• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2001.06a
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• pp.149-153
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• 2001

To investigate the mode of bactericidal action for antimicrobial peptide, pediocin, synthetic and mutant pediocins were prepared by direct chemical synthesis. Native pediocin was purified from Pedio-coccus acidilactici M and its conformational structure and bactericidal functions were analyzed and compared to synthetic pediocin. Schematic mode of pediocin actions, how pediocin binds on the target cell membrane, penetrates and makes tunnel are proposed. For these purposes, primary and secondary structures of pediocin was analyzed and disulfide bond assignment was also done. The pediocin purified from P. acidilactici M had high effective bactericidal ability against gram positive bacteria, especially Listeria monocytogenes and was very stable at extreme pHs and even at high temperatures such as autoclaving temperature (121$^{\circ}C$). Pediocin was consisted of 44 amino acids with four cysteines. Novel synthetic peptides were achieved by solid phase peptide synthesis(SPPS) method. To explain the function of cysteine in C-terminal region, mutant pediocin, Ped[C24A＋C44A], was synthesized and their structural and biological functions were analyzed. Second mutant pediocin, Ped[KllE], was prepared to explain the function of lysine at 11 of N-terminal part of pediocin, especially loop of $\beta$-sheet, and to predict the initial binding site of pediocin. The native and synthetic pediocins was showed random coil conformation by spectropolarimetry in moderate conditions. This conformation was observed in extreme conditions such as high temperature and low and high pHs, also. Circular dichroism(CD) data also showed the existence of $\beta$-turn structure in N-terminal part both native and synthetic pediocins. A structural model for pediocin predicts that 18 amino acids in the N-terminal part of the peptide assume a three-strand $\beta$-sheet conformation. This random coil in C-terminal part of pediocin was converted to folding structure, helix structure, in nonpolar solvents such as alcohol and TFE. The disulfide bond between $^{9}$ Cys and $^{14}$ Cys was concrete and inevitable, however, evidences of disulfide bond between $^{24}$ Cys and $^{44}$ Cys was not. Data of Ped[C24A＋C44A], pediocin mutant showed that $^{44}$ Cys was required during killing the target cells but not inevitable, since Ped[C24A＋C44A] still have bactericidal activity but much less than native pediocin. Another pediocin mutant, Ped[KllE], had still bactericidal activity, was controversial to propose that positive charge like as $^{11}$ Lys in loop or hinge in bacteriocin bound or helped to binding to microorganism with electrostatic interaction between cell membrane especially teichoic acid and positive amino acid nonspecifically. The conformation of pediocin among native, synthetic and mutant pediocins did not show big difference. The conformations between oxidized and reduced pediocin were almost similar regardless of native or synthetic.

• Korean Journal of Medicinal Crop Science
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• v.24 no.5
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• pp.386-392
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• 2016

Background: Sorghum is a major cereal food crop used in many parts of the world. It has been grown on a subsistence level by farmers, under various conditions of environmental stresses in the semi-arid tropics of Africa and Asia. This plant has received significant attention because of its ability to reduce cholesterol in the blood, and its anti-dementia, antioxidant, and antimicrobial properties. It is possible to develop a functional and commercially viable sorghum variety by using superior cultivars of sorghum. The objective of this study was to build a database of superior sorghum accession. Methods and Results: We used 250 sorghum accessions collected from different geographical bioregions in Korea. We determined various agronomic characters including germination rate and ear length of these accessions. To determine the antioxidant capacity, we measured the 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)-diammonium salt radical scavenging activity, total phenolic contents, and total flavonoid contents. Accession 189 showed higher germination (> 80%) than the other accessions. Higher 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity was observed in 11-SB-078 ($RC_{50}$; $1.89{\pm}2.88{\mu}g/m{\ell}$), and higher 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)-diammonium salt radical scavenging activity was recorded in 11-SB-116 ($RC_{50}$; $35.48{\pm}2.42{\mu}g/m{\ell}$) than in the other accessions. The ear length ranged from 15 cm to 48 cm, the total phenolic contents ranged from $3mg{\cdot}GAE/g$ to $77mg{\cdot}GAE/g$, and total flavonoid contents ranged from $0.09mg{\cdot}QE/g$ to $1.07mg{\cdot}QE/g$. Conclusions: Among 250 sorghum accessions, we selected 10 with both superior agronomic characters and highly functional food quality.

