• Applied Biological Chemistry
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• v.4
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• pp.33-42
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• 1963

1. Two fermentation methods have been compared for the production of Takju (Korean native Sake). One method (S1) uses 'Nuruk' (Korean mold wheat) and the other method (S2) uses 'Nuruk' and mold rice. The chemical analysis of source material, also, have been made in order the check the quantitative changes during the fermentation. The results are summarized below: a. The rate of fermentation was slower for the S2 method than for the S1 method. However, the residual sugar in the S2 method was smaller and the amount of alcohol produced the same as with the S1 method. This was true in spite of the fact that the total initial sugar content for the S2 method was 10 percent below that of S1. b. With both the S1 and S2 methods, 80 percent of the total alcohol production was obtained with in 3 to 4 days. c. The pH value of the base material at the beginning of fermentation was markedly different between the two methods I.e.6.0 to 6.2 for S1 and 4.8 to 5.2 for S2. However, after one or two day the pH of both materials was about the same 4.0 to 4.2 and remained at this value unit the fermentation was complete. 2. Organic acids and Sugars in the source materials have been detected by the paper partition chromatography(p.p.c.) method and the followings are obtained. a. Important Organic acids are Fumalic, malic Succinic, Citric, Acetic aicds in polished rice and Fumalic, Succinic, Acetic, Citric, Malic and Oxalic acids 'Nuruk'. The same kinds of acids as in the rice are found in mold rice, However, amount of Citrice acid is markedly increased in mold rice. b. The important Sugar, Glucose, Fructose and Raffinose in polished rice, Gulcose, Fructo, se, Xylose and one which supposed to be Kojibjose in 'Nuruk'. Glucose and those supposed to be Isomaltose, Kojibiose and Sakebiose and found in rice mold, however, no Sucrose, which was exist in polished rice, was found. 3. The important Organic acids found in fermenting mash using the p.p.c. method were Lactic, Succinic and Acetic. Citric acid identified early in the fermentation, S2 method. remained throughout. However, with the S1 method Cirtic acid was detected only during the late stage of fermentation. Sugars not found in the original materials were two which supposed to be Isomaltotriose and Pentose. Maltose found at the beginning of the fermentation disappeared within one day, Isomaltose was detected throughout the period of fermentation. 4. The Somogyi method which was employed to determine the quantitative changes of sugars in the orginal meterial and mach, showed that polished rice containes in order and in largest amounts Sucrose, Glucose, Raffinose and Fructose. 'Nuruk' contained almost equal quantities of Glucose and Fructose. However, the Glucose content of the mold rice exceeded that originally in the polished rice by 25 or 30 times. Only a small quantity of free sugars was found in the mash at the end of the fermentation.

• Korean Journal of Agricultural Science
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• v.9 no.1
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• pp.284-302
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• 1982

Development of protein resources as food has been a big issue especially in Southeast Asia region, and intake of protein is also insufficient in Korea. To cope with this shortage of protein resources and its improvement together with increased production of high nutritive animal protein, studies were carried out on feeding of Korean native goats. In the experiments were made absorption of carbohydrate and volatile fatty acid in miniature rumen, and absorption of amino acid in rumen as in vivo were conducted as part of studies on nutritional absorption in rumen. Those nutritional for improvement of feeding and management as described above are summarized as following. 1. According to the result of test on the nutritional absorption of native goat by means of miniature rumen method, absorption ratio of VFA measured at 0.5, 1 and 2 hours after injection of nutrition showed propionic acid 70-86%, acetic acid 74-87%, and lactic acid 76-89%. In the absorption of organic substances, ethyl alcohol of 0.5% showed 29-87% and lactic acid of 0.1M showed 12-27% of absorption ratio. 2. Result of absorption measurement in rumen from L-type free amino acid injection in the content of rumen vein showed lower rate at menthionine-free group compared to whole-egg amino acid injection in the content of rumen vein showed lower rate at methioine-free group compared to whole-egg amino acid group, and high absorption ratio was observed at methionine 3 times group and urea added group. Deficiency of methionine caused no change of the content in mucous membranes. 3. Absorption of amino acid in rumen muscular layer showed equal tendency as in the mucous membrane without exerting any influence of methionine deficiency. At the methionine3-times group, content of methionine and glutamine were increased by 14.7 and 4.4 times as compared to whole-egg amino acid group, an absorption ratio of glutamine, proline and valine were increased at urea added group. 4. In general, concentration of amino acid in rumen vein plasma was lower than in rumen mucous membrane and muscular layer. Absorption ratio of amino acid is decreased due to methionine deficiency, and tripling of methionine or urea adding caused increment of amino acid. Absorption pattern is thus varied depending on the composition of amino acid. 5. At the urea added group, content of ammonia-N, amino-N and urea were increased in rumen muscular layer. As the inside of goat's rumen was unable to clean thoroughly, investigation was made on remaining bacteria, however, variation of ammonia-N was affected by these bacterial content. 6. Variation in rumen structure by differential absorption of amino acid was observed by general microscope and fluorescent microscope. According to the result of observation in the methionine 3 times group, single cylinder epithelium of mucous membrane showed rather thin, and it was thick at urea added group though no significant differences existed among test groups in submucous membrane and muscular layer.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.507-519
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• 1999

