• Journal of the Korean Society of Food Science and Nutrition
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• v.31 no.2
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• pp.343-350
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• 2002

This study was conducted to standardize the manufacturing process of doenjang. The preparation methods, kinds and levels of the ingredients were determined by the statistical surveys of literatures obtained from cooking books, scientific papers and doenjang manufacturing factories. The standardized preparation of fermentation methods of doenjang were classified into two large groups, that were traditional and modified (commercialized) methods. Most soybeans used in doenjang preparation were the large size. To prepare traditional doenjang, soybeans were cleaned, scaled and cooked for 2 hrs at atmospheric pressure. These cooked soybeans were crushed in water and molded as brick shape. The molded soybean was dried for 2 days in the air, hung up by rice straw and fermented for 30~60 days under natural environmental condition (called meju). Recently soybean grain meju that inoculated with Asp. oryzae also frequently used to make traditional doenjang. The fermented meju was brined with a ratio of meju : salt : water = 18.4 : 14.6 : 67.0 and the meju-brine mixtures were ripened for 2 months. When the meju-brine mixture was fully fermented, it was separated into liquid and solid parts. The crushed solid part was further ripened in a separated pottery for 60 days and become doenjang. The liquid part was filtered, boiled and used as soy sauce. In modified commercial doenjang preparation, soybeans were cocked by autoclaving and then cooled about to 3$0^{\circ}C$. Separately, steamed barley grains or wheat flour were inoculated with 0.2% Asp. oryzae and incubated for 3 days at 3$0^{\circ}C$ and mixed with the crooked soybeans, salt, and water (soybean : salt : starch : water = 39.8 : 12.5 : 22.6 : 25.1). These mixtures were ripened for 30 days at 3$0^{\circ}C$. It seems that the manufacturing process of traditional doenjang needs to be more industrialized, whereas, the commercial doenjang preparation is going to adapt the traditional processing method of doenjang.

• The Korean Journal of Mycology
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• v.43 no.3
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• pp.149-157
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• 2015

We examined taxonomic characteristics and safety of eight Nuruk molds that are widely used for making soybean paste, soy sauce and alcoholic beverages in Korea. HK1 from Hakyeong Fermentation Co., SW101 from Suwon Fermentation Co., CF1001, CF1002, CF1003 from Chungmoo Fermaentation Co. and KACC 93210 are yellow-Nuruk molds, and SW201 from Suwon Fermentation Co. and CF1005 from Chungmoo Fermentation Co. are white-Nuruk molds. Six strains of yellow-Nuruk molds were identified as Aspergillus oryzae. HK1, SW101, CF1001 and CF1003 of yellow-Nuruk molds have middle length of stipes ($711{\sim}1,121{\mu}m$), and CF1003 (for sake) produced less conidia and more hyphae than HK1, SW101 and CF1001 (for soybean paste). CF 1002 used for soy sauce has shorter stipes ($543{\mu}m$) and is clustered into IBLB-group based on omtA gene analysis although the other yellow-Nuruk molds are clustered into ICAo group. KACC 93210 isolated from traditional Korean Meju has very short stipes (average $270{\mu}m$), and showed velvety colonies although the others showed floccose colonies. The strain has different DNA sequences of omtA gene from other strains in NCBI GenBank as well as strains used in Korea, suggesting that it is unique from other strains published. SW201 and CF1005 of white-Nuruk molds were identified as Aspergillus luchuensis or A. luchuensis mut. Kawachii that is known as safe, non-toxigenic fungus. The six strains of yellow-Nuruk molds did not produce mycotoxins including aflatoxin, cyclopiazonic acid, and sterigmatocystin. Therefore, eight strains of Nuruk molds used for making soy sauce, soybean paste and alcoholic beverages in Korea were proved to be safe in this study.

• Microbiology and Biotechnology Letters
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• v.41 no.2
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• pp.190-197
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• 2013

Three slime-forming lactic acid bacteria were isolated from traditional Korean fermented soy sauce and soybean paste and shown to produce exopolysaccharides (EPS) in sucrose media. By isolating the strains, examining their morphological characteristics and determining their 16S rDNA sequences, N58-5 and K6-7 were identified as Leuconostoc mesenteroides and N45- 10 as Leuconostoc citreum. The acid and bile tolerances of these three strains were investigated. Amongst the three lactic acid bacteria, Leuc. citreum N45-10 exhibited the highest viability ($10^5-10^6$ CFU/ml) in 0.05 M sodium phosphate buffer (pH 0.3) for 2 h, in artificial gastric juice for 2 h and in 0.3%, 0.5% oxgall for 24h. Leuc. mesenteroides K6-7, N58-5 and Leuc. citreum N45- 10 were grown in sucrose liquid medium and 8.16 g/L, 3.65 g/L, 16.17 g/L of EPS was collected, respectively. The hydrolyzed EPS was analyzed by HPLC in order to determine the sugar composition of EPS. Leuc. mesenteroides K6-7 and N58-5 showed two peaks indicating glucose and fructose, thus they were determined to be hetero-type polysaccharides. Leuc. citreum N45-10 showed only the glucose polymer, indicating it to be a homo-type polysaccharide. In addition, all three lactic acid bacterial hemolysis did not demonstrate a clear zone in blood agar in the area surrounding a lactic acid bacteria colony.

