80, 000.

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The specificities of the alkaline and neutral proteases from the marine-psychrophilic Pseudomonas sp. No. 548 were investigated using oxidized insulin B-chain and various synthetic peptides as substrates. The alkaline protease was able to cleave the peptide or amide bonds containing a carboxyl group of amino acid residues, such as L-alanine, L-glutamic acid, L-arginine, L-leucine, L-phenylalanine, L-tryptophan, L-tyrosine, and L-valine. On the basis of the specificities, the enzyme might be classified as a serine protease, comparing the specificities of the proteases so far reported, although the enzyme is inactivated with ethylenediamine tetraacetic acid as well as diisopropylfluorophosphate.
The neutral protease catalized the hydrolysis of the peptide bonds containing an amino group of hydrophobic amino acid residues, such as L-isoleucine, L-leucine, and L-phenylalanine. These specificities agree closely with those of the neutral proteases which have been reported.

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An NAD linked formate dehydrogenating enzyme which catalyzed the last step of methanol oxidation system was extracted from the methanol-grown Kloeckera sp. No. 2201. The specific activity of the enzyme in the extract of methanol-grown cells was found to be considerably higher than that of the glucose-grown cells. The enzyme was purified about 35-fold from the extract of methanol-grown cells by heat treatment, column chromatographies on DEAE-cellulose and on hydroxylapatite, and Sephadex G-200 gel filtration. The purified enzyme was shown to be homogeneous by analyses with electrophoresis and ultracentrifugation. The purified enzyme was a kind of NAD: formate oxidoreductase (EC, 1. 2. 1. 2) which catalyzed specifically the oxidation of formate to carbon dioxide. The Km values were 22mM for formate and 0.1mM for NAD. The enzyme was inactivated by potassium cyanide, sodium azide, and p-chloromercuribenzoate but not by any metal-chelating reagents tested. Other general properties of the enzyme were also investigated.

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Human casein was separated by gel filtration on a column of Sephadex G-200 with 0.1M Tris buffer (pH 8.5) containing 1.0M NaCl. The effluent which increased in turbidity at 25°C was centrifuged at 25, 000×g for 30min and the precipitate was obtained as Fraction 6. After centrifugation, the effluent was separated into 5 elution fractions.
Disc gel electrophoretic patterns of each fraction showed occurrence of secondary bands other than major bands especially in Fractions 3, 4 and 5. The casein solutions unheated and heated at 100°C for 5 and 10min were kept at 5°C for 5 days. No marked changes of electrophoretic pattern were observed among these casein solutions. However, when a casein solution heated at 100°C for 5min was chromatographed under the same condition, secondary bands also appeared.

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The stereochemistry of fukiic acid (I) was elucidated. The relative configuration was determined to be erythro by IR spectrum and the evidence of acetonide formation. The absolute configuration of fukiic acid was assigned as S-configuration at C-2 and R-configuration at C-3, by Horeau's method and the identification of ozonolysis product to be hibiscus acid (10 a).

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To establish a novel process for the economical production of alkaline protease from n-paraffins by fungi, the variety of fungi were screened for the production of alkaline protease from n-paraffins and Fusarium sp. S-19-5 was selected as an excellent strain, and then the attempt was made to obtain some mutants capable of producing alkaline protease in high yield. A kabicidin resistant mutant No 5-128 B, was successfully obtained from Fusarium sp. S-19-5 by treatment with ultraviolet light. This mutant No 5-128 B showed high productivity of alkaline protease, about ten times that of the parent strain. The morphological characteristics and growth response to Kabicidin of the mutant No 5-128 B were compared with those of the parent stain.

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To obtain new plant growth retardants, about fifteen trimethylammonium compounds were prepared from terpenoid aldehydes or ketones and were bioassayed. N, N, N-Trimethyl-1-methyl-3-(2', 6', 6'-trimethyl-2'-cyclohexen-1'-yl)-2-propenylammonium iodide (35), hitherto unknown, was the most effective growth retardant, the activity being far superior to that of AMO-1618.

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Lipogenesis in the fatty liver of rat, which was induced by feeding an amino acid imbalanced diet containing 8% casein supplemented with 0.3% DL-methionine, has been studied by measuring the incorporation of glycerol-1-¹⁴C, palmitate-1-¹⁴C, citrate-1, 5-¹⁴C, pyruvatel-¹⁴C and pyruvate-2-¹⁴C into various lipid fractions and ¹⁴CO₂ during in vitro incubation of liver slices.
The total radioactivity of liver lipid per 100g of the body and the incorporation of each substrate into triglyceride in the lipid were significantly higher in the imbalance group than the control group. Conversion of each substrate to ¹⁴CO₂ was not imparied in the imbalance group.
It is evident from these results that the induction of this type of fatty liver is due mainly to the triglyceride synthesis by both the fatty acid synthesis and the transesterification of fatty acid.
These results are considered to support the previous assumption in which acetate-1-¹⁴C was used as a precursor.

