• Food Science of Animal Resources
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• v.31 no.6
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• pp.952-959
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• 2011

A new bacteriocin-producing lactic acid bacteria (LAB) which has been isolated from kimchi was identified as Pediococcus damnosus by use of API kit and 16S rDNA sequencing, and designated as P. damnosus JNU 534. The bacteriocin produced by P. damnosus JNU 534 markedly inhibited the growth of some of LAB and Listeria monocytogenes, whereas other pathogens including Gram negative bacteria were not susceptible. The production of bacteriocin started at the beginning of exponential phase and reached maximum activity at the early stationary phase. The bacteriocin was stable on the wide pH range of 2-9 and heat treatment up to $100^{\circ}C$ for 15 min. The antimicrobial compound was inactivated by treatments of proteolytic enzymes indicating its proteinaceous in nature. The bacteriocin was purified by 30% ammonium sulfate precipitation followed by hydrophobic interaction column and $C_{18}$ column chromatography. The estimated molecular weight of the bacteriocin using tricine SDS-PAGE was approximately 3.4 kDa and the identified N-terminal amino acid sequence was $NH_2$-ILLEELNV.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.3
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• pp.198-204
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• 2000

In order to utilize by-products which would normally be discarded in marine processing plants, cod teiset protein was hydrolyzed and antioxidative actiTity of the hydrolysate was investigated. AntioxidatiTe peptide was isolated using ultrafiltration membrane, ion-exchange chromatography on a SP-Sephadex C-25 column, gel filtration on a Sephadex G-15 column, high performance liquid chromatography on an ODS column, and capillary electrophoresis chromatography. Antioxidative activities of the cod teiset hydrolysate were compared with ${\alpha}-tocopherol$, one of the commercial antioxidant. The hydrolysate passed through a membrane with molecular weight cut-off (MWCO) 1 kDa was shown the strongest antioxidative activity, and the activity was higher $10{\%}$ as compared with ${\alpha}-tocopherol$. In addition, the peptide isolated by ion-exchange chromatography, gel filtration, and HPLC, respectively, was higher $53{\%}$ as compared with ${\alpha}-tocopherol$, and the amino acid sequence was Ser-Asn-Pro-Glu-Trp-Ser-Trp-Asn.

• The Korean Journal of Mycology
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• v.35 no.1
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• pp.28-32
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• 2007

Fruiting bodies of Inonotus obliquus were collected from the trunk of Betula ermani at 1,100 m of Mt. Odae. Diameter range of the trees at breast height (DBH) was $10{\sim}50$ cm and size range of the sclerotia was $8{\times}5{\sim}20{\times}16cm$. Relationships between the examined strains and Inonotus obliquus strain registered in National Center for Biotechnology Information (NCBI) were very near. And all of 10 strains except strains registered in NCBI showed high homologous characteristics by neighbour joining analysis of ITS sequence. Mycelial growth showed a big difference among strains. Mycelial growth of KFRI 744 was fastest and KFRI 739 was slowest. Difference of mycelial growth between KFRI 735 and 738 was slight, but the difference of mycelial growth between KFRI 744 and 739 was almost twice. Also weight reduction rate among strains showed some difference. KFRI 744 was highest and KFRI 741 was lowest. Vegetative incompatibilities were observed in all mycelial pairings except for KFRI 740-741 and KFRI 742-743 combinations.

• KSBB Journal
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• v.21 no.4
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• pp.279-285
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• 2006

We investigated the effects of immobilization chemistry on the yield of immobilization and the bioactivity of the immobilized enzymes. Trypsin as a model protein and macroporous polymer beads(Toyopearl AF 650M, Tosho Co., Japan) was used as a model matrix. Four methods were used to immobilize trypsin; covalent conjugation by reductive amination(at pH 10.0 and pH 4.0) and affinity interaction via streptavidin-biotin, and double-affinity interaction via biotin-streptavidin-biotin system. The covalent conjugation immobilized $3{\sim}4$ mg/ml-gel, ca. 3-fold higher than the affinity method. However, the specific activity of the covalently(pH 10.0) and affinity-immobilized trypsin(via streptavidin-biotin) are ca. 37% and 50%, respectively, of that of the soluble enzyme(on the low-molecular-weight BAPNA substrate). When the molecular size of a substrate increased, the affinity-immobilized trypsin showed higher clavage activity on insulin and BSA. This result seemed to indicate the streptavidin-biotin system allowed more steric flexibility of the immobilized trypsin in its interaction with a substrate molecule. To confirm this, we studied the molecular flexibility of immobilized trypsin using quartz crystal microbalance-dissipation. Self-assembled monolayers were formed on the Q-sensor surface by aminoalkanethiols, and gultaraldehyde was attached to the SAMs. Trypsin was immobilized in two ways: reductive amination(at pH 10.0) and the streptavidin-biotin system. The dissipation shift of the affinity-immobilized trypsin was $0.8{\times}10^{-6}$, whereas that of the covalently attached enzyme was almost zero. This result confirmed that the streptavidin-biotin system allowed higher molecular flexibility. These results suggested that the bioactivity of the immobilized enzyme be strongly dependent on its molecular flexibility.