• Korean Journal of Microbiology
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• v.51 no.1
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• pp.48-60
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• 2015

612 strains isolated from Korean traditional fermented food in Sunchang and their investigated biochemical characterization and ability of biogenic amines non-producing. We selected the SCJ4 having various activity by measurement of extracellular enzyme, antioxidant and antimicrobial activities. Selected strain SCJ4 by 16S rRNA sequencing and biochemical characterization was named Bacillus subtilis SCJ4. And then, we investigated cell growth of SCJ4, and optimized of culture medium constituents using response surface methodology as statistically method. Response surface methodology used Plackett-Burman experimental design for screening of medium constituent. Tryptone, peptone and $MgSO_4$ as medium constituent improving cell growth selected. In order to find out optimal concentration on each constituent, we carried out central composite design. Consequently, optimized concentrations of tryptone, peptone and $MgSO_4$ were predicted to be 15.35 g/L, 12.235 g/L, and 3.5 g/L respectively. Through the model verification, we confirmed about 1.28-fold improvement of the dried cell weight from 0.8767 g/L to 1.1222 g/L when compared to basal medium.

• Journal of Life Science
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• v.24 no.8
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• pp.880-886
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• 2014

In this study, we isolate and identify bacteria from seawater collected from Jeju coast, to evaluate the antimicrobial activity against the fish pathogenic bacteria. 14 bacterial strains were isolated and identified using physiological, biochemical and molecular tools. Antibacterial activity of all the 14 isolates were screened against four major fish pathogens namely, two Gram-positive: Streptococcus iniae, Streptococcus parauberis and two Gram-negative: Vibrio anguillarum, Edwardsiella tarda. Results revealed that among the 14 isolates, MK-11 was found to have antibacterial activity against S. iniae, S. parauberis, V. anguillarum Particularly, S. iniae was susceptibility with the MIC value of $250{\mu}g/ml$. The biochemical and physio-chemical results reveal that MK-11 had the sugar-alcohol disassemble ability of the D-sorbitol and D-mannitol. Also the utilization of the yeast extract, sorbitol and di-potassium phosphate were noted to be high. The optimum culture condition such as pH and temperature was recorded as pH 6.0, $25^{\circ}C$ and along with 1% NaCl which differs from the previous reports particularly in nutrient resolutions. As results of the analysis of 16S rDNA sequences, MK-11 show the high similarity with Paenibacillus polymyxa, P. jamilae, P. brasilensis 99.78%, 99.43%, 99.39%, repectively. Hence, in the present study, the isolated Paemibacillus sp. MK-11 from Jeju seawater possesses the antibacterial activity against fish pathogens and it could be used as a new antibiotic agents against the gram positive fish pathogens.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1249-1256
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• 2017

In our search for new sources of bioactive secondary metabolites from Streptomyces sp., the ethyl acetate extracts from endophytic Streptomyces SUK 25 afforded five active diketopiperazine (DKP) compounds. The aim of this study was to characterize the bioactive compounds isolated from endophytic Streptomyces SUK 25 and evaluate their bioactivity against multiple drug resistance (MDR) bacteria such as Enterococcus raffinosus, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumanii, Pseudomonas aeruginosa, and Enterobacter spp., and their cytotoxic activities against the human hepatoma (HepaRG) cell line. The production of secondary metabolites by this strain was optimized through Thornton's medium. Isolation, purification, and identification of the bioactive compounds were carried out using high-performance liquid chromatography, high-resolution mass liquid chromatography-mass spectrometry, Fourier transform infrared spectroscopy, and nuclear magnetic resonance, and cryopreserved HepaRG cells were selected to test the cytotoxicity. The results showed that endophytic Streptomyces SUK 25 produces four active DKP compounds and an acetamide derivative, which were elucidated as $cyclo-({\text\tiny{L}}-Val-{\text\tiny{L}}-Pro)$, $cyclo-({\text\tiny{L}}-Leu-{\text\tiny{L}}-Pro)$, $cyclo-({\text\tiny{L}}-Phe-{\text\tiny{L}}-Pro)$, $cyclo-({\text\tiny{L}}-Val-{\text\tiny{L}}-Phe)$, and N-(7-hydroxy-6-methyl-octyl)-acetamide. These active compounds exhibited activity against methicillin-resistant S. aureus ATCC 43300 and Enterococcus raffinosus, with low toxicity against human hepatoma HepaRG cells. Endophytic Streptomyces SUK 25 has the ability to produce DKP derivatives biologically active against some MDR bacteria with relatively low toxicity against HepaRG cells line.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.2
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• pp.141-147
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• 2008

This study investigated the isoflavone contents and physiological properties of non-fermented soybean (NF) and the fermented soybeans prepared with Asp. oryzae (AO) and B. natto (BN). The total isoflavone contents (daidzin, genistin, daidzein and genistein) of NF, AO and BN were 81.8 mg/100 g, 130.7 mg/100 g and 139.5 mg/100 g, respectively. Especially, the total phenol contents of NF, AO and BN were 2.1%, 4.3% and 7.6%, and the total flavonoid contents were 1.3%, 1.6% and 2.7%, respectively. The nitrite-scavenging abilities of NF, AO and BN were 34.4%, 55.2% and 92.5%, respectively. Antimicrobial activity of BN was shown to be the strongest to Bacillus cereus, Staphylococcus aureus and Escherichia coli. The SOD-like activity was the strongest in AO, whereas the electron donating ability was the strongest in BN. Antioxidant activity of AO at concentration of 0.02% was stronger than BN or NF.