The ultimate objective of periodontal treatment is to stop disease progression and to regenerate destroyed periodontal tissues and thereby regain normal function. Growth factors are naturally found polypetides which stimulate many cellular activities pertaining to wound healing by acting as signal molecule in controlling cell movement, proliferation, and matrix production. Platelet derived growth factor (PDGF) is 28,000-35,000 Da molecular weight dimeric protein with 2 long positively charged polypeptide chains connected by sulfide bonds. The purpose of this study is to evaluate histologically the initial guided tissue regeneration in a periodontal defect f a beagle dog treated with a biodegradable membrane formed with polylactic acid (poly-L-lactic acid) and polyglycolic acid loaded with 200ng/$cm^2$ platelet derived growth factor. 2 beagle dogs were used in he experiment. $5mm{\times}6mm$ alveolar bone defect was formed in upper and lower canines and third premolars and a reference notch was placed. PDGF-BB non-containing membrane was used as control. Each defect was randomly assigned to the test roup or the control group. The dogs were sacrificed 3 weeks after membrane placement. Toluidine blue and multiple staining was done for histological analysis. In the 3 week specimen in the control group, no new one formation could be seen. Small amount f bone resorption below the notch could be seen. In the notch, loose connective tissue with infiltration of inflammatory cells could be seen. Also thin discontinuous new cementum could be seen and the membrane still retained its structure. Where PDGF-BB containing membrane was used, new bone formation could be seen in the notch at weeks and also continuous thin cementum could be seen. PDL cells were observed between new bone and new cementum and some were attached to bone and cementum. These results suggest that new bone and cementum formation seen when PDGF-BB loaded membrane was used was due to inhibition of downgrowth of epithelial cells and also due to continuous release of the growth factor. Further study on the resorption characteristics of the membrane nd the release characteristics of the PDGF-BB is necessary. Also, development of a membrane easier to use clinically is necessary.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.6
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• pp.808-813
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• 2004

This study was conducted to evaluate the effects of adding glucose (G), sorbic acid (S), pre-fermented juice of epiphytic lactic acid bacteria (FJLB) and their combinations on the fermentation qualities and residual mono-and di-saccharides compositions of guineagrass silage. The additives used in this experiment were 1% glucose, 0.1% sorbic acid and FJLB at a theoretical application rate of 9.0${\times}$105 CFU $g^{-1}$ on the fresh weight basis of guineagrass, respectively. There was a total of eight treatments in this experiment: (1) C (without additives), (2) FJLB, (3) S, (4) G, (5) FJLB+S, (6) FJLB+G, (7) S+G, (8) FJLB+S+G. After 30 days of storage, the silos were opened for chemical analyses. Based on the results, all additives were efficient in improving the fermentation quality of guineagrass silage. This was well indicated by significantly (p<0.05) lower pH and BA content and significantly (p<0.05) higher LA content in the treated silages except for the FJLB than in the C. However, there was only a slight increase in LA for the FJLB as compared with the C, which might be due to the low WSC content of the original guineagrass (34.4 g $kg^{-1}$). When the FJLB+S and FJLB+G were added, there were significant (p<0.05) decreases in pH and significant (p<0.05) increases in LA as compared with the FJLB alone. This indicated that the G, S and FJLB were of synergestic effects on the silage fermentation quality. The G combination treatments including the G alone showed large improvements in the fermentation quality as compared with the treatments without the G. This suggested that adding fermentable substrates (G) to plant materials such as guineagrass, which contain low WSC, intermediate population of epiphytic LAB, CP and DM content, is more important and efficient for improving the fermentation quality of silages than adding a number of species of domestic LAB (FJLB) and aerobic bacteria inhibitor (S).