• Journal of Pharmaceutical Investigation
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• v.36 no.1
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• pp.59-65
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• 2006

The purpose of this study was to evaluate the bioequivalence of two amlodipine maleate tablets, SKAD tablet (SK Pharma. Co., Ltd., Seoul, Korea, reference drug) and A-PINE tablet (Daewon Pharm. Co., Ltd., Seoul, Korea, test drug), according to the guidelines of Korea Food and Drug Administration (KFDA). Twenty-four healthy male volunteers, $22.79\;{\pm}\;1.86$ years in age and $70.08\;{\pm}\;8.68$ kg in body weight, were divided into two groups and a randomized $2\;{\time}\;2$ crossover study was employed. After a tablet containing 6.42 mg of amlodipine maleate was orally administrated, blood was taken at predetermined time intervals over a period of 144 hr and concentrations of amlodipine in plasma were monitored using LC-MS/MS. Pharmacokinetic parameters such as $AUC_t$ (the area under the plasma concentration-time curve from time zero to 144 hr), $C_{max}$ (maximum plasma drug concentration) and $T_{max}$ (time to reach $C_{max}$) were calculated and analysis of variance (ANOVA) test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t$ and $C_{max}$, and untransformed $T_{max}$. No significant sequence effect was found for all of the bioavailability parameters indicating that the crossover design was properly performed. The 90% confidence intervals of the $AUC_t$ ratio and the $C_{max}$ ratio for A-PINE/SKAD were $log\;0.9429{\sim}log \;1.1476$ and $log\;0.9l46{\sim}log\;1.1488$, respectively. Since these values were within the acceptable bioequivalence intervals of $log\;0.80{\sim}log\;1.25$, recommended by KFDA, it was concluded that A-PINE tablet was bioequivalent to SKAD tablet, in terms of both rate and extent of absorption.

• Journal of Pharmaceutical Investigation
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• v.36 no.1
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• pp.67-73
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• 2006

Lornoxicam is a nonsteroidal anti-inflammatory drug that decreases prostaglandin synthesis by inhibiting cyclooxygenase. It has analgesic, antipyretic and antiinflammatory effects. The purpose of the present study was to evaluate the bioequivalence of two lornoxicam tablets, $Xefo^{\circledR}$ (Hyundai Pharmaceutical Ind. Co., Ltd.) and Lornocam (Samchundang Pharmaceutical Co., Ltd.), according to the guidelines of the Korea Food and Drug Administration (KFDA). The release of lornoxicam from the two lornoxicam formulations in vitro was tested using KP VIII Apparatus II method with various dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty eight healthy male subjects, $24.39{\pm}1.95$ years in age and $68.63{\pm}7.25$ kg in body weight, were divided into two groups and a randomized $2\;{\time}\;2$ cross-over study was employed. After a single tablet containing 4 mg as lornoxicam was orally administered, blood samples were taken at predetermined time intervals and the concentrations of lornoxicam in serum were determined using HPLC with UV detector. The dissolution profiles of two formulations were similar in all tested dissolution media. The pharmacokinetic parameters such as $AUC_t,\;C_{max}\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AUC_t,\;C_{max}$ and untransformed $T_{max}$. The results showed that the differences between two formulations based on the reference drug, $Xefo^{\circledR},$ were -1.56%, 2.16% and -17.12% for $AUC_t,\;C_{max}\;and\;T_{max},$ respectively. There were no sequence effects between two formulations in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of log 0.8 to log 1.25 (e.g., $log\;0.90{\sim}log\;1.05$ and $log\; 0.88{\sim}log\;1.17$ for $AUC_t\;and\;C_{max},$ respectively). Thus, the criteria of the KFDA bioequivalence guideline were satisfied, indicating Lornocam tablet was bioequivalent to $Xefo^{\circledR}$ tablet.

