• Applied Biological Chemistry
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• v.11
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• pp.35-41
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• 1969

In order to determine proper ratio of wheat to soybean of soysauce koji material for Korean people's taste, sensory evaluation for the soysauces made from soysauce kojis with various above-mentioned ratio and analysis of chemical components through the brewing period of six months were carried out. The results obtained were summarized as follows: 1. The specific gravity of all the soysauces increased gradually as the time passed and had maximum values in two months with nearly constant values thereafter. Concerning soybean/wheat-ratio, generally, the higher soybean/wheat-ratio, the lower specific gravity was observed but when this ratio was higher than 10 : 6, the gravity was on the similar levels. 2. The solid material in all the soysauces increased in the earlier stage and there were no significant differences among the treatments but soysauces without wheat had a little low content. 3. The lesser amount of wheat as raw material was, the higher contents of total nitrogen were and the contents of total nitrogen in the every ratio of raw material increased parallely as the mash ages. 4. The amino-nitrogen levels were found to have the simllar tendency to total nitroghen levels. 5. The reducing sugar content was higher in the soysauces with more-wheat and increased rapidly up to the maximum level in two months and then the higher wheat/soybean-ratio was, the slower decrease of the sugar content was observed. 6. The maximum total acidity was observed in the soysauces with 10; 4-6 (soybean/wheat-ratio) and increased similary in all the soysauces throughout the period. 7. According to the sensory evaluation, the short (2-3 months) fermented soysauces showed best taste when soybean/wheat-ratio was 10 : 8 and 10 : 6 if diluted, but in long (4-6 months) fermented soysauces it was 10 : 6 and 10 : 4 if diluted.

• Food Science and Preservation
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• v.18 no.3
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• pp.397-406
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• 2011

To industrialize meju, four kinds of meju (Korean-style soybean koji) were made with humidity-controlled fermentation for 12 days at $28^{\circ}C$ after they were inoculated with selected strains such as Aspergillus otyzae (AO meju), Aspergillus sojae (AS meju), combined Aspergillus sojae and Bacillus subtilis (ASBS meju), and combined Aspergillus oryzae and Bacillus subtilis (AOBS meju) as starter microorganisms. The changes in the quality characteristics in the four kinds of meju were investigated during fermentation. Their enzyme activities were compared with those of the traditional meju that is made in Sunchang Folk Village according to the traditional method. In the meju that were inoculated with selective strains, the aerobic bacteria counts and mold counts exceeded 8 log cfu/g and 6 log cfu/g respectively, which were the highest fermentation values after 2 days. The aerobic bacteria counts were maintained from 2-day to the 12-day fermentation. The mold counts tended to decreased gradually after the 2-day fermentation. The amino-type nitrogen contents reached 430.5-577.5 mg%, which were the highest values after 2-day fermentation. The neutral protease activities of these mejus had the highest levels in the following order: traditional meju, $1,258.0{\pm}38.8$; AS meju, $1,238.3{\pm}38.6$; AO meju. $1,204.1{\pm}24.1$; ASBS meju, $1,040.6{\pm}10.6$; and AOBS meju, $1,033.5{\pm}11.2$ unit/g. The acidic protease activities of these meju had the highest levels in the following order: AO meju, $1,030.1{\pm}19.1$, traditional meju, $1,007.7{\pm}30.5$; AS meju, $990.9{\pm}25.0$; AOBS meju, $910.9{\pm}15.3$; and ASBS meju, $888.2{\pm}15.7$ unit/g.

• Food Science and Preservation
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• v.17 no.5
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• pp.741-751
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• 2010