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.6
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• pp.1050-1057
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• 1997

Microbial counts, enzyme activities and taste components of traditional kochujangs prepared with the powders from 4 different varieties of red pepper, were investigated during 90 days fermentation for the industrial production of traditional kochujang. The viable cell counts of anaerobic bacteria in kochujangs did not change remarkably during fermentation, however, aerobic bacterial counts showed a rapid increase up to 90 days of aging. The yeasts in all kochujang samples increased until 60 days of aging and than decreased. After 90 days of aging, the count of aerobic bacteria in Kumtop kochujang was higher than those of others. The activities of liquefying amylase decreased during the aging, but those of saccharogenic amylase increased at 60 days of aging. The activities of neutral protease were higher than those of acidic protease, and increased during the middle and last period of aging. The major free sugars of kochujang were maltose and glucose, and their contents were higher in Hongkwang kochujang. The major organic acids of kochujang were succinic, formic and citric acid, followed by lactic acid. Succinic acid content in kochujang decreased during fermentation, whereas formic and citirc acids were increased. The major free amino acids were serine, proline, glutamic acid, aspartic acid and alanine. Kumtop kochujang contained the highest amount of total free amino acids. Among the nucleotides and related components in kochujang, cytidine-5-monophosphate was the most abundant component at the begining of aging period, while hypoxanthine increased remarkably during fermentation. Hanwang kochujang was higher in the content of nucleotides than others. Capsaicin contents of kochujang decreased during aging and those of Jangter kochujang was higher than that of others. Sensory evaluation showed that Jangter kochujang was significantly better than Kumtop kochujang in overall acceptability, but there were no appreciable differences in color and flavor.

• Food Science and Preservation
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• v.10 no.3
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• pp.339-346
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• 2003

In order to improve some problems such as contamination of undesirable mold, mycotoxin production and excessive drying on the surface of traditional meju, we developed a capsule type-meju(CM) coated with steamed black bean and Aspergillus oryzae - rice koji(0.3％, w/w) mixture to surface of traditional meju and fermented at 25$^{\circ}C$ for 14 days under 80％ relative humidity. Contamination of undesirable mold on the surface of CM was not detected within 2 weeks and some genus Penicillium molds on control meju without koji were found naturally after 12 days of fermentation. The moisture content of meju was showed to be in the range of 34.7 - 29.4％ being 32.7％(w/w) of mean value. Titratable acidities in CMs prepared with black bean(BCM) and soybean(SCM) were much higher than that in control meju, and BCM was similar to SCM. Free sugar content in BCM(123.98 mg％) was 10 times and 2.1 times higher than that in control meju(15.02 mg％) and SCM(59.85 mg％), respectively. Amino type nitrogen content in control meju was 147.00 mg％ and its content in BCM(255.50 mg％) was 1.37 times higher than that in SCM(187.25 mg％). Total organic acid content in BCM(95.98 mg％) and SCM(1l9.98 mg％) were much higher than that in control meju(26.44 mg％), and then lactic and malic acid contents were markedly changed according to capsulation of meju. Lightness value(L) of Hunter color index was much higher in BCM than in SCM. Fatty acid composition of CM was not different as compared to control meju. Total free amino acid content in BCM(1039.70 mg％) was 4.4 times and 2.4 times higher than that in control meju(236.45 mg％) and SCM(556.07 mg％), respectively.

