Variation in morphology and penicillin run down of Pen. chrysogenum Q 176 and the pigmentless sultant strain during twelve generations of successive subcultures on eleven different media as shown in Table 1, with special reference to the influence of C. N. ratios of media, nutrient rich and poor media on the above variation of these molds were studied.
Mutation rate and penicillin run down at each generation are shown in Table 1 and 2.
In the case of the medium containing no organic nitrogen, majority of the spores of the culture turned to the flat type mutant, while in the case of medium rich in peptone, the heavy flat type mutant, which were both described in the Part 5 of this report, predominated.
No remarkable morphological variation and penicillin run down were recognized in M_OYER medium and modified M_OYER medium even after twelve successive subcultures.
Then the authors tried to propose some genetical explanations on the phenomenon of the variations or occurence of many mutants derived from Pen. chrysrgenum Q176 and on the various experimental data of several aspects of variation of this mold which have been obtained in the previous sixteen reports.
In this report, a method of developing mutants from the various mutants already obtained by further irradiation of ultraviolet ray upon them and a method of repeating irradiation on the newly induced mutants were adopted.
And by this method a sheme of developmental lines of mutants from Q176 was drawn as in Fig. 3.
It was presumed, from the analogy of the. gene mutation of Neurospora by B_EADLE et at., that mutations in spore formation, penicillus formation, coloration of spore and the yellow pigments synthesis etc. were all considered as one step of gene mutation.
Thus some new knowledges was added to the explanation of the variation of Penicillium which propagates by non sexual way and the reports conerning this point have been rare so far.

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The colour of shoyu (soy-sauce) was studied by H_ARDY's self-recording spectrophotometer in the region of visible spectrum ranging from 400 mu to 740 mn and represented by the C. I. E. -method. The following results were obtained:
1. The colour of shoyu represented by the C. I. E. -method is dominant wave lengh 590_??_620mμ, excitation purity 86_??_88%, and luminous transmittance 0.14_??_0.17.
2. Shõyu has stronger absorption in the shorter wave length and weak one in the longer wave length in the visible spectrum. The form of its trans mittance curve remains unchanged even when the process of its preparation is varied.
3. The degree of colour deepness depends upon the kind'of shoyu. If compared on the basis of total nitrogen content reduced to 1%, colour deepness may be arranged in the following order.: Aminosan shoyu (prepared by low-temperature decomposition)>Shinshiki shõyu>Kiage-shõyu >Aminosan-shõyu (prepared by high-temperature decomposition.)
4. By treating with active charcoal, difference in colour is found between Kiage-shõyu and Aminosan-shoyu; namely, Kiage-shoyu shows characteristic absorption effect in the longer wave length.

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The experimental results are as follows.
1. The colour change of soy sauce during storage is not due to enzymatic reaction, the action of air being the most effective and the actions of temperature and light unremarkable. This shows that air-oxidation is the principal cause.
2. When the colour of soy sauce is deepened by aeration, the form of transmittance curve differs no remarkable change.

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1. The author concludes that the cell-free extract from ground dried cells of Bact. succinicurn S_AKAGUCRI et T_ADA has the activity of glycerol-oxidation.
2. The extract contains some enzymes and, especially, is able to oxidize acids of the citric acid cycle.
3. ATP accelerates the glycerol-oxidation.
4. α-glycero-phosphoric acid is found in a resultant medium from enzymatic glycerol-oxi-dation by paper partition chromatograpny.

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1) An attempt has been made to use a new apparatus for a submerged fermentation ror the production of the antibiotic substance which inhibits the growth of Lthe pellicle forming, yeast of “shoyu” (Zygosaccharomyces).
2) Some good results have been obtained by an experiment on the productiou of the antibiotic substance-with this small apparatus, which has a glass cylindrical side-wall and a special agitating rotation-tube for blowing the air into broth.
3) The fundamental datum on the large tank culture of the antibiotic snbstance can be taken by using this apparatus.

