• Mycobiology
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• v.44 no.3
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• pp.155-161
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• 2016

The most economically important species used in a wide range of fermentation industries throughout Asia belong to Aspergillus section Flavi, which are morphologically and phylogenetically indistinguishable, with a few being toxigenic and therefore a major concern. They are frequently isolated from Korean fermentation starters, such as nuruk and meju. The growing popularity of traditional Korean alcoholic beverages has led to a demand for their quality enhancement, therefore requiring selection of efficient non-toxigenic strains to assist effective fermentation. This study was performed to classify the most efficient strains of Aspergillus section Flavi isolated from various types of traditional wheat nuruk, based on a polyphasic approach involving molecular and biochemical evaluation. A total of 69 strains were isolated based on colony morphology and identified as Aspergillus oryzae/flavus based on internal transcribed spacer and calmodulin gene sequencing. Interestingly, none were toxigenic based on PCR amplification of intergenic regions of the aflatoxin cluster genes norB-cypA and the absence of aflatoxin in the culture supernatants by thin-layer chromatography analysis. Saccharification capability of the isolates, assessed through ${\alpha}-amylase$ and glucoamylase activities, revealed that two isolates, TNA24 and TNA15, showed the highest levels of activity. Although the degrees of variation in ${\alpha}-amylase$ and glucoamylase activities among the isolates were higher, there were only slight differences in acid protease activity among the isolates with two, TNA28 and TNA36, showing the highest activities. Furthermore, statistical analyses showed that ${\alpha}-amylase$ activity was positively correlated with glucoamylase activity (p < 0.001), and therefore screening for either was sufficient to predict the saccharifying capacity of the Aspergillus strain.

• Journal of the Korean Society of Food Culture
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• v.11 no.3
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• pp.339-348
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• 1996

Physicochemical and sensory characteristics of Korean traditional alcoholic beverages, such as Kwahaju, Sokokju, Baikhaju, Samhaeju and Hosanchun were investigated during fermentation with the difference of input step and treatment of rice and Nuruk (Korean-style bran koji). pH values in all treatments were similar to be ranged $3.1{\sim}3.2$, but acidity of the treatment brewed without seed mash was $1.5{\sim}2$ times higher than that of the treatment brewed with seed mash. The concentration of alcohol in all treatments increased as the concentration of the Nuruk increased. The alcohol concentrations of Samhaeju and Hosanchun ranging from 6.5% to 7.7% were higher than those of Kwahaju, Sokokju and Baikhaju ranging from 7.9% to 10.6%. Residual reducing sugar contents of Kwahaju, Sokokju and Baikhaju ranging from 4.6 g/L to 9.7 g/L were lower than those of Samhaeju and Hosanchun ranging from 14.2 g/L to 15.5 g/L. The consumed total sugar contents of Kwahaju, Sokokju and Baikhaju ranging from 125.4 g/L to 159.2 g/L were higher than those of Samhaeju and Hosanchun ranging from 111.2 g/L to 123.8 g/L, and the treatments brewed with seed mash showed more sugar consumption than the treatments brewed without seed mash. Sensory quality in the treatments brewed with seed mash was better than those of without seed mash. Concentration of Nuruk and preparation of seed mash from it were critical factors compared to input step and treatment of rice in Korean traditional alcoholic beverage brewing.

• The Korean Journal of Food And Nutrition
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• v.36 no.5
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• pp.360-367
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• 2023

The major active components of Astragalus membranaceus (AM) are isoflavones, which exist in the form of various glycosides. Nuruk is a traditional fermentation starter in Korea, and is suitable for the biotransformation of isoflavone glycosides because it contains various microorganisms and enzymes. This study was performed to determine changes in the isoflavones and antioxidant properties of AM fermented (AF) with nuruk over 24 hours. AF was sampled after 0, 2, 4, 6, 12, 18, and 24 h of fermentation, and calycosin 7-glucoside, ononin, calycosin, and formononetin content, and the antioxidant properties of AF were analyzed. The total phenolic content increased with fermentation time, and the ABTS radical scavenging activity increased until 6 h of fermentation and then decreased. During fermentation, the isoflavone glycosides decreased significantly as fermentation time increased. The contents of calycosin and formononetin, which are aglycons of calycosin-7-glucoside and ononin, increased from 100.54 ㎍/g to 276.84 ㎍/g and from 56.29 ㎍/g to 123.04 ㎍/g, respectively, at 18 h of fermentation. Significant correlations were observed between fermentation time, isoflavone content, and antioxidant properties. The results of this study showed that fermentation with nuruk is suitable for the biotransformation of isoflavones in AM.