• Korean Journal of Food Science and Technology
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• v.32 no.1
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• pp.168-173
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• 2000

Anticariogenic activity of the unripe apple extract was studied by observing the inhibitory effects on GTase(glucosyltransferase) activity, cell adherence and acid production of Streptococcus mutans. Among the four S. mutans strains, S. mutans MT 8148 had the highest water-insoluble glucan forming activity. (+)-Catechin and tannic acid, the major components of the unripe apple polyphenols inhibited GTase activity by 60% at 1 mg/ml and 90% at 5 mg/ml. Tannic acid and unripe apple extract inhibited adherence ability of S. mutans by 50% and 30%, respectively. But the acid production of S. mutans was not influenced by the polyphenols. Disc diffusion test showed that the polyphenols have no antimicrobial activity against S. mutans, which indicates that the inhibition of GTase activity and cell adherence were not resulted from the cell growth inhibition. Our results convinced the possible application of the unripe apple extract as the anticariogenic food additives.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.6
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• pp.797-804
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• 2005

Physiological activity of citrus essential oil (CEO) was determined to examine possible use of the food processing by-product as a functional material for food and cosmetic composition. The effect of gamma irradiation on the change of physiological activity also investigated at 0, 10 and 20 kGy. Limonene contents of CEO was $88.3\pm1.30\%$. Electron donating ability of CEO was $69\%$. Lipid oxidation was retarded by CEO. CEO showed antimicrobial activity against 1 yeast,4 molds and 4 bacteria species tested. More than $80\%$ of inhibition of cancer cell growth was presented by CEO using A549, HT29, HepG2, B16F10 and G361 cells at a 500 ppm level. Irradiation of CEO did not affect any physiological functions. A Salmonella mutagenicity assay indicated that the 20 kGy irradiated CEO did not show any mutagenicity Therefore, CEO, which is a major by-product in citrus processing, could be used as a functional material in various application.

• Journal of Microbiology and Biotechnology
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• v.26 no.6
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• pp.1140-1147
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• 2016

The plasma and serum of Crocodylus siamensis have previously been reported to exhibit potent antimicrobial, antioxidant, and anti-inflammatory activities. During wound healing, these biological properties play a crucial role for supporting the formation of new tissue around the injured skin in the recovery process. Thus, this study aimed to evaluate the wound healing properties of C. siamensis plasma and serum. The collected data demonstrate that crocodile plasma and serum were able to activate in vitro proliferation and migration of HaCaT, a human keratinocyte cell line, which represents an essential phase in the wound healing process. With respect to investigating cell migration, a scratch wound experiment was performed which revealed the ability of plasma and serum to decrease the gap of wounds in a dose-dependent manner. Consistent with the in vitro results, remarkably enhanced wound repair was also observed in a mouse excisional skin wound model after treatment with plasma or serum. The effects of C. siamensis plasma and serum on wound healing were further elucidated by treating wound infections by Staphylococcus aureus ATCC 25923 on mice skin coupled with a histological method. The results indicate that crocodile plasma and serum promote the prevention of wound infection and boost the re-epithelialization necessary for the formation of new skin. Therefore, this work represents the first study to demonstrate the efficiency of C. siamensis plasma and serum with respect to their wound healing properties and strongly supports the utilization of C. siamensis plasma and serum as therapeutic products for injured skin treatment.

• Food Science of Animal Resources
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• v.30 no.2
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• pp.328-335
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• 2010

The aim of our study was to develop a new starter culture for fermented milk. Polymerase chain reaction screening of 103 acid-producing isolates from Kimchi identified 72 Lactobacillus strains. The ability of the strains to inhibit the growth of the food-borne human pathogens (Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus) was measured, using a conventional paper disk method. Among the 72 strains, strain LHC52 displayed potent antagonistic activity. Use of 16S rDNA sequencing and the API 50CHL system identified the strain as Lactobacillus plantarum and it was designated L. plantarum LHC52. Biochemical analyses revealed especially high antibacterial activity against E. coli. Yogurt produced using L. plantarum LHC52 did not show different microbiological and physicochemical properties compared to conventionally-prepared yogurt, implicating L. plantarum LHC52 as a useful, potently antibacterial starter culture for yogurt preparation.