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Mutants resistant to phenylalanine- or tyrosine-analogs were isolated from a wild type strain of Corynebacterium glutamicum ATCC 13032 and a tyrosine auxotroph TL-3 by mutagenic treatment with N-methyl-N'-nitro-N-nitroso guanidine (NTG) and screened for L-phenylalanine production. The growth of C. glutamicum ATCC 13032 was strongly inhibited by a phenylalanine analog, p-fluorophenylalanine (PFP), at a concentration of 1mg/ml. C. ghutamicum TL-3 was inhibited by PFP and another phenylalanine analog, β-2-thienylalanine (TA), at a concentration of 50μg/ml and 400μg/ml respectively. A prototrophic mutant resistant to PFP (4mg/ml) produced L-phenylalanine at a concentration of 5.5mg/ml and a trace amount of L-tyrosine in a cane molasses medium containing 10% of sugar calculated as glucose. A tyrosine auxotrophic mutant resistant to PFP (100μg/ml) and p-aminophenylalanine (PAP) (1mg/ml), 31-PAP-20-22, produced L-phenylalanine at a concentration of 9.5mg/ml in the molasses medium. L-Phenylalanine production in these mutants was inhibited by L-tyrosine and stimulated by L-tryptophan.

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dl-Phyllostine and dl-epoxydon, phytotoxic metabolites of Phyllosticta sp., have been synthesized by regiospecific procedure.

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Synthesis of erythroidine skeleton, containing β, γ-unsaturated-δ-lactone system derived from erythrinane skeleton was described.

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A stereoselective synthesis of methyl trans, trans-farnesoate (I) from isoprenyl bromide and geraniol is described. dl-C₁₇-Cecropia juvenile hormone (IV) was synthesized from it in a stereoselective manner.

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Hydrophobicity and amounts of polar groups on the cell surface were determined by the use of fluorescent probe, colloid titration, and acid titration. Surface hydrophobicity of hydrocarbon-grown cells was about 7 times greater than that of glucose-grown cells of the same strain, while amounts of polar groups did not differ so much. Adsorption of cells to air bubbles was maximum at pH 3. Langmuir's adsorption isotherm held for the adsorption. Affinity to air bubbles of hydrocarbon-grown cells was 1.8 times greater than that of glucose grown cells, while the theoretical maximum amounts of cells to adsorb per unit area of bubbles were equal for the both cells.

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Corynebacterium glutamicum mutants carrying both auxotrophy and histidine analogresistance were derived by a mutagenic treatment, and their histidine productivity was compared with that of a triazolealanine (TRA)-resistant histidine producer, C. glutamicum KY-10260. As a result, a leucine auxotrophic TRA-resistant mutant, Rα-88 was selected out of 164 auxotrophic derivatives of KY-10260. It produced histidine at a distinctly higher concentration than the parent strain under every condition tested. The concentration reached 11mg/ml or 5.8% (w/w) of the initial sugar. Addition of an excessive amount of leucine to the medium inhibited the histidine production together with the by-production of valine by this mutant. Thiazolealanine-resistant mutants derived from a tyrosine auxotroph, a phenylalanine auxotroph and a tryptophan auxotroph gave the same or lower production in comparison with KY-10260.

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A riboflavin α-glucoside-synthesizing enzyme from the acetone powder of pig liver was purified by a procedure including fractionation with ammonium sulfate, heat treatment, fractionation with acetone, gel filtration on a Sephadex G-150 column, calcium phosphate gel treatment, and isoelectric focusing. A final enzyme preparation was homogeneous on polyacrylamide disc gel electrophoresis and in the ultracentrifuge. The enzyme had a sedimentation coefficient of 9.90 S and an isoelectric point of pH 3.7. The enzyme had a pH opti-mum at 6.0 with maltose as substrate. The enzyme catalyzed the hydrolysis of diverse kinds of α-glucosidic substrates, and the transfer of α-glucosyl residue from these substrates to riboflavin. The Km value for maltose was 1.20×10^-3M. The enzyme hydrolyzed phenyl α-maltoside to glucose and phenyl α-glucoside. Amylose was almost completely hydrolyzed to glucose by the enzyme. Maltotriose was obtained as the main transfer product after the treatment of maltose with the enzyme. The enzyme also catalyzed the transfer of α-glucosyl residue from maltose to pyridoxine, esculin, rutin, and adenosine. It was recognized that a single enzyme catalyzed not only the hydrolysis of maltose and α-glucosidic substrates but also the transfer of the α-glucosyl residue of these substrates to suitable acceptors.

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A novel metabolite from Gibberella fujikuroi was isolated and its structure was elucidated as 4β, 7β-dihydroxy-18-norkaurenotide (I).

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