• Journal of Microbiology and Biotechnology
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• v.30 no.8
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• pp.1207-1213
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• 2020

Aflatoxin B1 (AFB1) is a mycotoxin produced by Aspergillus flavus (A. flavus). AFB1 is reported to have high thermal stability and is not decomposed by heat treatment during food processing. Therefore, in this study, knowing that AFB1 is metabolized by cytochrome P450 (CYP), our aim was to develop a method to detoxify A. flavus-contaminated maize, under normal temperature and pressure, using Escherichia coli expressing human CYP3A4. First, the metabolic activity of AFB1 by recombinant human CYP3A4 was evaluated. As a result, we confirmed that recombinant human CYP3A4 metabolizes 98% of AFB1. Next, we found that aflatoxin Q1, a metabolite of AFB1 was no longer mutagenic. Furthermore, we revealed that about 50% of the AFB1 metabolic activity can be maintained for 3 months when E. coli expressing human CYP3A4 is freeze-dried in the presence of trehalose. Finally, we found that 80% of AFB1 in A. flavus-contaminated maize was metabolized by E. coli expressing human CYP3A4 in the presence of surfactant triton X-405 at a final concentration of 10% (v/v). From these results, we conclude that AFB1 in A. flavus-contaminated maize can be detoxified under normal temperature and pressure by using E. coli expressing human CYP3A4.

• Applied Biological Chemistry
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• v.15 no.3
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• pp.213-219
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• 1972

In order to utilize sweet potatoes for the material of Takju, brewing experiments with raw sweet potatoes, sweet potato chips powder and its koji were conducted; and various tests were carried out on effect of the treatments of acid, alkali, polyphenol oxidase inhibitor, oxidizing and reducing agents upon the prevention against coloring of sweet potato chips by steaming, and on peeling effect of sweet potatoes by the alkali and heat treatments. The results obtained were as follows. 1) In the case of brewing with raw sweet potatose, each plot showed low acid and ethanol content, and its finished Takju had an undersirable color and odor. The plots which were mashed after peeling showed higher ethanol contents than the plots mashed without peeling. 2) In the case of brewing with sweet potato chips powder, each plot contained considerably more amount of ethanol than the plots brewed with raw sweet potatoes, white it contained less amount of acid. The ethanol contents of the plots using wheat bran koji were $10.5{\sim}11.4$ per cent 4 days after mashing, and were higher than those of the plots using malts powder. Their finished Takju was inferior in quality because of the lack of acid and being darkened gradually in process of time. 3) The kojies which were made of sweet potato chips powder with Neurospora sitophila or Aspergillus oryzae had good appearance, but the Takju mashes brewed with these contained remarkably less amount of ethanol. 4) Effect of the treatments of acid, alkali, polyphenol oxidase inhibitor and organic solvents such as ether and ethanol upon the prevention against coloring of sweet potato chips was not recognized. Alum and burnt alum were effective a little on the decolorization, and among the oxidizing and reducing agents tested, potassium permanganate was most effective. 5) Darkening of sweet potato chips powder in course of heating after mixing with water was not affected by pectin and amino acids, but by tannin. 6) Sweet potatoes were not peeled easily by friction after soaking in the boiling solution of 3 per cent alkali for 6 minutes and peeled in boiling water for 12 minutes. From the viewpoint of the results above mentioned, it seems to be necessary to study further on the isolation of microorganisms which are able to decompose the coloring substances and yeasts which are adequate for the fermentation of sweet potatoes in order to utilize sweet potatoes for Takju brewing, because brewing with raw sweet potatoes, sweet potato chips powder and its koji was unsuccessful, and effect of the various treatments on the decolorization of sweet potatoes was not recognized.

• Applied Biological Chemistry
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• v.22 no.3
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• pp.160-165
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• 1979

This study was carried out to investigate effects of iron content on the quality of soy sauce, bean paste and red pepper paste, and to elucidate the origin of iron and change of the contents during production processes. For the first step, the iron contents in commercial soy sauce and changes of the contents during brewing process were determined. The results obtained were as follows. 1, Iron contents of raw materials were 108 ppm in soy bean, 133ppm in defatted soy bean, 79 ppm in wheat, 5 ppm in sodium chloride, 58 ppm in seed koji, 300-2000 ppm in spore of Aspergillus oryzae, 240 ppm in wheat gluten, 20 ppm in sodium carbonate (above figures were of dry weight basis), 6 ppm in hydrochloric acid, 18 ppm in caramel and 0.3ppm in brewing water respectively. 2, Iron contents in koji were 200-240 ppm (as dry weight basis) and increased, more or less, in progress of koji-making period. 3. Iron contents in the mashes during fermentation were 40 rpm after 1 month, 43-47 ppm after 3 months and 49-62ppm after 6 months. 4. In chemical soy sauce, the iron content was 159 ppm after hydrolysis of wheat gluten with hydrochloric acid, and 184 ppm after neutralization. 5. Higher iron contents were detected both in fermented and chemical soy sauce when the concentration of total nitrogen increased, but the levels were higher in chemical soy sauce than in fermented one at the same concentration of total nitrogen. 6. In the case of fermented soy sauce, the iron content in the filtrate was decreased by press-filtration, but no significant change was found between before and after heat-sterilization. 7. Iron contents in commercial soy sauce were varied with the producers, however, the average value was 62.7 ppm as calculated as 1.0 percent of total nitrogen. And the average level of iron in home-made soy sauce produced by conventional method was 37.68 ppm.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.3
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• pp.390-396
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• 1997

