• 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 Food Hygiene and Safety
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• v.21 no.2
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• pp.100-106
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• 2006

The use of underarm and body care cosmetics with oestrogenic chemical excipients (particularly the parabens) and the hypothesized association with breast cancer incidence, particularly in women. It is noted that the type of cosmetic product is irrelevant (e.g. antiperspirant/deodorant versus body lotion, moisturizers or sprays versus creams) and attention must focus on issues of actual exposure to chemicals through continued dermal application of body care products and the endocrine/hormonal activity and toxicity of the chemicals in the formulations. To evaluate the estrogenic activities of parabens such as ethylparaben, butylparaben, propylparaben, isobutylparaben and isopropylparaben, we used recombinant yeasts containing the human estrogen receptor [Saccharomyces cerevisiae ER+LYS 8127], human breast cancer MCF-7 cell lines and human estrogen receptor ${\alpha}\;and\;{\beta}$. In E-screen assays, isopropylparaben is the most estrogenic paraben, and in ER competition assay, isobutylparaben is the most estrogenic paraben. We evaluated isopropylparaben was most active in the recombinant yeast assay, followed by propylparaben, ethylparaben, isobutylparaben and butylparaben. Results from this study demonstrate that parabens are observed in human endocrine system. Therefore, we have shown that the parabens is induced the estrogenic activities similar to $17{\beta}$-estradiol and Bisphenol-A.

• Journal of the Korean Society of Food Culture
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• v.18 no.6
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• pp.551-561
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• 2003

To determine the conditions of the fermentation and storage for Chonggak kimchi in kimchi refrigerator, prepared Chonggak kimchi took into kimchi refrigerators which were controlled at four different modes of the fermented temperature and time, and fermented and kept for 16 weeks. The pH in Chonggak kimchi fermented at $20^{\circ}C$ for 24 hours/stored at $-1^{\circ}C$ dropped greater than all of kimchi fermented at other combinations, and the changes of pH at any combinations were not greater than those in Baechu kimchi, because pH in Chonggak kimchi did not dropped below 4.5. Acidities in Chonggak kimchi were greatly increased at higher temperature. The acidity in Chonggak kimchi during the first week of fermentation was lower than that in Baechu kimchi and then it was rather higher because of the addition of waxy rice paste. In texture, puncture force of Chonggak kimchi was decreased slowly until 8 weeks of fermentation and then did not changed much and the highest values showed in Chonggak kimchi stored directly at $-1^{\circ}C$ without any fermentation. In sensory evaluation, the scores for the carbonated flavor and the sourness were the highest in Chonggak kimchi fermented at $20^{\circ}C$ for 24 hours/stored at $-1^{\circ}C$, but the lowest in Chonggak kimchi stored directly at $-1^{\circ}C$ without any fermentation because of some undesirable flavors. The lowest hardness showed in Chonggak kimchi fermented at highest temperature and the best hardness was in Chonggak kimchi fermented at $5^{\circ}C$ for 3 days or 6 days/stored at $-1^{\circ}C$. The appearance was the best in Chonggak kimchi fermented at $20^{\circ}C$ for 24 hours/stored at $-1^{\circ}C$ and the worst was in Chonggak kimchi stored directly at $-1^{\circ}C$ without any fermentation. The overall acceptability of Chonggak kimchi fermented at $20^{\circ}C$ for 24 hours/stored at $-1^{\circ}C$ was good after 4 weeks of fermentation, but in Chonggak kimchi fermented at $5^{\circ}C$ for 3 days or 6 days/stored at $-1^{\circ}C$ it was good after 6 weeks. Total microbial counts in most of Chonggak kimchi were reached to a maximum number within 7 days, and then decreased similarly at all modes. Leuconostoc spp. and Lactobacillus spp. increased to maximum number of $1.48{\times}10^9\;and\;5.62{\times}10^9$, respectively, in Chonggak kimchi fermented for 7 days. Yeast counts showed a increasing trend not depends on fermenting temperature and they were lower counts than those in Baechu kimchi. Waxy rice paste which added to Chonggak kimchi resulted in increasement of glucose as a carbon source and stimulated to reproduce the microbes in Chonggak kimchi.