• Journal of the East Asian Society of Dietary Life
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• v.23 no.5
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• pp.620-628
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• 2013

In this study, the condition for preventing abnormal fermentation was set by heating the nuruk extract, such that glycosyl enzymes maintain its activity and unnecessary microbes are removed. The total colony of microbes in the heated nuruk extract was highest in number at $25^{\circ}C$ and began to reduce at 50, $60^{\circ}C$ and sharply reduced over $70^{\circ}C$. Saccharogenic power (SP), glucoamylase and acidic protease activities were highest at $50^{\circ}C$, 10, 20, 30min and ${\alpha}$-amylase activity was lower at $50^{\circ}C$ than at $25^{\circ}C$. In the pHs of the nuruk extract, as the heat temperature became higher and treatment time was longer, the pHs were reduced significantly. The total acidities of heat treatments at 50, $60^{\circ}C$ were lower by 0.2% than at $25^{\circ}C$, where as the 70, $80^{\circ}C$ treatments showed a sharp rise from the early stage of fermentation. Soluble solids showed the same aspects with the glycosyl enzymes cases. In reducing sugar, 25, $50^{\circ}C$ treatments were sharply increased from the first day of fermentation while $60^{\circ}C$ treatments began to rise from second day 70, $80^{\circ}C$ were slightly increased after the fourth day. The normally alcohol fermented treatments were 25, 50 and $60^{\circ}C$ 30min. The $70^{\circ}C$ treatments almost did not alcohol fermentation. In the preference tests, taste and total acceptability were high at 25, $50^{\circ}C$ treatments. These results suggest that makgeolli using heat treated nuruk extract also has good taste as well as did not.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.11-11
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• 2015

A total of sixty-six samples of Nuruk, a fermention starter used to make the Korean traditional rice wine, Makgeolli, were collected from central and southern regions of Korea in 2013 and 2014. We classified two groups of the Nuruk samples, "commercial" and "home-made", according to the manufacturing procedure and purpose of use. Commercial Nuruks were made in a controlled environment where the temperature and humidity are fixed and the final product is supplied to Makgeolli manufacturers. Home-made Nuruks were made under uncontrolled conditions in the naturally opened environment and were intended for use in the production of small amounts of home-brewed Makgeolli. We obtained more than five hundred isolates including filamentous fungi and yeasts from the Nuruk samples followed by identification of fungal species. Also we stored glycerol stocks of each single isolate at $-70^{\circ}C$. We identified the species of each isolate based on the sequences of ITS regions amplified with two different universal primer pairs. We also performed morphological characterization of the filamentous fungi and yeast species through observations under the microscope. We investigated the major fungal species of commercial and home-made Nuruks by counting the colony forming units (CFU) and analyzing the occurrence tendency of fungal species. While commercial Nuruks contained mostly high CFU of yeasts, home-made Nuruks showed relatively high occurrence of filamentous fungi. One of the representative Nuruk manufacturers used both domestic wheat bran and imported ones, mainly from US, as raw material. Depending on the source of ingredient, the fungal diversity was somewhat different. Another commercial Nuruk sample was collected twice, once in 2013 and again in 2014, and showed different diversity of fungal species in each year. Nuruks obtained from the southern regions of Korea and Jeju island showed high frequency of yeast such as Saccharomycopsis fibuligera and Pichia species as well as unique filamentous fungus, Monascus species. S. fibuligera was easily found in many Nuruk samples with high CFU. The major filamentous fungi were Aspergillus, Lichtheimia, Mucor and Penicillium species. In order to further our understanding of the isolates and their potential industrial applications, we assayed three enzymes, alpha amylase, glucoamylase and acid protease from 140 isolates out of about five hundred isolates and selected about 10 excellent strains with high enzyme activities. With these fungal isolates, we will perform omics analyses including genomics, transcriptomics, metabolic pathway analyses, and metabolomics followed by whole genome sequencing of unique isolates associated with the basic research of Nuruk and that also has applications in the Makgeolli making process.

• Journal of the East Asian Society of Dietary Life
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• v.22 no.1
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• pp.103-108
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• 2012

Nuruk is a fermented ingredient used for production of traditional Korean rice wine. In this study, quality characteristics of mixed-grain nuruk was made by brewing with different fungal strains was analyzed. Quality elements including enzyme activity and organic acids constituents were measured. The fermentation time of the nuruk did not make a significant difference in terms of its pH, but the acidity and amino acid content for nuruk made from a mixture of two fungal strains was higher than that seen with a single fungal strain. Overall, the enzyme activity for two fungal strain nuruk was higher than that observed for single fungal strain nuruk, with ${\alpha}$-amylase and acidic protease activity in the mixed strain nuruk observed to be more than twice that of the single strain. The major organic acids observed in the manufactured nuruk were identified as acetic, citric, formic, fumaric, lactic, malic and oxalic acids. The total amount of organic acids contained in the nuruk made with the two fungal strain was (2,116.3 mg%). The fungal strains used were A. kawachii SC60 nuruk (1,608.5 mg%) and A. oryzae RIB1353 nuruk (1,146.7 mg%).