• 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 Microbiology and Biotechnology
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• v.23 no.6
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• pp.856-863
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• 2013

Application of rhizospheric fungi is an effective and environmentally friendly method of improving plant growth and controlling many plant diseases. The current study was aimed to identify phytohormone-producing fungi from soil, to understand their roles in sesame plant growth, and to control Fusarium disease. Three predominant fungi (PNF1, PNF2, and PNF3) isolated from the rhizospheric soil of peanut plants were screened for their growth-promoting efficiency on sesame seedlings. Among these isolates, PNF2 significantly increased the shoot length and fresh weight of seedlings compared with controls. Analysis of the fungal culture filtrate showed a higher concentration of indole acetic acid in PNF2 than in the other isolates. PNF2 was identified as Penicillium sp. on the basis of phylogenetic analysis of ITS sequence similarity. The in vitro biocontrol activity of Penicillium sp. against Fusarium sp. was exhibited by a 49% inhibition of mycelial growth in a dual culture bioassay and by hyphal injuries as observed by scanning electron microscopy. In addition, greenhouse experiments revealed that Fusarium inhibited growth in sesame plants by damaging lipid membranes and reducing protein content. Co-cultivation with Penicillium sp. mitigated Fusarium-induced oxidative stress in sesame plants by limiting membrane lipid peroxidation, and by increasing the protein concentration, levels of antioxidants such as total polyphenols, and peroxidase and polyphenoloxidase activities. Thus, our findings suggest that Penicillium sp. is a potent plant growth-promoting fungus that has the ability to ameliorate damage caused by Fusarium infection in sesame cultivation.

• Parasites, Hosts and Diseases
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• v.52 no.1
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• pp.93-97
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• 2014

Subolesin (4D8), the ortholog of insect akirins, is a highly conserved protective antigen and thus has the potential for development of a broad-spectrum vaccine against ticks and mosquitoes. To date, no protective antigens have been characterized nor tested as candidate vaccines against Dermacentor silvarum bites and transmission of associated pathogens. In this study, we cloned the open reading frame (ORF) of D. silvarum 4D8 cDNA (Ds4D8), which consisted of 498 bp encoding 165 amino acid residues. The results of sequence alignments and phylogenetic analysis demonstrated that D. silvarum 4D8 (Ds4D8) is highly conserved showing more than 81% identity of amino acid sequences with those of other hard ticks. Additionally, Ds4D8 containing restriction sites was ligated into the pET-32(a+) expression vector and the recombinant plasmid was transformed into Escherichia coli rosetta. The recombinant Ds4D8 (rDs4D8) was induced by isopropyl ${\beta}$-D-thiogalactopyranoside (IPTG) and purified using Ni affinity chromatography. The SDS-PAGE results showed that the molecular weight of rDs4D8 was 40 kDa, which was consistent with the expected molecular mass considering 22 kDa histidine-tagged thioredoxin (TRX) protein from the expression vector. Western blot results showed that rabbit anti-D. silvarum serum recognized the expressed rDs4D8, suggesting an immune response against rDs4D8. These results provided the basis for developing a candidate vaccine against D. silvarum ticks and transmission of associated pathogens.

• Korean Journal of Weed Science
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• v.16 no.4
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• pp.337-345
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• 1996

This study was conducted to investigate the protox activity, the PPIX accumulation and the activity of antioxidative enzymes of the oxyfluorfen-tolerant and -susceptible rice cultivars by oxyfluorfen and oxytluorfen-similar herbicides treatment. When treated with acifluorfen, bifenox or oxadiazon, the oxyfluorfen-tolerant rice cultivars showed less decreased in fresh weight than the susceptible rice cultivars. The inhibition of protox activity was in the order of acifluorfen > oxyfluorfen > bifenox > oxadiazon, and the PPIX accumulation was in the sequence of oxadiazon > acifluorfen > oxyfluorfen > bifenox. The inhibition of protox activity and the PPIX accumulation by the herbicide was greater in the susceptible rice cultivars than in the tolerant rice cultivars. The effect inhibiting the decrease of chlorophyll content resulting from the treatment of GC, tetrapyrrole biosynthesis inhibitor, was in the order of oxyfluorfen > acifluorfen > bifenox > oxadiazon, and the tolerant rice cultivar had more than the susceptible rice cultivar. In the treatment of DPE and oxadiazon, the activities of MDAR, POX and GR was higher in the tolerant rice cultivar than in the susceptible rice cultivar, and in the case of POX isozyme the activities of main D and E bands increased in the tolerant rice cultivars at tested herbicides but they didn't in the susceptible rice cultivar.