We evaluated the nutritional value of a Monascus pilosus mycelial ethanolic extract (MEM) and culture filtrate (CFM) by determining the contents of monacolin K and citrinin, and by measuring antioxidant and antimicrobial activities. The yields of freeze-dried MEM and CFM powder were 4.02% and 3.35% of wet weight, respectively. Pigment content ($OD_{500}$ value) of MEM (0.79) and CFM (0.63) were lower than those of commercial rice beni-koji ethanolic extracts (EERB) (0.87), but were in good agreement with the L*, a*, and b* values and the hue angles of the products. The total monacolin K content of MEM (24.91 mg%) was higher than those of CFM (1.27 mg%) and EERB (14.65 mg%). However, the active monacolin K content of EERB (5.48 mg%) was higher than those of MEM (3.35 mg%) and CFM (0.4 mg%). Citrinin was not detected in any sample. The total polyphenol content of MEM (4.68%, w/w) was similar to that of CFM (4.29%, w/w), thus 13.75.20.94% higher than that of EERB. The total flavonoid content of EERB was 6.8.7.0-fold higher than those of MEM (0.64%, w/w) and CFM (0.66%, w/w). The total antioxidant capacity of CFM (3.51%, w/w) was 1.62.2.08-fold higher than those of MEM (2.74%, w/w) and EERB (1.69%, w/w). The electron-donating capacities of 1% (w/v) solutions of CFM, MEM, BHT, and EERB were 86.20%, 77.25%, 77.25%, and 44.82%, respectively, and the corresponding reducing powers ($OD_{700}$ values) were 2.1, 2.4, 1.1, and 1.6, respectively. SOD(superoxide dismutase)-like activities were in the order MEM (39.85%) > BHT (37.68%) > EERB (26.70%) > CFM (21.5%). Although the TBARS (% value) of MEM was a little lower than that of BHT, it was higher than those of CFM and EERB. The antibacterial activities of CFM acting on Bacillus brevis and Escherichia coli were somewhat higher than those of MEM, whereas the activities of MEM on Bacillus subtilis, Listeria monocytogenes, Staphylococcus aureus, Staphylococcus epidermidis, and Salmonella enteritidis were higher than those of CFM. However, the antibacterial activities of MEM and CFM were less than those of EERB and BHT. In conclusion, although further studies are needed, we offer experimental evidence that the by-products of M. pilosus MEM and CFM contain significant antioxidant and antimicrobial activities that may be useful in the development of healthy foods.

• Applied Biological Chemistry
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• v.17 no.2
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• pp.81-92
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• 1974

In order to prepare the mashing materials for 'Takju', Korean wine, with potatoes, theywere steamed, dryed, and pulverized, and their chemical components were analyzed. As a brewing method of Takju with potatoes, general 2nd stage process with Ipkuk and Bunkuk (enzyme sources), commonly used now, was carried out and the effects of preparing conditions of Ipkuk(koji) with potato flour, mashing materials and brewing conditions on the contents of Takju mash, and of storing time on the contents of Takju, were investigated and the results obtained were summarized as follows, 1. Chemical components of steamed potatoes and potato flour were Moisture; 76.2, 10.8%, Total sugar; 16.1, 69.8%, Reducing sugar; 3.45, 13.4%, Crude protein; 2.1, 11.3%, Total acid; 0.012, 0.023% and Volatile acid; 0.0012, 0.0025% respectively. 2. The most effective preparing conditions of Ipkuk with potato flour were to incubate the potato flour added 40-50% of water for 48 hours by general preparing process of Koji, and liquefying and saccharogenic amylase activities of Ipkuk incubated at above conditions were $D40^{\circ}$ 30' 128 W.V. and 13.2 A.U.. 3. The effects of various brewing conditions on the contents of Takju mashes were as follows; 1) Optimum ratio of mashing water and materials for Takju brewing with potato flour was 140ml of water to 60g of flour in 1st stage and 260ml to 140g in 2nd stage. 2) Optimum fermentating times and temperatures for Takju brewing were at $25^{\circ}C$ for 48 hours in 1st stage and at $30^{\circ}C$ for 48 hours in 2nd stage. 3) Optimum amounts of enzyme sources for Takju brewing were 20-30% of Ipkuk and 0.5% of Bunkuk in 1st stage and 1.0% of Bunkuk in 2nd stage. 4) Methanol content of the Takju mash brewed with raw potato flour was much more than that with steamed potato flour. 5) Alcohol, fusel oil and Formol nitrogen contents of the Takju mash brewed with potato flour were less than that with wheat flour, on the contrary, methanol contents and total acidifies of them were showed conversely above. 4. The changes of chemical components and microflora in the mashes during the brewing potato flour Takju were as follows; 1) The accumulation of ethanol followed rapidly in early stage, being the highest at 72 hours (11.9%). 2) Total sugar content of the mash was decreased considerably within 48-72 hours, being 2.62% at 72 hours, and thereafter slowly. 3) Reducing sugar of the mash had a tendency of decreasing, being 0.29% at 48 hours. 4) Total acidity, volatile acidity and Formol nitrogen content of the mash were increased slowly, being 7.30, 0.20, 2.55 at 48 hours. 5) Total cells of yeast appeared the highest in 72 hours ($2.1{\times}10^8$) and thereafter decreased slowly. 6) Total cells of bacteria appeared the highest in 48 hours ($2.4{\times}10^8$) and thereafter decreased or increased slightly. 5. Takju was made from the fermented mash mixed with water to be 6% of alcohol content, and the change of alcohol content, total acidity, total cells of yeast and bacteria during the storing at $30^{\circ}C$ were as follows; 1) Alcohol content of Takju was increased slightly at 24 hours (6.2%), and thereafter decreased slowly. 2) Total acidity of Takju was increased gradually, being 6.1 at 72 hours 3) Total cells of yeast and bacteria appeared the highest at 48 hours ($2.3{\times}10^8,\;1.5{\times}10^8$), and thereafter decreased slowly. 6. Alcohol content, total acidity and Formol nitrogen content of the Takju brewed with potato flour Ipkuk or wheat flour Ipkuk and steamed potatoes(1:5) were 9.8-11.3%, 5.8-7.4, 2.5-3.3 respectively, and the color of the Takju. was similar to commercial Takju. 7. The results of sensory test for various experimental Takju, showed that the Takjues brewed with the materials combined with wheat flour and steamed potatoes(4:5 or 3.5:7.5) were not significantly different in color, taste and flavor from commercial Takju, However, those with potato flour and wheat flour (1:1 or 7:3) were significantly different from commercial Takju.