• Applied Biological Chemistry
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• v.18 no.1
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• pp.16-22
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• 1975

This experiment was carried out to investigate the physiological characteristics of two original yeasts, 5-Y-5 and 6-Y-6, which selected from 24 Takju yeasts and three mutants, 30-24,30-81 and 40-27. induced from two original yeasts by the irradiation of UV light. The results were summarized as follows. 1) Alcohol tolerances of three mutants were decreased in some degree as compared with those of original yeasts. 2) Tolerances of lactic and citric acids of acid producing mutant 30-81, was increased than those of original yeasts. 3) In the case of using ammonium sulfate as a nitrogen source, two original yeasts and three mutants required Ca-pantothenate as a essential growth factor and four strains of yeasts except the mutant, 30-81, required biotin as a stimulated growth factor, When asparagine was used as a nitrogen source, two original yeasts and three mutants showed the same as above result but the stimulated effect of biotin was far less. 4) Propagation powers of the mutants were weaken than those of original yeasts, particular that of acid producing mutant, 30-81, was the weakest in the three mutants. 5) The optimum temperature for fermentation of original yeasts were $30^{\circ}C\;to\;35^{\circ}C$ but three mutants were $25^{\circ}C\;to\;30^{\circ}C$. 6) The optimum pH for fermentation of original yeasts were pH 5 to 6, and there is no appreciable difference between original yeasts and three mutants. The fermentation power of mutant,30-81, was decreased more rapidly than those of other mutants according to approach neutral. Three mutants were more sensible to heat than original yeasts. 7) Two original yeasts and three mutants were inhibited more over 20 percent of sugar for fermentation and three mutants were more sensible to sugar concentration than original yeasts.

• Journal of The Korean Society of Grassland and Forage Science
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• v.37 no.4
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• pp.290-300
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• 2017

In vitro fermentation was conducted to figure out alternative fiber sources for horse feed. For the development of value-added products as a horse feed resource, the pomaces from apple, carrot, grape, and citrus were fermented under solid-state conditions in the presence of 60% soybean meal with 40% of each fruit pomace at 60% of moisture content. Lactobacillus plantarum SK3873, Lactobacillus plantarum SK3893, Weissella cibaria SK3880, and Bacillus subtilis SK3889 were isolated from the fermented fruit pomace by inoculation of horse feces. For the growth of Bacillus subtilis, Saccharomyces cerevisiae, and Lactobacillus plantarum, they were inoculated in 3-step order at 0, 12, and 24 h, respectively. The fruit pomace was fermented for 48 h at $35^{\circ}C$. The pH of the apple, carrot, grape, citrus and all mixed pomaces decreased from 5.45~6.25 to 4.40~4.77. Microbial growth was maintained at $10^8{\sim}10^9cfu/g$. After 12 and 24 h incubation, dry matter of carrot pomace were highest at 54.84 and 56.66%, respectively (P<0.05) and that of grape pomace was lower than others during fermentation (P<0.05). Dry matter was generally reduced by about 20%. NDF decreased gradually or maintained after 24 h, indicating the fiber degradation. Ash content tended to decrease during fermentation. After 48 hours fermentation, Bacillus, yeast and Lactobacillus showed an excellent growth by using juice by-products. These results suggest that fermented juice pomace has a potential as horse feedstuff with probiotics to maintain beneficial microflora in horse gut.

• Journal of Life Science
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• v.21 no.4
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• pp.509-514
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• 2011

Maloactic fermentation (MLF) occurs after completion of alcoholic fermentation and is mediated by lactic acid bacteria (LAB), mainly Oenococcus oeni. Kiwi wine more than commercial grape wine has the problem of high acidity. Therefore, we investigated the optimal MLF conditions for regulating strong acidity and improving the quality properties of wine fermented with Kiwi fruit cultivated in Korea. For alcohol fermentation, industrial wine yeast Saccharomyces cerevisiae KCCM 12650 strains and LAB, known as MLF strains, were used to alleviate wine acidity. First, the various experimental conditions of Kiwi fruit, initial pH (2.5, 3.5, 4.5), fermenting temperature (20, 25, $30^{\circ}C$), and sugar contents (24 $^{\circ}Brix$), were adjusted, and after the fermentation period, we measured the acidity, pH, and the change in organic acid content by the AOAC method and HPLC analysis. The alcohol content of fermented Kiwi wine was 12.75%. Further, total acidity and pH of Kiwi wine were 0.78% and 3.5, respectively. Total sugar and total polyphenol contents of Kiwi wine were 38.72 mg/ml and 60.18 mg/ml, respectively. With regard to organic acid content, the control contained 0.63 mg/ml of oxalic acid, 2.99 mg/ml of malic acid, and 0.71 mg/ml of lactic acid, whereas MLF wine contained 0.69 mg/ml of oxalic acid, 0.06 mg/ml of malic acid, and 3.12 mg/ml of lactic acid. Kiwi wine had lower malic acid values and total acidity than control after MLF processing. In MLF, the optimum initial pH value and fermentation temperature were 3.5 and $25^{\circ}C$, respectively. Therefore, these studies suggest that establishment of optimal MLF conditions could improve the properties of Kiwi wine manufactured in Korea.