• Journal of Animal Science and Technology
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• v.50 no.1
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• pp.89-98
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• 2008

This experiment was carried out to investigate the combination feeding of β-glucan and multi-species probiotics on growth performance, various lipid concentrations of serum, antibody production and cecal microbial profiles in broiler chicks. A total of six hundred 1-d-old male broiler chicks were divided into five groups, placed into four pens per group(30 birds per pen) and fed one of five non-medicated experimental diets(T1, 0.15% multi-species probiotics; T2, 0.1% β-glucan+0.15% multi-species probiotics; T3, 0.3% multi-species probiotics; T4, 0.1% β-glucan+0.3% probiotics or devoid them as control) for 5 wk. There was no significant difference in feed intake among the groups. The average weight gains and FCR in groups fed diet containing 0.3% probiotics were significantly improved(p<0.05) than control in finisher period(22-35d). The concentration of serum cholesterol ester in groups fed 0.3% probiotics were significantly lowered(p<0.05) as compared to that of the control. Relative weights of liver, spleen, bursa of Fabricius, breast and leg were not influenced by the dietary treatments. The average ND or IB antibody titers in groups fed diets containing β-glucan and probiotics were tended to be increased, but not significantly. The number of cecal lactic acid bacteria was significantly increased(p<0.05) by the dietary β- glucan and probiotics. These results indicated that dietary β-glucan and probiotics exerted a growth- promoting and immune-enhancing effects on broiler chicks. In addition, yeast derived β-glucan, and multi-species probiotics modulated the profiles of cecal microflora, reflecting potential alternative substances to replace antibiotics for feeding broiler.

• Journal of The Korean Society of Grassland and Forage Science
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• v.36 no.1
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• pp.7-14
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• 2016

The objective of this study was to determine the effect of bacterial inoculation (Lactobacillus plantarum or combo inoculant mixed with Lactobacillus plantarum and Lactobacillus buchneri) and addition of fibrolytic enzyme on chemical compositions and fermentation characteristics of whole crop barley (WCB) and triticale (TRT) silage, their ruminal in vitro fermentation, and digestibility. In TRT silage, enzyme addition significantly (p<0.01) decreased NDF content compared to no enzyme addition treatment. Organic acids such as lactate and acetate contents in WCB and TRT silages were significantly (p<0.01) higher compared to those in the control. Particularly, lactate content was the highest in L. plantarum treatment. Fibrolytic enzyme treatment on both silages had relatively higher lactic acid bacteria content, while mold content was lower in both treatments compared to that in the control. In vitro dry matter digestibility was generally improved in WCB silages. It was higher (p<0.01) in TRT with mixed treatment of L. plantarum, L. buchneri, and enzyme compared to others. In vitro ruminal acetate production was relatively higher in treatments with both enzyme and inoculant additions compared to that in the control. Therefore, the quality of silage and rumen fermentation could be improved by inoculants (L. plantarum and L. buchneri) regardless whether whole crop barley (WCB) or triticale (TRT) silage was used. Although it was found that fibrolytic enzyme addition to both silages had various quality and rumen fermentation values, further study is needed.

• Journal of Microbiology and Biotechnology
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• v.32 no.5
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• pp.630-637
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• 2022

The objective of this study was to optimize industrial-grade media for improving the biomass production of Weissella cibaria JW15 (JW15) using a statistical approach. Eleven variables comprising three carbon sources (glucose, fructose, and sucrose), three nitrogen sources (protease peptone, yeast extract, and soy peptone), and five mineral sources (K₂HPO₄, potassium citrate, ⳑ-cysteine phosphate, MgSO₄, and MnSO₄) were screened by using the Plackett-Burman design. Consequently, glucose, sucrose, and soy peptone were used as significant variables in response surface methodology (RSM). The composition of the optimal medium (OM) was 22.35 g/l glucose, 15.57 g/l sucrose, and 10.05 g/l soy peptone, 2.0 g/l K₂HPO₄, 5.0 g/l sodium acetate, 0.1 g/l MgSO₄·7H₂O, 0.05 g/l MnSO₄·H₂O, and 1.0 g/l Tween 80. The OM significantly improved the biomass production of JW15 over an established commercial medium (MRS). After fermenting OM, the dry cell weight of JW15 was 4.89 g/l, which was comparable to the predicted value (4.77 g/l), and 1.67 times higher than that of the MRS medium (3.02 g/l). Correspondingly, JW15 showed a rapid and increased production of lactic and acetic acid in the OM. To perform a scale-up validation, batch fermentation was executed in a 5-l bioreactor at 37℃ with or without a pH control at 6.0 ± 0.1. The biomass production of JW15 significantly improved (1.98 times higher) under the pH control, and the cost of OM was reduced by two-thirds compared to that in the MRS medium. In conclusion, OM may be utilized for mass producing JW15 for industrial use.