• Korean journal of food and cookery science
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• v.9 no.4
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• pp.266-271
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• 1993

In order to investigate the traditional value of Ewhaju (traditional wine) and to establish the brewing condition, studies on of traditional background and field inquiry were carried out. Scientific evaluation and possibility of revealation of Ewhaju were searched by the experiments of microbial and enzymatic properties of brewed Ewhaju and Nuruk by traditional method. In flora of microorganisms in Nuruk of Ewhaju, Aspergillus oryzae and Hansenula sP. were isolated, and, showed a level of 1.2$\times$$10^6$ CHU/g, respectiveln but other microorganisms were not grown in diluted cultivation test. The a-and f-amylase activity of Nuruk were 30.74 and 34.4, respectively and their activities of two amylases were 19.28 and 18.8 at first stage of brewing, 21.21 and 19.80 at 100 day after brewing, and 20.25 and 19.90 at one year aged Ewhaju, respectively. The brewed Ewhaju could be remained with high quality long period without teat treatment or addition of preservatives, also, stored Ewhaju contains remarka-bly high activity of amylases which might contribute to digestion.

• Journal of the FoodService Safety
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• v.5 no.1
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• pp.30-39
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• 2024

Rice, nuruk, and water are the main ingredients to prepare takju, a traditional Korean liquor. Takju quality varies depending on the type of ingredients and fermentation temperature. In the current study, different combinations of nuruk (SH, JJ) and rice (glutinous rice, non-glutinous rice) were fermented at different temperatures (20, 25℃) and analyzed for quality characteristics over the fermentation period of 10 days. Regardless of the temperature, higher soluble solids (27.1~29%) were measured in the takju prepared with glutinous rice compared to non-glutinous rice. And acidity increased from 0.03% to 1.1% to all takju fermented at both 20 and 25℃, and, higher acidity was measured in the takju prepared with JJ nuruk. The color value of lightness (L) decreased from 57.9 to 23.9. Alcohol content was less than 15.5% in glutinous rice takju and less than 12.3% in non-glutinous rice takju. Total bacteria count (7~8 Log CFU/mL) and lactic acid bacteria count (4~7 Log CFU/mL) increased in different takju. However, the very high yeast count was recorded in takju prepared from SH nuruk compared to takju prepared from JJ nuruk at 0 day of fermentation. A sharp decline (84.92~98.57%) of yeast was observed just after the 1st day of fermentation which gradually reduced to lowest at 4th day of fermentation. No yeast was observed at 6th day fermentation and onwards. These results indicate that the raw materials, especially rice and nuruk, affect the physiochemical properties of takju.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.11
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• pp.376-385
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• 2019

This study was undertaken is to isolate brewing yeast suitable for rice beer fermentation from the traditional Nuruk, and to identify the brewing ability of the isolated yeast. After 6 months of research, four brewing yeast isolated from traditional Nuruk showed a normal fermentation pattern in terms of physicochemical data (pH, brix, alcohol content) and higher vitality, as compared to commercial brewing yeast. The concentrations of higher alcohol and ester, that impart the aroma to beer, were 78.4 to 106.5 ppm and 15.1 to 29.3 ppm, respectively. In particular, S. cerevisiae (KCCM 90313) bestowed significantly higher contents of higher alcohol and ester concentrations than rice beer prepared from commercial yeast. We conclude that the four variants of yeast isolated from traditional Nuruk are potentially suitable for manufacturing rice beer. Especially, the S. cevisiae (KCCM 90313) yeast shows excellent yeast activity and aroma production, thereby displaying potential application for manufacturing rice beer in the future.

• Journal of Microbiology
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• v.37 no.2
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• pp.80-85
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• 1999

The purification system of glucoamylase (glucan 1,4-${\alpha}$-glucosidase, EC 3. 2. 1. 3), some characteristics of the purified enzyme and hydrolysis rate of various raw starch were investigated through several experiments. The enzyme was produced on a solid, uncooked wheat bran medium of Aspergillus oryzae NR 3-6 isolated from traditional Korean Nuruk. The enzyme was homogeneously purified 6.8-fold with an overall yield of 28.3% by the criteria of disc- and SDS-polyacrylamide gel electrophoresis. The molecular weight was estimated to be 48 kDa by SDS-PAGE. The optimum temperature and pH were 55$^{\circ}C$ and 4.0, respectively. The enzyme was stable at a pH range of 3.0∼10.0 and below 45$^{\circ}C$. Enzyme activity was inhibited about 27% by 1mM Hg2+. The hydrolysis rate of raw wheat starch was shown to be 17.5-fold faster than the hydrolysis rate of soluble starch. The purified enzyme was identified as glucoamylase because the product of soluble starch by the purified enzyme was mainly glucose by thin layer chromatography.