• Korean Journal of Agricultural Science
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• v.1 no.1
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• pp.67-81
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• 1974

In order to prepare the mashing materials for "Takju", Korean wine, with potatoes they were steamed, dryed, and pulverized, and their chemical components were analyzed. As a brewing method of Takju with potatoes, general 2nd stage process with Ipkuk and Bunkuk (enzyme sources), commonly used now, was carried out and the effects of preparing conditions of Ipkuk(koji) with potato flour, mashing materials and brewing conditions on the contents of Takju mash and of storing time on the contents of Takju, were investigated and the results obtained were summarized as follows. 1. Chemical components of steamed potatoes and potato flour were Moisture; 76.2, 10.8%, Total sugar; 16.1, 69.8%, Reducing sugar; 3.45, 13.4%, Crude protein; 2.1, 11.3%, Total acid; 0.012, 0.023% and Volatile acid; 0.0012, 0.0025% respectively 2. The most effective preparing conditions of Ipkuk with potato flour were to incubate the potato flour added 40-50% of water for 48 hours by general preparing process of Koji, and liquefying and saccharogenic amylase activities of Ipkuk incubated at above conditions were $D_{40^{\circ}}{^{30{\prime}}}$ 128 W.V. and 13.2 A. U. 3. The effects of various brewing conditions on the contents of Takju mashes wereas follows; 1) Optimum ratio of mashing water and materials for Takju brewing with potato flour was 140ml of water to 60g of flour in 1st stage and 260ml to 140g in 2nd stage. 2) Optimum fermentating times and temperatures for Takju brewing were at $25^{\circ}C$ for 48 hours in 1st stage and at $30^{\circ}C$ for 48 hours in 2nd stage. 3) Optimum amounts of enzyme sources for Takju brewing 20-30% of Ipkuk and 0.5% of Bunkuk in 1st stage and 1.0% of Bunkuk in 2nd stage. 4) Methanol content of the Takju mash brewed with raw potato flour was much more than that with steamed potato flour. 5) Alcohol fusel oil and Formal nitrogen contents of the Takju mash brewed with potato flour were less than that with wheat flour, on the contrary, methanol contents and total acidities of them were showed conversely above. 4. The changes of chemical components and microflora in the mashes during the brewing potato flour Takju were as follows; 1) The accumulation of ethanol followed rapidly in early stage, being the highest at 72 hours (11.9%) 2) Total sugar content of the mash was decreased considerably within 48-72 hours, being 2.62% at 72 hours, and thereafter slowly. 3) Reducing sugar of the mash had a tendency of decreasing, being 0.29% at 48 hours. 4) Total acidity, volatile acidity and Formal nitrogen content of the mash were increased slowly, being 7.30, 0.20, 2.55 at 48 hours. 5) Total cells of yeast appeared the highest in 72 hours ($2.1{\times}10^8$) and thereafter decreased slowly. 6) Total cells of bacteria appeared the highest in 48 hours ($2.4{\times}10^8$) and thereafter decreased or increased slightly. 5. Takju was made from the fermented mash mixed with water to be 6% of alcohol content, and the change of alcohol content, total acidity, total cells of yeast and bateria during the storing at $30^{\circ}C$ were as follows; 1) Alcohol content of Takju was increased slightly at 24 hours (6.2%), and thereafter decreased slowly. 2) Total acidity of Takju was increased gradually, being 6.1 at 72 hours 3) Total cells of yeast and bacteria appeared the highest at 48 hours ($2.3{\times}10^8$, $1.5{\times}10^8$) and thereafter decreased slowly. 6. Alcohol content, total acidity and Formol nitrogen content of the Takju brewed with potato flour Ipkuk or wheat flour Ipkuk and steamed potatoes(1:5) were 9.8-11.3%, 5.8-7.4, 2.5-3.3 respectively, and the color of the Takju was similar to commercial Takju. 7. The results of sensory test for various experimental Takju, showed that the Takjues brewed with the materials combined with wheat flour and steamed potatoes (4:5 or 3.5:7.5) were not significantly different in color, taste and flavor from commercial Takju, However, those with potato flour and wheat flour (1:1 or 7:3) were significantly different from commercial Takju.

• 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.