• Applied Biological Chemistry
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• v.10
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• pp.69-100
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• 1968

1. In order to investigate on the microflora and enzyme activity of mold wheat 'Nuruk' , the major source of microorganisms for the brewing of Takju (a Korean Sake), two samples of Nuruk, one prepared at the College of Agriculture, Chung Nam University (S) and the other perchased at a market (T), were taken for the study. The molds, aerobic bacteria, lactic acid bacteria, and yeasts were examined and counted. The yeasts were classified by the treatment with TTC (2, 3, 5 triphenyltetrazolium chloride) agar that yields a varied shade of color. The amylase and protease activities of Nuruk were measured. The results were as the followings. a) In the Nuruk S found were: Aspergillus oryzae group, $204{\times}10^5$; Black Aspergilli, $163{\times}10^5$; Rhizogus, $20{\times}10^5$; Penicillia, $134{\times}10^5$; Areobic bacteria, $9{\times}10^6-2{\times}10^7$; Lactic acid bacteria, $3{\times}10^4$ In the Nuruk T found were: Aspergillus oryzae group, $836{\times}10^5$; Black Aspergilli, $286{\times}10^5$; Rhizopus, $623{\times}10^5$; Penicillia, $264{\times}10^5$; Aerobic bacteria, $5{\times}10^6-9{\times}10^6$; Lactic acid bacteria, $3{\times}10^4$ b) Eighty to ninety percent of the aerobic bacteria in Nuruk S appeared to belong to Bacillus subtilis while about 70% of those in Nuruk T seemed to be spherical bacteria. In both Nuruks about 80% of lactic acid bacteria were observed as spherical ones. c) The population of yeasts in 1g. of Nuruk S was about $6{\times}10^5$, 56.5% of which were TTC pink yeasts, 16% of which were TTC red pink yeasts, 8% of which were TTC red yeasts, 19.5% of which were TTC white yeasts. In Nuruk T(1g) the number of yeasts accounted for $14{\times}10^4$ and constituted of 42% TTC pink. 21% TTC red pink 28% TTC red and 9% TTC white. d) The enzyme activity of 1g Nuruk S was: Liquefying type Amylase, $D^{40}/_{30},=256$ W.V. Saccharifying type Amylase, 43.32 A.U. Acid protease, 181 C.F.U. Alkaline protease, 240C.F.U. The enzyme activity of 1g Nuruk T was: Liquefying type Amylase $D^{40}/_{30},=32$ W.V. Saccharifying type amylase $^{30}34.92$ A.U. Acid protease, 138 C.F.U. Alkaline protease 31 C.F.U. 2. During the fermentation of 'Takju' employing the Nuruks S and T the microflora and enzyme activity throughout the brewing were observed in 12 hour intervals. TTC pink and red yeasts considered to be the major yeasts were isolated and cultured. The strains ($1{\times}10^6/ml$) were added to the mashes S and T in which pH was adjusted to 4.2 and the change of microflora was examined during the fermentation. The results were: a) The molds disappeared from each sample plot since 2 to 3 days after mashing while the population of aerobic bacteria was found to be $10{\times}10^7-35{\times}10^7/ml$ inS plots and $8.2{\times}10^7-12{\times}10^7$ in plots. Among them the coccus propagated substantially until some 30 hours elasped in the S and T plots treated with lactic acid but decreased abruptly thereafter. In the plots of SP. SR. TP. and TR the coccus had not appeared from the beginning while the bacillus showed up and down changes in number and diminished by 1/5-1/10 the original at the end stage. b) The lactic acid bacteria observed in the S plot were about $7.4{\times}10^7$ in number per ml of the mash in 24 hours and increased up to around $2{\times}10^8$ until 3-4 days since. After this period the population decreased rapidly and reached about $4{\times}10^5$ at the end, In the plot T the lactic acid becteria found were about $3{\times}10^8$ at the period of 24 fours, about $3{\times}10$ in 3 days and about $2{\times}10^5$ at the end in number. In the plots SP. SR. TP, and TR the lactic acid bacteria observed were as less as $4{\times}10^5$ at the stage of 24 hours and after this period the organisms either remained unchanged in population or ceased to exist. c) The maiority of lactic acid bacteria found in each mash were spherical and the change in number displayed a tendency in accordance with the amount of lactic acid and alcohol produced in the mash. d) The yeasts had showed a marked propagation since the period of 24 hours when the number was about $2{\times}10^8$ ㎖ mash in the plot S. $4{\times}10^8$ in 48 hours and $5-7{\times}10^8$ in the end period were observed. In the plot T the number was $4{\times}10^8$ in 24 hours and thereafter changed up and down maintaining $2-5{\times}10^8$ in the range. e) Over 90% of the yeasts found in the mashes of S and T plots were TTC