• Journal of Food Hygiene and Safety
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• v.34 no.3
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• pp.296-302
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• 2019

Ganjang-gejang (soy sauce-marinated crab) is a ready-to-eat (RTE) seafood and is also one of the most popular traditional dishes in Korea. It is generally prepared by washing raw blue crabs and then preserving them in soy sauce. Since this process does not involve cooking or any treatment with heat, it is difficult to control the microbiological quality of the final product. Thus, the objectives of this study were to compare the efficacies of various sanitizers in eliminating microorganisms on raw blue crab during the washing step and to evaluate the effectiveness of chitosan on the inhibition of microbial growth in the ganjang-gejang during storage. The raw blue crabs were submerged in chlorinated water (50 mg/L), peracetic acid (40 mg/L), acetic acid (5%) and lactic acid (5%) for 10 min at $25^{\circ}C$, respectively. The blue crabs treated with 5% acetic acid were marinated with soy sauce containing 0.5 and 1% of soluble chitosan, followed by storing them at 4 and $12^{\circ}C$ for up to 30 days. Results show that 5% acetic acid reduced the microbial populations on the blue crabs by 1.5 log CFU/g, which was significantly higher than those of other treatments. Based on these results, 5% acetic acid was selected for the washing step. The microbial populations of all ganjang-gejang samples significantly increased to about 8.0 CFU/g at $12^{\circ}C$ for 7 days. At $4^{\circ}C$, the microbial populations of the products containing 1% chitosan increased by about 2.9 CFU/g for 20 days, which were significantly lower than those (4.2-4.5 log CFU/g) of the products without and with 0.5% chitosan. Thus, these results suggest that 5% acetic acid washing of raw blue crabs and the addition of 1% chitosan in ganjang-gejang could improve the microbiological quality of the final products under refrigerated condition.

• Journal of Life Science
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• v.14 no.3
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• pp.501-508
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• 2004

A bacterial strain CJ-3 which produced chitinase was isolated from Korean traditional soy sauce. Using 16S rDNA analysis, the strain CJ-3 was identified as Bacillus atrophaeus. The approximate molecular weight of the putative chitinase enzyme was 31.0 kDa and the enzyme activity was remarkably induced by addition of colloidal chitin (0.5, 1.0, 2.0%). The antioxidant activity was increased 53% by the browning reaction products of B. atrophneus CJ-3. Escherichia. coli lipopolysaccharides (LPS)-induced production of nitric oxide(NO) was reduced up to 45% by the browning reaction product in RAW264.7 macrophage. Inhibition of cell viability in the presence of LPS was recovered to normal level by the browning reaction product. These results suggest that browning reaction of B. atrophaeus CJ-3 plays an important role for activation of immune system. B. atrophaeus CJ-3 exhibited optimum temperature and pH of 37$^{\circ}C$ and pH 7.0∼8.0, respectively. The major intracelluar free amino acid was determined to be glutamate.

• Preventive Nutrition and Food Science
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• v.16 no.2
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• pp.165-173
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• 2011

Five types of meju were prepared from 100% defatted soybean (DFSG0), a mixture of 90% DFS and 10% glasswort (DFSG1), a mixture of 80% DFS and 20% glasswort (DFSG2), a mixture of 70% DFS and 30% glasswort (DFSG3), and a mixture of 60% DFS and 40% glasswort (DFSG4). Five types of kanjang were separately prepared from the 5 types of meju by ripening in brine for 6 months. The contents of certain minerals (Mg, Ca, Fe, Mn, and Zn), organic acids (citric acid, malic acid) and the antioxidative effects in the kanjang were increased in proportion to the glasswort content in the meju. However, the free amino acid contents in the kanjang were reduced in proportion to the glasswort content in the meju. DFSG1- and DFSG2-kanjang did not show distinct differences from DFSG0-kanjang based on aroma, flavor, and taste that were compared simply by panel tests. The bacterial and fungal community in the fermented meju and kanjang was not affected by the addition of glasswort to the meju-making process. Bacteria belonging to the Lactobacillus and Bacillus genera and the Lactobacillus family predominated, and yeasts belonging to the Saccharomyces genus and fungi belonging to the Aspergillus genus predominated in the fermented meju and kanjang. In conclusion, the glasswort was a supplement that nutritionally improved the kanjang (except for free amino acid contents) but didn't influence the growth of microorganisms that are responsible for the fermentation of meju and kanjang.

