• Korean Journal of Food Science and Technology
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• v.39 no.3
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• pp.315-322
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• 2007

This study was carried out to investigate the functional activities of microorganisms isolated from kochujang, such as fibrinolytic, immunostimulating, and cytotoxical activities, and to apply these microorganisms to kochujang products. Ninety-one microbial strains with proteolytic activity were selected from 294 strains isolated from traditional and commercial kochujang. Three strains (TPP 0014, TPP 6013, and TPP 6015) with high fibrinolytic activity were tested for their immunostimulating and cytotoxical activities. For the assessment of macrophage activation, cytokines such as tumor necrosis factor, $interleukin-1{\alpha}$ and nitrogen oxide were measured with the murine macrophage cell line RAW 264.7. In addition, the cytotoxical activities of the three strains were examined by MTT assay on the colon cancer cell line SNU-C4 and normal cell line CHO-K1. Using an API identifying kit, two of the microbial strains (TPP 0014 and TPP 6015) were identified as Bacillus stearothermophilus and the other strain (TPP 6013) was identified as B. amyloliquefacience.

• Food Science of Animal Resources
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• v.28 no.2
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• pp.204-210
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• 2008

A total of 70 colonies were isolated from the Korean human milk samples on the BCP plate count agar. These LAB isolates were subcultured in 10% reconstituted skim milk, and two strain thereof were finally selected for their highest acid productions. These strains were identified as Enterococcusfaecium based on 16S rDNA sequencing data, named as Enterococcus faecium KHM-11. Sugar utilization of E. faecium KHM-11 was investigated by API 50CH kit, and 19 different sugars including D-arabinose, L-arabinose, galactose, D-glucose, D-fructose, and D-mannose were utilized. For fermentation profiles, a yogurt inoculated by E. faecium KHM-11 after 15 hour, reached at pH 4.09, titratable acidity at 1.10% and average viable counts $1.30{\times}10^9\;CFU/mL$. Effects of the administration of yogurt 0.5% of piglet diet to piglets were investigated for growth rate, analysis of blood and incidence of diarrhea. 24 heads of piglets were divided into two groups: the experimental and the control of 12 animals each. The average growth rate in the yogurt-fed group was higher for 21.67%, compared with control (p<0.05). There were no differences in the concentrations of blood glucose, cholesterol, albumin and globulin between the two treatments. Incidence of diarrhea was no in pigs fed yogurt as compare to control.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.249-255
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• 2004

Chinese cabbage ('Baechu') Kimchi was fermented at the three different temperatures right after it was prepared. Samples were taken everyday for measuring bacterial populations, pH, and titratable acidity through the whole periods of fermentation up to 50 days. pH values and developed acidity were significantly affected by the fermenting temperatures of 4, 10, and $20^{\circ}C$, suggesting that different bacterial flora has been established by the temperatures exposed. The modified MRS agar containing vancomycin (300 $\mu$g/mL) was used for isolating the vancomycin-resistant LAB strains and 127 isolates were finally obtained. Of the LAB isolates, 13 isolates were subjected to the identification experiments based on the biochemical characteristics and the molecular-typing approach, an ITS-PCR, whether they belong to the genus Leuconostoc or not. The data obtained from API 50 CHL kit resulted that six isolates were identified as the members of Leuconostoc and six as Lactobacillus brevis strains except for a single isolate YKI 30-0401, which was not able to be identified because its biochemical traits were not matched to the database of API 50 CHL kit. It was noted that some isolates were distinct in a couple of some biochemical characteristics compared with those of the reference Leuconostoc species. To overcome the limitations experienced in the commercial identification products above, an ITS-PCR experiment was also conducted for the isolates, resulting that eight isolates belong to Leu. mesenteroides ssp. mesenteroides or dextranicum with a single band of 564 bp, and four to L. brevis strains. The ITS-PCR profiles clearly differentiated the closely-related LAB isolates for which same results were obtained by the biochemical method. This molecular approach, however, failed to produce the amplicons for the YKI 20-1003, leaving the strain unidentified. Judging from the identification data obtained in the Kimchi fermented at $4^{\circ}C$ or $10^{\circ}C$, Leuconostoc spp. including Leu. mesenteroides/dextranicum were likely predominant species in the earlier stage and L. brevis occurred at the high level through the whole period. By contrast, L. brevis, as one of the major flora, possibly lead the fermentation from the beginning in the Kimchi fermented at $20^{\circ}C$.}C$.

