• Journal of the Korean Society of Food Culture
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• v.32 no.6
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• pp.566-575
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• 2017

This study was conducted to screen for superior strains for preparation of Cheonggukjang and identify the optimal fermentation time based on fermentation of lentils by various Bacillus subtilis strains. Bacillus count was significantly high at 48 hrs of fermentation (9.22-9.51 log CFU/g). In addition, the pH was significantly high in the range of 7.60-7.92 at 48 hrs fermentation, and the reducing sugar content was significantly high (0.89-1.45). The amino-type nitrogen content and ammonia-type nitrogen content were significantly increased with fermentation time. Additionally, the amylase activity was significantly high (3.05-4.22) at 48 hrs of fermentation, as was the protease activity (4.80-5.63). Bacillus subtilis S3 (5.63), S4 (5.60) were higher than S1 (4.80), S2 (4.92). The viscous substances and DPPH radical scavenging activity increased at 48 hrs, then decreased. Sensory evaluation revealed that lentil Cheonggukjang fermented by S3 showed the highest scores in color, smell, savory taste, sweet taste, viscous substance and overall acceptability. Therefore, 48 hrs of fermentation should be suitable for lentil Cheonggukjang fermented by Bacillus subtilis S3.

• Korean Journal of Food Science and Technology
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• v.22 no.6
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• pp.611-617
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• 1990

This study was conducted to breed a high vitamin $B_{12}$ producer by the fusion of protoplasts between Bacillus natto and Bacillus megaterium. Auxotrophic mutants of Bacillus natto SH-34 ($thr^-try^-rif^r$) and Bacillus megaterium BK-13 ($arg^-ade^-lys^-str^r$) which showed high protease activity and production of vitamin $B_{12}$, respectively, were isolated for the fusion experiment. Protoplasts were induced by incubating the cells with lysis solution containing $500{\mu}/ml$ lysozyme, and the ratio of protoplast and regeneration formation were ranged from 99% and 67%, respectively. Fusion frequencies of fusants between Bacillus natto SH-34 and Bacillus megaterium BK-13 were appeared in the ranges of $1.0{\times}10^{-5}$ under the treatment of 30% PEG 6000 containing 3% PVP. The fusant, MNF-72 showed the highest product yield of $7.85{\mu}g/g-cell\;vitamin\;B_{12}$ in production medium. For the improvement of productivity, the immobilization of fusants with sodium alginate was carried out. In batch and continuous fermentation systems, the productivity were determined to be $0.58{\mu}g/ml.hr\;and\;0.80{\mu}g/ml.hr\;vitamin\;B_{12}$ under optimum condition, respectivity.

• Preventive Nutrition and Food Science
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• v.11 no.2
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• pp.127-132
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• 2006

Bacillus sp. strain K-l, which produces a strong fibrinolytic enzyme, was isolated from chongkukjang, a traditional Korean fermented soybean paste. The fibrinolytic enzyme was purified from chongkukjang base by using ammonium sulfate fractionation and chromatographic techniques. Purified enzyme, CK K-1 was demonstrated to be homogeneous by SDS-PAGE and isoelectric focusing electrophoresis, and has molecular mass of a 12.4 kDa and a pI of 8.0. The optimal reaction pH value and temperature were 8.0 and $40^{\circ}C$, respectively. Phenyl-methyl-sulfonyl-fluoride (PMSF; serine protease inhibitor), ethylene-diamine-tetra-acetic acid (EDTA; metallo protease inhibitor), copper ion, ferric ion and lead ion inhibited the enzyme activity. These results indicated that the fibrinolytic enzyme is a metallo-serine protease and different from nattokinase and chongkukjangkinase.

• Journal of the Korean Society of Food Science and Nutrition
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• v.26 no.5
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• pp.751-758
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• 1997

Fifty three microbes, mainly fungal genera, were isolated from sixteen Mejus of different region. From those collected isolates, Meju was manufactured and assayed for the activities of amylase and protease. Correlations between sensory evaluation and color measurement were investigated with Kanjang (soy sauce) prepared by each pure inoculation. Color of Kanjang was quite various depedning on fungal genera, but the taste was not quite related with the activity of amylase or protease. This fact might mean that taste of Kanjang depended on the complicate mechanistic action of enzyme for the substrate involved in the soybean hydrolysis. Thus, the taste of Kanjang origenated from Korean traditional Meju seems to belong to complex flora of participated fungal genera as well as Bacillus. sp.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.5
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• pp.702-710
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• 2016

The bioactivity of soy grits cheonggukjang was enhanced by fermentation using three ${\beta}$-glucosidase-producing Bacillus subtilis strains (HJ 18-9, HJ 25-8, and HJ 18-9+HJ 25-8) for 48 h at $37^{\circ}C$. The results indicate that protease, cellulase, and a-amylase activities significantly increased (P<0.05) with increasing fermentation time. In addition, the amino-type nitrogen content of B. subtilis-fermented soy grits cheonggukjang increased to 91.0~168.0 mg% after 48 h of fermentation. Among the isoflavones in soy grits cheonggukjang, contents of ${\beta}$-glucosides or acetyl-glucosides were reduced while aglycone content increased upon fermentation. In particular, soy grits cheonggukjang fermented with B. subtilis HJ18-9 and HJ25-8 showed the largest increases in aglycone content compared to complex treatment. These results provide useful information for development starter (single and complex) as well as for production of high quality fermented soybean food.