• Applied Biological Chemistry
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• v.20 no.1
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• pp.66-80
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• 1977

This research was carried as a part of the basic study, in which the aptitude of theKorean oak varieties as barrels for aging apple fine spirits was investigated, and thefollowing results were obtained. 1. Following was the result of the chemical analysis of the fruits which are now mass-produced and can be used as a substitute for raw materials for wine production. Apple (Malus pumila Miller var. domestica Schneider) : Total sugar. total acid, volatile acid and pectin of Jonathan (Hong-og) were 13.95%, 0.46%, 0.012%, 0.20% respectively. Total sugar, total acid, volatile acid and pectin of Ralls (Koog-kwang) were 13.35%, 0.43%, 0.011%, 0.45% respectively. 2. Because of low yield of apple juice due to cellulose, pectin, hemicellulose which are present besides sugars, acids in apples, the apple juice were treated with xylanase of Aspergillus niger SUAFM-430, cellulase and pectinase of Aspergillus niger SUAFM-6. This treatment increased the yield of apple juice. And the apple juice was sterilized by adding potassium metabisulfite $(K_2S_20_5)$ and Saccharomyces cerevisae var. ellipsoideus Rasse Johannisberg II (SUAFM-1018) as a cultivation yeast, which has a strong fermentation power was used to ferment. The yield of apple wine based on raw material was 86-87%. The amount of ethanol, extract and methanol obtained from Jonathan and Ralls were 13.5%, 5.4%, 0.04-0.05% respectively. 3. Wines were distilled for two times by the pot still method to make fine spirits. The yield of fine spirits from apple wine mash was 86.6%, and the pH of fine spirits from Jonathan and Ralls were 4.1, 4.2 respectively. 4. The oak chips made of inner part or outer part of 24 Korean oak varieties were used to select the barrel for aging fine spirits. Two oak chips (one oak chip: $1{\times}1{\times}5cm$) of the inner part or of the outer part of each oak variety were dipped into 300 ml of fine spirits, which was bottled in 640ml beer bottle, and followed aging. The colors, flavors and tastes of the fine spirits were checked during 6 months. A. As a criterion for the first screening of oak barrels for aging fine spirits, the rate five of color extraction was determined. The oak chips showed good results in their order as follows and the best 5 varieties were selected. Gal-cham: Quercus aliena Blume (Inner part), Gul-cham: Quercus variabilis Blume (Outer part), Gal-chain: Quercus aliena Blume (Outer part), Jol-cham: Quercus serrata Thumb (Inner and Outer part). Sin-gal-cham: Quercus mongolica Fisher (Outer and Inner part) Sang-su-ri: Quercus acutissima Carruthers (Outer and Inner part) B. To find out the influence of aging temperature on aging, apple fine spirits were aged by dipping each oak chip at room temperature $(24-25^{\circ}C)$) and $45^{\circ}C$. Aging at $45^{\circ}C$ gave the best result followed aging at $30^{\circ}C$ and then at room temperature. C. Apple fine spirits was aged for six months by dipping oak chips in Erlenmeyer flasks and was irradiated with U.V light. The U.V irradiation enhanced the aging effect by nearly two times, compared with the aging without U.V irradiation. D. In aging apple fine spirits by dipping two oak chips, it was observed that the extent of the extraction of most components of oak chips were strongly dependent upon the pH of fine spirits. E. Oak chips of five selected oak varieties and a Limousin white oak from France as a control were used. Each apple fine spitits was dipped by two oak chips, and was aged at room temperature $(24-25^{\circ}C)$, $30^{\circ}C$, $45^{\circ}C$, and with the U.V irradiation at room temperature shaking every week. After six months of aging, the panel test of these aged fine spirits (Young Brandy) showed the following result. Young brandy of apples aged at $45^{\circ}C$ by dipping oak chips of Gal-chain was almost as the fine spirits which were aged at room temperature by dipping Limousin white oak chips from France. Young brandy of with U.V. irradiation at room temperature which were aged by dipping oak chips of Gal-chain was a little worse than that from the fine spirits aged at room temperature by dipping Limousin white oak chips from France. And so, Korean oak varieties are thought to be able to be used for aging every apple fine spirit which was here investigated.