• Applied Biological Chemistry
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• v.18 no.1
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• pp.23-29
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• 1975

This experiment was conducted to study the effects of temperature and pH upon the acid productivity of the acid producing mutant induced by the treatment of ultraviolet light, and to identify the producing acid by PPC and p-oxydiphenyl method. Chemical composition of Takju mash brewed with selected yeast and producing acid were observed and the results were as follows. 1) There was no apprecible difference in acid producing activity of mutant at $25^{\circ}C\;to\;30^{\circ}C$. 2) The acid producing activity of mutant was little below pH 4 and was gradually increased according to approach nenutral, and the accumulation of acid was amounted to 0.5-0.7% as a lactic acid at pH 5 to 7 within 48 hrs of fermentation. 3) The acid produced by mutant was detected to the lactic acid. 4) In the cases of the Takju was brewed with the starter from the acid producing mutant the requirement of Ipkuk was 5% for all the raw materials, on the contrary, using orginal strain the requirement of Ipkuk was 20%. 5) In the case of both starters from the acid producing mutant and orginal strain were added at different brewing times, and only Bunkuk was used as a saccharifying agent (without Ipkuk), Takju was able to brewed more repidly and successfully than the case of general process.

• Korean Journal of Food Science and Technology
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• v.34 no.2
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• pp.296-302
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• 2002

Quality characteristics of mash of takju prepared by wheat flour nuruks (conventional and improved style) were investigated during fermentation. Those were cultured with wild microorganism on whole and crushed wheat (wheat nuruk), wheat flour and mixture of rice and wheat flour, and nuruk cultured with Aspergillus oryzae on wheat flour. Ethanol content reached maximum of 10.6-17.4% after 14 days of fermentation. And the ethanol contents of takju were high in order of wheat nuruk, wheat flour nuruk, Aspergillus oryzae nuruk and mixture of wheat flour and rice nuruk. No difference of pH was among the treatments while the wheat flour nuruk cultured with wild microorganisms contained higher amounts of acids than the others. The amount of total sugars ranged from 16.22-17.74% on the starting day of fermentation, and decreased to 4.28-6.10% after 14 days. The amount of total sugars in mashes brewed with wheat flour nuruk cultured with Aspergillus oryzae was measured to be highest at the beginning stage of fermentation. Afterwards no difference was found among the types of nuruks. Glucose was in the range of 2,735-7,842 mg% at initial period of fermentation, afterwards it was rapidly decreased. The total amounts of free sugar for control and the wheat flour nuruk was higher than other treatments. The total amounts of organic acids were the highest in wheat nuruk to 690-2,241 mg% and the contents of lactic acid and succinic acid were high in mixed rice nuruk to 183-1,293 mg%. The contents of lactic acid and succinic acid were higher than those of the other kinds of organic acids in takju.

• Applied Biological Chemistry
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• v.13 no.3
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• pp.219-222
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• 1970

Riboflavin in the various materials which used for Takju brewing was determined and its quantitative changes in the Takju mashes which were mashed with polished rice and wheat flour respectively using Nuruk only for enzyme source, and with wheat flour and corn powder respectively using Nuruk and Koji, were checked at 24 hours intervals during the whole brewing period. The results obtained were as follows. a) The riboflavin contents in 100 grams of each materials were: Polished rice $46.9{\gamma}$ Wheat flour $67.3{\gamma}$ Corn powder $73.9{\gamma}$ Nuruk $169.0{\gamma}$ Koji of wheat flour $87.1{\gamma}$ Koji of corn powder $84.4{\gamma}$ b) About 30 to 40 percent of riboflavin in the polished rice, wheat flour and corn powder were losed by steaming at 10 pounds for 50 minutes. c) Riboflavin content in the plots used Nuruk only was decreased during 3 days after mashing and then increased markedly after 5 days and, more or less, decreased after 6 days. Quantitative changes of riboflavin in the plots used Nuruk and Koji showed a rugged curve without decreasing in early stages, and its content was also increased markedly 5 days after mashing and then decreased a little after 6 days. d) Approximately $82{\gamma}$ of riboflavin was determined in 100ml. of the ripened mash of the rice plot and $72{\gamma}$ in the wheat flour plot in the case of using Nuruk only. $88.6{\gamma}$ of riboflavin were determined in 100ml. of the ripened mash of the wheat flour plot and $83.6{\gamma}$ in the corn powder plot in the case of using Nuruk and Koji. e) It was assumed that $28.8{\gamma}$ to $41{\gamma}$ of riboflavin were contained in 100 ml. of finished Takju of the two plots used Nuruk only and 33.6 to 44.5${\gamma}$ in the both two plots used Nuruk and Koji. f) About $47.3{\gamma}$ of riboflavin were determined in 100 grams of Takju cake of the rice plot and $66.2{\gamma}$ in the wheat flour plot used Nuruk only, and $62.9{\gamma}$ in the corn powder plot and $64.5{\gamma}$ in the wheat flour plot used Nuruk and Koji.