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Authors investigated the continuous molasses7fermentation based on the growth and fermentation curves by two vessels system. Various conditions affected to the growth and fermentation, and continuous fermentation cycle times in primary fermentor was examined.
To operate the continuous fermentation according to A_DAMS' theory, the following theorerticalrelation between the growth and fermentation curves must be exist in each time.
Total Yeast Cells/ Yeast Cells Produced per hr.=Total Sugar Fermetnted/Sugar Fermented per hr.
When the amount of inoculated yeasts are nht negligible to the produced yeasts, the con-tinuous operation cau not be attained by toe cycle time calculated from the analysis of the .growth curve. In such case, the steady state of fermentation can be maintained readily, if the cycle time which is calculated from the amouut of the produced yeasts after renducing the equal quantity of the yeasts corresponding to the inoculum size from the fermenting mash.

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1. By using the method of paper-chromatography, the organic acids production by molds in ghicose medium were extensively studied. Among 14 strains of Asp. oryzae, three strains were found that they produced α-ketoglutaric acid along with other acids such as kojic, citric, malic, lactic, succinic and fumaric acids by surface culture. These acids except α-keto-lutarate were found by one of the authors⁽¹⁾ to be the products fo Asp. oryzae.
2. Two strains which did not produce α-ketoglutaric acid by surface culture formed this acid under a submerged state.
3. Two strains of Rhizopus belonging to fumaric acid former produced α-ketoglutaric acid by surface culture. The fact was confirmed not only by the method of paper-chromato-graphy, but also by obtaining the pure material from the fermentation medium as 2, 4-dini-trophenylhydrazone.
4. Recently W_ALKER et al.⁽²⁾ reported the presence of α-ketoglutaric acid in the fermentation medium of glucose by Asp. niger. As we found this acid not only in the medium of Asp. oryzae but also by Rhizopus species, so it may be concluded that α-ketoglutaric acid is a normal product of various molds in glucose medium.

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When various quantities of KOH solution are added to the Cr-liquors (Cr₂(SO₄)₃, K[Cr(C₂O₄)₂], K₃[Cr(C₂O₄)₃], K[Cr(C₂O₄)₂_]+Cr₂(SO₄)₃, K₃[Cr(C₂O₄)₃]+Cr₂(SO₄)₃) a gradual change occurred on the reading of photocolorimeter.
When these changes are plotted on a graph, the lines thus formed may be divided to two hypothetic straight lines; one line may be due to colour change and the other to the gel formation.
As the results of the studies on these absorption spectra, it is presumed that every chrome solution would have the same colloidal condition at the intersecting point of the above two straight lines.
After the hide powder was, mixed to these chromes which may be in the same state as mentioned above, the quantitative determination of the chrome fixed to the hide powder was carried out. It is revealed that every chrome is fixed to the hide almost with the same quantity after 20 hours, but the mitxtures of anionic- and cationic-Cr are more promoted in the velocity of fixation to the hide than the singly used cationic.

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The end group assay was carried out with periodate oxidation method on several starchtype polysaccharides, and average number of glucose residue per non-reducing end group was calculated from the amount of formic acid liberated after 25 hours of oxidation by 0.037 M sodium metaperiodate at 0° The following values of average chain length were obtaind: soluble potato starch 29.5, potato amylopectin 26.3, potato amylose 480, soluble glutinous-ricestarch 18, and polysaccharide produced from glutinousrice starch by the action of isoamy lase 100±5. The value for the product of isoamylase approximately coincided with values of average polymerisation Ldegree, formerly estimated with viscosimetryor osmometry.
From the above result the authors have confirmed KOBAYASHI's theory that the isoamylase produces amylose-type polysaccharide from amylopectin-type polysaccharide as previously, reported⁽¹⁾⁽²⁾.

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The distribution of thiamine in the domestic wheat, and covered and naked barleyes at different stages of maturity has been studied by the photographic method of thiochrome fluorescence, and the moisture contents, the weight of 1000 kernels and the thiamine content have been determined
It has been studied that there is no significant difference among wheat and covered and naked barleyes as to the following items
(1) The fluorescence of thiamine is remarkable in the ovary wall surrounding the kernel at an early stage.
(2) The thiamine is not yet concentrated in embryo at an early stage of development othf new embryo.
(3) When the kernels reach the yellow stage of maturation, the quantity of thiamine in -the embryo increases steadily, then the ovary wall becomes pericarp and has, more concentrated thiamine.
(4) From the early stage of development, the distribution of thiamine is more heavily concentrated in the alleurone layer but is comparatively less in the endosperm.
It is supposed that thiamine of wheat kernel removes through the vascular bundle from other tissue of plant body and pass the placenta to the growing kernel.