Meju is a basis for manufacturing Kanjang, Meju was traditionally prepared at home by different types of process depending on the regional area. It is necessary to standardize and simplify the process of Meju-preparation for Kanjang of good quality. For these purposes, the process of Meju and Kangjang making as well as analysis of commercial Kanjang, were compared. Generally, traditional Meju was prepared by steeping and dehulling the whole soybean. After steeping for 24hr. soybean absorbed water up to 110~120% of its weight. The soaked soybeans were steamed for 2hr. and cooled to 5$0^{\circ}C$. Cooked soybeans were crushed down to the size of 10~15 mesh and molded. Molded soybeans were dried for 2 days in the air, hung up by rice straw and fermented for 20~30 days under natural environmental condition. On the other hand, commercial soybean koji was made of defatted soybean. Defatted soybeans were steeped in water and steamed for 15~30min at 0.7~1.2 kg/$\textrm{cm}^2$. Steamed and defatted soybean was cooled to 4$0^{\circ}C$. Separately, wheat power was roasted at 200~30$0^{\circ}C$ by wheat roaster. Mixture of steamed defatted soybean and roasted wheat powder (5/5 to 7/3) were inoculated with 0.1~0.2% Aspergillus sojae and incubated for 2 days at 3$0^{\circ}C$ with occasional stirring. Chemical analysis showed that traditional soy sauces contained the following composition: NaCl, 20.12~25.42%; total nitrogen, 0.64~0.91%; pure extract, 9.47~11.20%; color, 2.34~4.01; pH, 4.92~5.12. Commercial products contained: NaCl, 15.20~17.19%; total nitrogen, 1.25~1.40%; pure extract, 18.17~21.47%; color, 5.41~21.12; pH, 4.51~4.66 and ethalnol. 2.97~3.12%. Organoleptic test on taste, color and flavor of traditional and commercial soysauce indicated that most of the consumers prefer commercial products to traditional products. Preferrable formulation of Kanjang based on organoleptic test of soy sauces was assumed as containing; NaCl, 16.0%; total nitrogen, 1.40%; pure extract, 19.97%; color, 12.98; pH, 4.61 and ethanol, 2.96.

• Journal of Life Science
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• v.29 no.7
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• pp.755-761
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• 2019

Kojic acid (KA) is produced by Aspergillus oryzae-sort of like mushrooms, which is commonly called as koji in Japan. KA is used as a chelation agent and a preservative preventing oxidative browning of fruits. KA also shows antibacterial and antifungal properties. Because KA stops the production of melanin by inhibiting tyrosinase in the biosynthetic pathway from tyrosine to melanin in skin, it has been applied as a skin lightening ingredient in cosmetics. Since some animal studies have shown that high amounts of KA had side effects such as in liver, kidney, reproductive, cardiovascular, gastrointestinal, respiratory, brain, and nervous system, more efficient KA derivatives are needed to be developed in order to safely apply as a skin lightening ingredient. A series of KA derivatives via conjugated with triazole by click reaction were synthesized and their in vitro tyrosinase inhibitory activities were evaluated. Most of all KA derivatives have shown in moderate tyrosinase inhibitory activities. In case of KA-hybrid compound, 1~3 have shown tyrosinase inhibitory activities about 50~10,000 times more effective tyrosinase inhibitor compared to KA itself. Specifically, the $IC_{50}$ value of KA-hybrid compound, 2 was $0.0044{\pm}0.74{\mu}M$ against tyrosinase. It is about 10,000 times more effective tyrosinase inhibitor compared to KA itself ($IC_{50}=45.2{\pm}4.6{\mu}M$).

• Food Science and Preservation
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• v.21 no.6
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• pp.866-877
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• 2014

The aim of this study was to develop a new functional traditional fermented soybean food using Cudrania tricuspidata fruits and fermentation microbes isolated from traditional fermented food. Aspergillus oryzae koji, Lactobacillus sp., and Bacillus sp. were used for the selection of a suitable microbe for the fermentation of Cudrania tricuspidata fruits, and as a result, Bacillus licheniformis SCDB 1234 was selected. SCDB 1234 enhanced the concentration of kaempferol in the Cudrania tricuspidata fruits from 9.54 to $217.04{\mu}g/g$ (about 22 times). The DPPH radical scavenging activity of the fermented materials was similar to that of BHA and BHT (92~99 ppm). The tyrosinase inhibitory activity was high with arbutin (95 ppm) and kojic acid (90 ppm). Doenjang-added fermentation materials of the Cudrania tricuspidata fruits were developed, and the organic acid, reducing sugar, and free amino acid of the developed Doenjang were analyzed. The pancreatic lipase inhibitory (PLI) activity and ${\alpha}$-glucosidase inhibitory (AGI) activity of the fermentation materials of the Cudrania tricuspidata fruits and the developed Doenjang were investigated, and it was found that after fermentation, the PLI and AGI activities of the fermentation materials of the Cudrania tricuspidata fruits were higher than those before fermentation, and that the AGI activity of the developed Doenjang after aging ($91.25{\pm}0.04%$) was higher than that before aging ($84.89{\pm}0.08%$).

• 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.