• Toxicological Research
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• v.31 no.3
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• pp.299-305
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• 2015

An HPLC analytical method was validated for the quantitative determination of biogenic amines in agricultural products. Four agricultural foods, including apple juice, Juk, corn oil and peanut butter, were selected as food matrices based on their water and fat contents (i.e., non-fatty liquid, non-fatty solid, fatty liquid and fatty solid, respectively). The precision, accuracy, recovery, limit of detection (LOD) and quantification (LOQ) were determined to test the validity of an HPLC procedure for the determination of biogenic amines, including tryptamine, ${\beta}$-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine, in each matrix. The LODs and LOQs for the biogenic amines were within the range of 0.01~0.10 mg/kg and 0.02~0.31 mg/kg, respectively. The relative standard deviation (RSD) of intraday for biogenic amine concentrations ranged from 1.86 to 5.95%, whereas the RSD of interday ranged from 2.08 to 5.96%. Of the matrices spiked with biogenic amines, corn oil with tyramine and Juk with putrescine exhibited the least accuracy of 84.85% and recovery rate of 89.63%, respectively, at the lowest concentration (10 mg/kg). Therefore, the validation results fulfilled AOAC criteria and recommendations. Subsequently, the method was applied to the analysis of biogenic amines in fermented agricultural products for a total dietary survey in Korea. Although the results revealed that Korean traditional soy sauce and Doenjang contained relatively high levels of histamine, the amounts are of no concern if these fermented agricultural products serve as condiments.

• Toxicological Research
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• v.31 no.3
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• pp.289-297
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• 2015

Ethyl carbamate (EC) classified as a probable human carcinogen (Group 2A) is naturally formed in alcoholic beverages and fermented foods during fermentation process and/or during storage. The objective of this study was to analyze EC in 34 food items including 14 alcoholic beverages and 20 fermented foods sold in Korea. Each food was collected from 18 supermarkets in 9 metropolitan cities in Korea, and then made into composite. According to food composition and alcohol content, samples were divided into four matrices such as apple juice, milk, Soju (liquor containing about 20% alcohol), and rice porridge. The maximum EC value of $151.06{\mu}g/kg$ was found in Maesilju (liquor made from Maesil and Soju). Whisky and Bokbunjaju (Korean black raspberry wine) contained $9.90{\mu}g/kg$ and $6.30{\mu}g/kg$, respectively. EC was not detected in other alcoholic beverages. Of 20 fermented foods, Japanese-style soy sauce had highest level of $15.59{\mu}g/kg$ and traditional one contained $4.18{\mu}g/kg$. Soybean paste had $1.18{\mu}g/kg$, however, EC was not found in other fermented foods.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.271-276
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• 2003

Bacillus amyloliquefaciens D4, which produces a strongly fibrinolytic enzyme, was isolated from Chungkook-Jang, a traditional Korean soybean-fermented food. B. amyloliquefaciens D4 was mutated with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) to yield a series of mutants with increasing levels of fibrinolytic enzyme production. After mutation, a mutant D4-7 was obtained with fibrinolytic activity about eight times stronger than the parent strain. The fibrinolytic activity of B. amyloliquefaciens D4-7, reached a maximum, when the producer was cultivated in 2% Isolated Soy Protein (ISP) broth for 48 h at $37^{\circ}C$. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of B. amyloliquefaciens D4-7 showed stronger activity than plasmin and streptokinase. The optimum temperature and pH were $50^{\circ}C$ and 10.0 and thermostability was increased by the addition of glycerol, glucose, and NaCl.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.9-18
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• 2017

Nine bacilli with fibrinolytic activities were isolated from doenjang, a traditional Korean fermented soy food. Among them, RSB34 showed the strongest activity and was identified as Bacillus amyloliquefaciens by 16S rRNA and recA gene sequencing. During growth on LB up to 96 h, RSB34 showed the highest fibrinolytic activity ($83.23mU/{\mu}l$) at 48 h. Three bands of 23, 27, and 42 kDa in size were observed when the culture supernatant was analyzed by SDS-PAGE and 27 and 42 kDa bands by fibrin zymography. The gene encoding the 27 kDa fibrinolytic enzyme AprE34 was cloned by PCR. BLAST analyses confirmed that the gene was a homolog to genes encoding AprE-type proteases. aprE34 was overexpressed in Escherichia coli BL21(DE3) using pET26b(+). Recombinant AprE34 was purified and examined for its properties. The $K_m$ and $V_{max}$ values of recombinant AprE34 were $0.131{\pm}0.026mM$ and $16.551{\pm}0.316{\mu}M/l/min$, respectively, when measured using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. aprE34 was overexpressed in B. subtilis WB600 using pHY300PLK. B. subtilis transformants harboring pHYRSB34 (pHY300PLK with aprE34) showed higher fibrinolytic activity than B. amyloliquefaciens RSB34.