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The distribution of thiamine in the rice at different stages of maturity has been studied by the photographic method of thiochrome fluorescence, and the moisture contents, the weight of 1000 kernels, and the thiamine contents have been measured.
The thiamine in rice has been concentrated rapidly during rapid development of tissue. At the extreme early stage the thiamine fluorescence has appeared in pale with a comparatively heavy concentration. Until 5 days after blossoming the thiamine fluorescence has been distributed mainly in the ovary wall surrounding the kernel. In 10 days it has more heavily concentrated in embryo. In 15 days the ovary wall has become pericarp and had a lot of thiamine, thereafter the distribution of thiamine fluorescence has had no change to the last maturation period.
The distribution of thiamine fluorescence of rice during the maturation has been different from wheat or barley. In case of rice the distribution has been completed within a half period as long as in case of wheat or barley.
The rice kernels at the late maturation period has been dried naturally or by dryer, but there has been little difference as to their distribution in the kernels.

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(1) A large amount of ergot alkaloids is produced within appreciable short times in surface replacement cultures.
(2) The ergot alkaloids are not produced when the growth of the fungus does not take place even in surface replacement cultures.
(3) The alkaloid production in surface replacement cultures is not so much affected by the natures of C- or N-sources as in the ordinary surface cultures, as long as the age of mycelium is not so young or so old and the depth of the culture liquid is not so high.
4) The surface mycelium of ergot fungus continues to produce the alkaloids with undi-minished vigor through several successive replacement of culture liquids. The alkaloid production in this semi-continuous process fundamentally depends upon the surface mycelium first obtained.
(5) The ergot alkaloid production in surface replacement cultures is much affected by the age of mycelium or by the depth of the replacement liquid.

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(1) In order to produce the ergot alkaloids by surface replacement cultures, it is reasonable to use such a culture medium which has rather high C/N ratio due to rather, low, nitrogen concentration.
(2) In order to produce the ergot alkaloids by surface replacement cultures, it is rea-sonable to carry out the chltivation at relatively low temperature. In general about 24° is, preferable.
(3) The alkaloid production in surface replacement ent cultures is not so much affected by-the initial pH of the culture media as in ordinary surface cultures. The alkaloid production: in the former is not, affected appreciably at the pH values between 5.0 and 6.0.
(4) The alkaloid production in surface replacement cultures is also enhanced by various, growth inhibitory substances in the culture media. But in replacement cultures, unlike in, ordinary surface cultures, the said effect can be seen for the first time when the concentr-ation of the substances is fairly increased.
(5) The mycelium of ergot fungus grown on the surface of a suitable, liquid medium, produces the alkaloids rather excellently even in submerged cultures.

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When the acetylesterase (AcE) fraction of Sclerolinia fungus extract was added together with blood cholinesterase (ChE) to the solution of acetylcholine (Ach) and the rate of hydrolysis of Ach was measured by Warburg manometric method, it was found that the distin-ct inhibitory action upon ChE appeared. Heat-inactivated AcE fraction had also the same effect. It was thought that the inhibitory action of AcE upon ChE was probably depedent on a peculiar mechanism of AcE action. AcE seemed to restrict the substrate to some ex-tent by formation of inactive enzyme-substrate complex. AcE was inactivated by dialysis for a long time and lost the inhibitory action upon ChE, but the dialyzed solution was reactivated by adding the heat-inactivated enzyme solution or the outer solution of dialysis. It was supposed that the essential group, coenzyme of AcE required for combining with substrate remains after heating and accordingly heat-inactivated enzyme solution has also the inhibitory action upon ChE owing to the ability to form inactive complex with substrate. Uterus-contracting constituent of Sclrotinia fungus extract as already reported is thought to be closely related with the chemical characteristics of AcE and the mechanisms of.its physiological action may he dependent on the inhibitory action updn ChE.