• Journal of Applied Biological Chemistry
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• v.53 no.1
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• pp.51-55
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• 2010

A lactic acid bacterial strain showing fast growth and high acid production in Korean pear puree was isolated from Kimchi. This strain was analyzed by API 50 CHL kit and 16S rRNA sequencing analysis and identified as Leuconostoc (Ln.) mesenteroides KACC 91495P. Korean pear puree was fermented using Ln. mesenteroides KACC 91495P strain at $30^{\circ}C$ for 18 h. The changes of pH, titratable acidity and viable cell number during fermentation were investigated. The pH and titratable acidity were reached to pH 3.86 and 1.09% after 18 h fermentation, respectively. The viable cell population of Ln. mesenteroides KACC 91495P was rapidly increased to $2.0{\times}10^9\;CFU/g$ during the 9 h of cultivation. The contents of lactic acid, acetic acid and malic acid were determined to be 0.213, 0.259, and 0.217% after 18 h fermentation, respectively. The content of polyphenolic compounds, known as antioxidants, in pear puree were enhanced by Ln. mesenteroides KACC 91495P cultivation. The level of total polyphenolic compounds was increased to around 140% of initial concentration. When the fermented pear puree was kept at $4^{\circ}C$, pH, titratable acidity and number of viable cells population were nearly maintained for 13 days.

• Microbiology and Biotechnology Letters
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• v.34 no.1
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• pp.40-46
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• 2006

A bacterial strain producing high level of a phytase was isolated from cattle feces and identified as Bacillus subtilis, and designated as Bacillus sp. CF 5-26. The production of the phytase from Bacillus sp. CF 5-26 reached the highest level after 72 hours at $37^{\circ}C$. The optimum condition of the media for the production of phytase was 10% rice bran extract, 0.1% whey protein powder, $0.01%\;CaCl_{2},\;0.01%\;KH_{2}PO_4$. The phytase was purified 20.3 folds with ethanol precipitation, Sephadex G-100, CM Sepharose CL-6B and Sephacryl S-100-HR column chromatography. The molecular weight of the purified enzyme was estimated to be 66 kDa on SDS-polyacrylamide gel electrophoresis. The purified phytase activity was stable up pH 5.0, 7.0, 11.0 and the remaining activity was 50% when it was treated at $100^{\circ}C$ for 1 hour. The substrate specificity of phytase was most active against sodium phytate and inositol polyphosphate compound. And the phytase hydrolysed tripolyphosphate and pyrophosphate a little. The Km value for the sodium phytate was 0.64 mM and the Vmax value was $4.41\;{\mu}mol/min$.

• Applied Biological Chemistry
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• v.46 no.3
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• pp.176-182
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• 2003

An alkaline fibrinolytic protease-producing bacteria was isolated front Korean traditional soy sauce and identified as Bacillus subtilis K7 from the results of analyses of its morphological and physiological properties, $API^{\circledR}$, and Biolog system. The enzyme was purified by 75% ammonium sulfate fractionation, QAE-Sephadex anion and SP-Sephadex cation exchange column chromatography and Sephadex G-100 gel filtration. The specific activity of the purified enByme was 233.9 unit/mg protein and the yield of enzyme was 3.8%. The homogeneity of the purified enzyme was confirmed by polyacrylamide gel electrophoresis. Molecular mass of the enzyme was estimated about 21,500 Da by SDS-polyacrylamide get electrophoresis and gel chromatography. The optimum temperature and pH for the enzyme activity were $40^{\circ}C$ and 9.0, respectively. The enzyme was stable in a pH range of 5.0 to 12.0, and 60% of its activity was lost on heat treatment at $50^{\circ}C$ for 20 min. The activity of the purified enzyme was inhibited by the presence of $Fe^{2+},\;Ag^{2+},\;Cu6{2+}$, iodoacetate, ethylene diamine tetraacetic acid (EDTA), and trans-1,2-diaminocycloheane-N,N,N',N'-tetraacetic acid (CDTA). The results indicates that the enzyme requires a metal ion for its enzymatic activity.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.355-360
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• 2010

Vibrio parahaemolyticus (V. parahaemolyticus), which is commonly found in raw seafood, causes gastroenteritis in humans. Rapid and effective methods have been developed as culture methods require up to 5-7 days. In this study, real-time PCR was compared with the standard culture method for detecting V. parahaemolyticus in seafood and radish sprout samples. Five hundred grams of the samples were artificially contaminated with V. parahaemolyticus then divided into 20 samples. The samples were incubated in alkaline peptone water and then streaked onto thiosulfate-citrate-bile saltssucrose agar. Biochemical tests for suspicious colonies were performed using the API 20NE strip. In parallel, real-time PCR was performed targeting the toxR gene using the enrichment broth. The real-time PCR was sensitive in discriminating V. parahaemolyticus from other foodborne pathogens. The detection limit of the real-time PCR was $10^3\;CFU/mL$ in phosphate-buffered saline. Although the real-time PCR detected more positive samples (76 out of 180, 42%) than the culture method (66 out of 180, 37%), there was no significant statistical difference (p>0.05) between the two methods. In conclusion, real-time PCR assays could be an alternative to the standard culture method for detecting V. parahaemolyticus in seafood and radish sprouts, which has many advantages in terms of detection time, labor, and sensitivity.