• Applied Biological Chemistry
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• v.13 no.1
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• pp.105-109
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• 1970

Thiamin in the various materials which used for Takja braving was determined and its quantitative changes in the Takju mashes which were mashed with polished rice and wheat flour respectively using Nuruk only for the enzyme source, and with wheat flour and corn powder respectively using Nuruk and Koji, were checked at 24 hours intervals during the whole brewing period. The results obtained were as follows. a) The thiamin contents in 100 grams of each materials were: Polished rice 107.8${\gamma}$ Wheat Pour 185.0${\gamma}$ Corn powder 410.2${\gamma}$ Nuruk 347.4${\gamma}$ Koji of wheat flour 170.1${\gamma}$ Koji of corn powder 257.3${\gamma}$ b) About 40 to 50 percent of thiamin in the polished rice, wheat flour and, corn powder were losed by steaming at 10 wounds for 50 minutes. c) The quantitative changes of thiamin in the Takju meshes showed a similar tendency. Thiamin contents were decreased markedly during 2 days after mashing but thereafter, almost unchanged till the mashes were ripened. Its decreasing rates in tile two plots uses Nuruk and Koji wereless thaw those in the two plots used Nuruk only. d) Approximately 6${\gamma}$ of thiamin was determined in 100 ml. of the ripened mash of the rice plot and 7.5${\gamma}$ in tire whet flour plot in the case of using Nuruk only. 12.4${\gamma}$ of thiamin was determined in 100 ml. of the ripened mash of the wheat flour plot and 15.4${\gamma}$in the corn powder plot in the case of using Nuruk and Koji. e) It was assumed that 2.4 to 3.5${\gamma}$ of thiamin were contained in 100 ml. of finished Takju of the two plots used Nuruk only and 5 to 7${\gamma}$ in the two plots used Nuruk and Koji. f) About 43.7${\gamma}$ of thiamin were determined in 100 grams of Takju cake of the rice plot and 56.1${\gamma}$ in the wheat flour plat used Nuruk only, and 81.4${\gamma}$ in the corn powder plot and 57.7${\gamma}$ in the wheat flour plot used Nuruk and Koji.

• Korean Journal of Agricultural Science
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• v.4 no.2
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• pp.173-177
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• 1977

These experiments were conducted to investigate the formation of organic acid and ethanol during fermentation by three yeast strains. One of these was industrial strain (No.7) from Japan, and the others were wild types (No. 47, No. 239) isolated from Takju-mash and Strawbery, respectively. Conditions of fermentation were varied by differing the supply of oxygen (air), and by using different fermentation media The results obtained were as follows: 1) All the yeast strains produced higher amount of total organic acid and ethanol under the conditions which were aerobic, i.e. the flasks were opened during fermentation, than in case of using the flasks with fermentation bung. 2) Organic acid and ethanol were produced rapidly in the mash medium than in the semi-synthetic medium, i.e. total amount of organic acid and ethanol was maximized in a short time in the mash medium. 3) On the mash medium, the highest amount of organic acid was obtained by the strain No. 239, the next by No. 7 and the lowest by No. 47. Ethanol was produced on this medium with decreacing order of No. 47, No. 239, and No. 7. 4) The strain No. 239 was proved to be a powerful organic acid producer, yielding higher amount of organic acid especially under the aerobic conditions. 5) Above results suggests that the strain No. 239 could be of useful in alcoholic drink industry, due to its powerful ethanol-producing characteristic accompaning with high yielding of organic acids.

• 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.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.744-750
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• 1999

Forty alcohol yeast strains were isolated from the main mashes (10 strains from each mash) for brewing of 4 different kinds of Korean traditional liquor (3 different types of Yakju and 1 Takju). Thirty-eight out of 40 strains were identified to be the same strain, Saccharomyces boulardii, by the Automated Bacteria, Yeast, and Fungi Identification System (Biolog Co., U.S.A.) based on the metabolic fingerprints. One strain that showed the highest ethanol production among the 38 strains in YPD medium, designated SHY 111, was selected and used for differentiating from other yeast type strains using the polymerase chain reaction (PCR). Amplified DNA, from transcribed internal spacers of SHY 111 chromosomal DNA, was found to be the same in both size and sequence as those of S. cerevisiae KCCM 11215 (formerly S. coreanus) and S. boulardii along with that of S. cerevisiae AB 972, which was used as a type strain for the yeast genome project. However, when PCR was carried out with the intron splice site primer, it resulted in the amplification of the SHY 111-specific DNA fragment which was about 200 bp in size. When PCR was carried out using the primer to test diversity of 40 isolated yeast strains, it was found that the PCR patterns were similar to each other except for the 200 bp bands derived from all the 10 strains from one Yakju, and 2 strains from another Yakju. These results suggest the strain identified as S. boulardii by the Automated Identification System to be a dominant strain for the fermentation of Korean traditional liquors.

• Korean Journal of Food Science and Technology
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• v.42 no.1
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• pp.56-62
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• 2010

Quality characteristics such as alcohol, acid, and sugar contents of takju brewing mashes prepared using several yeasts were investigated during 12-days of fermentation. Among the yeasts examined, S. cerevisiae led to the highest level of ethanol (10.2-13.4%) and total minor alcohols (0.729-0.831 mg/mL). Regardless of the yeasts used, the acidity showed drastic changes (pH from 3.4-4.2 down to 2.4-2.7) during the first 2 days, and displayed negligible changes from day 4. The total acid contents rapidly increased to the first 4 days of fermentation and leveled off from 4 to the end of fermentation period. The common organic acid components were lactic, succinic and acetic acids. The total and reducing sugar contents varied depending on the yeasts used, with mashes prepared using S. coreanus and S. rouxii producing the maximum total sugar contents (5.43-5.5%) at the end of fermentation. The reducing sugar showed its maximum (7.53-14.89%) at day 2, after which it decreased to its minimum levels (3.04-4.52%). The common free sugar components were glucose and fructose, while S. ellipsoideus led to a higher free sugar level (0.35-5.29%).

• 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.