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1. The influence of α, α'-dipyridyl on the citrate formation from glucose by the fungus mat of Asp. niger was studied. The reagent had been found by the authors to inhibit the anaerobic decomposition of citrate by Bact. succinicum.
The remarkable increase of citrate formation occured in the glucose medium containing M/1000 α, α'-dipyridyl. This effect was always observed even in the purified glucose medium containing no ferrous ion, so it was concluded that this effect was not due to the deprivation of ferrous ion from the medium by the reagent.
2. Aerobic decomposition of citrate by two strains of Asp. niger, one the citrate former (S-292-A ) and the other without the ability of citrate production (No. 26), was greatly inhibited by M/1000 α, α'-dipyridyl. This inhibition of citrate decomposition was presumed to be the cause of the promotion of citrate formation from glucose by the reagent. But in the case of No. 26, the addition of dipyridyl did not result the accumulation of citrate.
3. The aerobic decomposition of acetate by these strains was also inhibitid by α. α'-dipy ridyl. The degree of inhibition of acetate oxidation by the reagent. was almost equal to that of citrate. So it was concluded that acetate was oxidiied by a way of tricarboxylic acid cycle.
It was not clear enough that, what reaction of tricarboxylic acid cycle was inhibited by dipyridyl, but it was demonstrated by paper-chromatography that aconitase was not inhibited.

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The phosphorus turnover of various fractions of yeast (Sacch. sake and Zygosacch. soja) and fungi (Asp. oryzae A) have been studied with the aid of P3 as a tracer.
Glucose promoted the phosphorus turnover of yeast in general. This effect was tested under various different experimental conditions and always recognized. (Table 1_??_6). But when the amount of glucose added was small, this effect was not apparent (Table 2).
The phosphorus turnover of yeast was generally higher under the aerobic conditions than the anaerobic (Table 5).
The rate of turnover was different according to the species tested, but the general features were identical (Table 2 and 5). Small amount of ammonium sulfate did not influence the phosphorus turnover, but, if the concentration was higher it promoted the turnover slightly. Ammonium tartarate, on the other hand, promoted more remarkably the phosphorus turnover of acid insoluble fractions of yeast (Table 6).
The effect of glucose and nitrogen compounds on the phosphorus turnover of fungi were

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Previously, the author reported that the mass production of Ca-2keto-L-gulonate was successful from Ca-L-idonate (in practice an equimolecular mixture of Ca-L-idonate and Ca-D-gluconate, abbreviated as Ca-I-G had been used) by the freshly isolated Ps. fluorescens sp., and also reported, that vitamin C was prepared from the Na-salt of Ca-2-keto-L-gulo. nate thus obtained.
Repeating a further investigation, -the author succeeded in obtaining only pure 2--keto-L-gulonic acid in a crystalline state from the broth. The results are summarized as follows:
Repeating a further investigation the author succeeded in obtaining only pure 2--keto-L-gulonic acid in a crystalline state from the broth. The results are summarized as follows:
1) In such a case as; bouillon agar slant → Ca-I-G agar slant → Ca-I-G shaking culture, initial pH 6.1→elapsed from pH 7.0 to pH 5.8 → final broth (15 days of total incubation period) ; the max. yield in a crystalline state was 42.2% to Ca-I-G.
2) In such a case as; 7 times repeating of inoculation of Ca-I-G agar slant → Ca-I-G shaking culture, initial pH 6.1 4 days →elasped pH 7.0 4 days→elasped controlled to pH5.8 6 days→elasped final broth (14 days of total incubation period); the max. yield was 45.2%.
3) In the case as ; cells incubated 50 days on Ca-I-G agar slant → Ca-I-G agar slant → Ca-I-G shaking culture, initial pH 6.1 4 days →elasped PH 7.0 2days→elasped controlled to pH 5.8 7days→elasped final broth (11 days of total incubation period), the max. yield in a crystalline state rised up to 52.8% (purity:94%). The author found that it was more favorable to control the pH value to 5.8 than to 5.5 in the above described manipulation, in order to obtain the less deviated yields.
4) By using the strain, obtained by the methods illustraed in column (3), the successful, yields of 2-keto-hexonic acid was obtained. In this case, the maximum yields of 2-keto-L-gulonic acid in a crystalline state through this investigation was given as well. The further decomposition of 2-keto-ic gulonic acid by this strain was hardly recognized.