• Korean Journal of Food Science and Technology
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• v.41 no.6
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• pp.644-651
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• 2009

This study was conducted to investigate the effects of steeping periods and temperatures of waxy rice on the microflora and enzyme activities of steeping water and waxy rice, as well as on the microscopic structures of waxy rice during the preparation of gangjung. When the steeping water was 15 and $35^{\circ}C$, yeast and lactic acid bacteria were the major microflora after 1 day. After 11 days, Lactobacillus spp. were the major bacteria, while Bacillus spp. were the primary microflora after 21 days. At $25^{\circ}C$ the primary species were Bacillus spp., Leuconostoc spp., and Bacillus spp. after 1, 11 and 21 days, respectively. The $\alpha$-amylase activity of steeping water increased proportionally with the increase of temperature and period, while the protease activity tended to increase for 11 days, but decreased at 21 days. The enzyme activities of the steeped waxy rice, especially the $\alpha$-amylase activity, decreased with higher steeping temperature, while the $\beta$-amylase and glucoamylase activity remained constant for up to 21 days. Observation using scanning electron microscopy indicated that intracellular cleavage of steeped waxy rice endosperm resulted in a rough surface morphology due to the exposure of polygonal starch granules. Tiny pinholes on the surface of waxy rice starch granules were also observed after steeping for 11 days.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.1
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• pp.108-114
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• 2017

This study was performed to evaluate the characterization of fermented mulberry leaf using Bacillus subtilis by focusing on protein degradation and antioxidant activity. The crude protein and total amino acid compositions of mulberry leaf were 21.40% (w/w) and 105.06 mg/g, respectively. The pH level decreased sharply in mulberry leaf extracts fermented using B. subtilis, in accordance with an increase in bacteria cell populations (9.49 log CFU/mL) during 36 h of fermentation. The protease activity of mulberry leaf increased to 97.45 units/mL after 5 days of fermentation. After fermentation, free amino acid contents in fermented mulberry leaf increased from $486.91{\mu}g/g$ to $644.35{\mu}g/g$ due to considerable elevation of isoleucine (6-fold), alanine (4.67-fold), leucine (4.52-fold), and valine (4.21-fold). The DPPH radical scavenging activity of fermented mulberry leaf also increased from 25.93% to 73.22% after 5 days of fermentation. These results suggest that mulberry leaf fermentation using B. subtilis can improve nutritional quality and antioxidant activity.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.5
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• pp.756-762
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• 2014

This study evaluated the quality characteristics of soybean fermented with several Bacillus sp., which were selected based on their high enzymatic and antimicrobial activities. Total aerobic bacterial counts of fermented soybeans with HJ5-2 ($3.00{\times}10^9CFU/mL$) were the highest among all strains. Lactic acid bacteria numbered $2.50{\times}10^2{\sim}7.30{\times}10^4CFU/mL$ in soybeans fermented with isolates. Amylase and protease activities of the RD7-7 sample were the highest among all strains. Reducing sugar and amino-type nitrogen contents of fermented soybeans with HJ18-4 (2.35%) and RD7-7 (227.96 mg%) were the highest. Total amino acid contents of the samples were 16.62~18.38%, and glutamic acid, aspartic acid, leucine, lysine, arginine were major amino acids. Oxalic acid (36.51~63.57 mg/100 g) and succinic acid (429.49~600.15 mg/100 g) were the predominant organic acid. These results provide useful information for development starter (single and complex) as well as for the production of high quality fermented soybean foods.

• Microbiology and Biotechnology Letters
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• v.41 no.1
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• pp.33-43
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• 2013

A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at $37^{\circ}C$ reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of $40^{\circ}C$ and an optimum pH of 8.0, and the enzyme was stable in the ranges $20-40^{\circ}C$ and pH 6.0-8.0. Enzyme activity was increased by $Ca^{2+}$ and $Co^{2+}$ but inhibited by $Cu^{2+}$, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.

• Journal of Life Science
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• v.20 no.3
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• pp.403-410
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• 2010

In order to produce high-quality fermenting composts, a microorganism was isolated from the natural world. The bacterium has not only in high enzyme activities but also had good antimicrobial activities against phytopathogenic microorganisms. Its cultivating characteristics were then investigated. Bacterium KJ-9, which contains high CMCase, protease and chitinase activities and excellent antimicrobial activities against phytopathogenic microorganisms, was separated from leaf mold and identified as Bacillus licheniformis by two methods: Bergey's Manual of Systematic Bacteriology and API 50 CHL Carbohydrate Test Kit (Bio Merieux, France) using an ATB (Automated Identification) computer system (Bio Merieux, France). Optimal medium for cultivation of B. licheniformis was 2% soluble starch as a carbon source, 0.5% yeast extract as a nitrogen source and 0.05% $MgSO_4{\cdot}7H_2O$. Optimal growth conditions of pH, temperature and shake speed were pH 7.0, $50^{\circ}C$ and 180 rpm, respectively. Culture broth of B. licheniformis KJ-9 cultured for 36~60 hr was effective in fungicidal activities against plant pathogens including Botrytis cinerea, Corynespora cassicola, Fusarium oxysporum, and Rhizoctonia solani.