pink type while both TTC red pink and TTC red types held range of $2{\times}10-3{\times}10^7$ throughout the entire fermentation. f) The population of TTC pink yeasts in the plot SP was as $5{\times}10^8$ much as that is, twice of that of S plot at the period of 24 hours. The predominance in number continued until the middle and later stages but the order of number became about the same at the end. g) Total number of the yeasts observed in the plot SR showed little difference from that of the plot SP. The TTC red yeasts added appeared considerably in the early stage but days after the change in number was about the same as that of the plot S. In the plot TR the population of TTC red yeasts was predominant over the T plot in the early stage which there was no difference between two plots there after. For this reason even in the plot w hers TTC red yeasts were added TTC pink yeasts were predominant. TTC red yeasts observed in the present experiment showed continuing growth until the later stage but the rate was low. h) In the plot TP TTC pink yeasts were found to be about $5{\times}10^8$ in number at the period of 2 days and inclined to decrease thereafter. Compared with the plot T the number of TTC pink yeasts in the plot TP was predominant until the middle stage but became at the later stage. i) The productivity of alcohol in the mash was measured. The plot where TTC pink yeasts were added showed somewhat better yield in the earely stage but at and after the middle stage the difference between the yeast-added and the intact mashes was not recognizable. And the production of alcohol was not proportional to the total number of yeasts present. j) Activity of the liquefying amylase was the highest until 12 hours after mashing, somewhat lowered once after that, and again increased around 36-48 hours after mashing. Then the activity had decreased continuously. Activity of saccharifying amylase also decreased at the period of 24 hours and then increased until 48 hours when it reached the maximum. Since, the activity had gradually decreased until 72 hours and rapidly so did thereafter. k) Activity of alkaline protease during the fermentation of mash showed a tendency to decrease continusously although somewhat irregular. Activity of acid protease increased until hours at the maximum, then decreased rapidly, and again increased, the vigor of acid protease showed better shape than that of alkaline protease throughout. 3. TTC pink yeasts that were predominant in number, two strains of TTC red pink yeasts that appeared throughout the brewing, and TTC red yeasts were identified and the physiological characters examined. The results were as described below. a) TTC pinkyeasts (B-50P) and two strains of TTC red pink yeasts (B-54 RP & B-60 RP) w ere identified as the type of Saccharomyces cerevisiae and TTC pink red yeasts CB-53 R) were as the type of Hansenula subpelliculosa. b) The fermentability of four strains above mentioned were measured as follows. Two strains of TTC red pink yeasts were the highest, TTC pink yeasts were the lowest in the fermantability. The former three strains were active in the early stage of fermentation and found to be suitable for manufacturing 'Takju' TTC red yeasts were found to play an important role in Takju brewing due to its strong ability to produce esters although its fermentability was low. c) The tolerance against nitrous acid of strains of yeast was marked. That against lactic acid was only 3% in Koji extract, and TTC red yeasts showed somewhat stronger resistance. The tolerance against alcohol of TTC pink and red pink yeasts in the Hayduck solution was 7% while that in the malt extract was 13%. However, that of TTC red yeasts was much weaker than others. Liguefying activity of gelatin by those four strains of yeast was not recognized even in 40 days. 4. Fermentability during Takju brewing was shown in the first two days as much as 70-80% of total fermentation and around 90% of fermentation proceeded in 3-4 days. The main fermentation appeared to be completed during :his period. Productivity of alcohol during Takju brewing was found to be apporximately 65% of the total amount of starch put in mashing. 5. The reason that Saccharomyces coreanuss found be Saito in the mash of Takju was not detected in the present experiment is considered due to the facts that Aspergillus oryzae has been inoculated in the mold wheat (Nuruk) since around 1930 and also that Koji has been used in Takju brewing, consequently causing they complete change in microflora in the Takju brewing. This consideration will be supported by the fact that the original flavor and taste have now been remarkably changed.