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The environmental effects upon anastomosis of Koji-molds have been studied by means of hanging drop culture and mixed point-inoculum plate culture, using the color mutants of Asp. sojae. (1 and 2)
1) The lower concentrations, i. e. about 1/10 of ordinary concentrations, were generally-favorable upon various natural media, such as Koji-, Malt-, Rice germ-, and Yeast-extracts. The frequency of anastomosis seems to be accerelated by poor nutritional conditions.
2) Upon the synthetic media (Czapek solution), from 1/2 to 1/20 of the original concent-rations gave good results. Any significant effects could not be recognized through changi-ng the concentrations of KH₂PO₄, KCl, and MgSO₄ in Czapek solution.
3) The optimum concentrations of NaCl, glucose, and lactose, in distilled water were 0. 1 mol., 0.2 mol. and 0.1_??_0.5mol. respectively. Higher or lower osmotic pressures have been. found to be less favorable.
4) The higher C/N ratio generally favours anastomosis especially when the contents of N-source in the media are low to definite extent.
5) The range of the optimal pH zone was rather narrow, i. e. from 5.4 to 6.0. The ability of anastomosis decreased rapidly at the lower, and slowly at the higher side of the optimal zone and was completely inhibited at pH 4 and pH 8.8.
6) The age of the inoculum cultures upon Koji-extract agar at 30° does not give any significant influence upon anastomosis in the extent of from one week to three months. The-inoculum of older cultures, for instance, 7 months, seems to decrease the frequency.
7) The optimum temperature for anastomisis lies in between 25° and 30° which are som-what lower for the optimum growth of the molds.

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1. The tannin, water-soluble matter, and crude fiber contents of two varieties, one for green tea and the other for black tea, produced in the Kagoshima, Shizuoka, and Saitama Prefectures were determined and the local variation was analyzed statistically.
2. The tannin content was higher in the summer crop than in the spring one, and also was it in the product of Kagoshima than in the product of Saitama. And the leaf of C-8 (black tea variety) contained more tannin than Yabukita (for green tea variety).
3. As for the soluble matter, only the varietal difference was significant which, showed C-8 to be richer than Yabukita.
4. The ratio of tannin/soluble matter was larger in the product of Kagoshima than in the other two. Especially in the second season, the ratio was larger as the latitude compared to the producing district was lowered, so it was recognized that the black tea production was advantageous in southern district.
5. The ratios of soluble-N/soluble matter and amino-N fraction/soluble matter were larger in the spring crop than in the summer one; they showed the reverse tendency conpared ot the tannin content. The ratio of the amino-N fraction, which was considered the most important element of the taste of green tea, was larger in Yabukita on an average, but the seasonal difference was remarkable, and this fact could explain the fluctuation of the qualities of tea made from Yabukita variety.
6. The crude fiber content was lower in the product of Saitama than in the other two, and also was it in the spring crop than in the summer one. This tendency was reverse for the shoot length of the samples.

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1. The local difference of the ash and its alkalinity of the tea leaf was examined, sampled: from the varieties of Yabukita (for green tea) and C-8 (for black tea), grown in the Kagoshima, Shizuoka, and Saitama Prefectures, during the three years from 1949 to 1951.
2. Of all the constituents, including those in already reported papers, the ash and its. alkalinity indicated the most remarkable local difference. The tea leaf produced in the Ka-r goshimna Prefecture containd much less ash and alkalinity than that of Saitama.
3. As it was considered that the local difference of ash and alkalinity might be due to the soil components, the soils of the tea gardens from which the samples had been collected were analyzed, and P₂O₅ and MnO contents showed significant local difference which proved that the Saitama soil was abundant in these ingredients than the soils of other two places.
4. However, the relation between soil constiuents and leaf ash or alkalinity was not distinct, except that between soil alumina and leaf alkalinity whose correlation coefficient was calculated as +0.96.
5. Annual and seasonal differences were considered to be greatly affected by climatic conditions, but any decisive relation could not be obtained as far as this experiment is con-cerned.

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A new method has been devised for the synthesis of phthaloyl dipeptide esters. Phthylimidoacyl chloride and amino acid ester readily reacted to give corresponding phthaloylpeptide ester, Removal of the phthaloyl group after saponification with dilute alkali lead to free dipeptide.
Procedures are given for the synthesis in a high yield of glycyl-L-leucine, and eight phtha-loyl dipeptide ester containing glycine, alanine and leucne.

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We have synthesized sixteen kinds of mercaptals and two kinds of mercaptols. Eight of them are unknown substances. Odor of them are iilustrated in Table 1

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