• Korean Journal of Fisheries and Aquatic Sciences
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• v.32 no.6
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• pp.779-783
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• 1999

The process of alginate preparation was shortened with microwave pretreatment and hot water extraction without any other treatment. The yield of extracted alginate was very different according to raw material and microwave treatment conditions. The yield of extracted alginate from sea mustard was higher than sea tangle and increased with the temperature rise and time of microwave treatment. Optimum extraction conditions by microwave treatment were 90 min at $100^{\circ}C$ for the sea mustard and 120 min at $100^{\circ}C$ for the sea tangle. On the other hand, the solubility of the alginate from sea mustard was $69\%\~80\%$ and the viscosity were 8.7$\~$1.5 cps, respectively. The solubility and the viscosity of the sea tangle alginate was higher than the sea mustard to $77\~84\%$ and in the range of 8.9$\~$1.8 cps, respectively. The extraction temperature by hot water greatly influenced on the yield of alginate, but the solubility and viscosity was not affected by the hot water extraction. The molecular weight of alginate obtained through the extraction by $Na_2CO_3$ solution and the microwave pretreatment was 800 kDa, and in the range of 12$\~$45 kDa, respectively.

• Journal of Plant Biotechnology
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• v.30 no.4
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• pp.323-328
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• 2003

Soil salinity is one of the most serious abiotic stresses limiting the productivity of agricultural crops. To cope with salt stress, plants respond with physiological, developmental and biochemical changes, including the synthesis of a number of proteins and the induction of gene expression. Salicornia herbacea is a halophytic plant that grows in salt marshes and on muddy seashores. In order to understand the biochemical and molecular mechanisms of salt tolerance in S. herbacea, we isolated several genes that involved in the salt tolerance by mRNA differential display. Here we report the cloning of a cDNA encoding fructose-1, 6-bisphosphate aldolase, named ShADL, which is 1293 bp long and contains an open reading frame consisted of 359 amino acids with calculated molecular mass of 39 kDa. ShADL protein showed 86％ identity with Arabidopsis and 78％ with aldolase of common ice plant. Northern blot analysis revealed that the transcript of ShADL gene was increased dramatically depending on the NaCl concentrations.

• The Korean Journal of Mycology
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• v.27 no.4 s.91
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• pp.293-298
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• 1999

An one percent sodium carbonate extract prepared from sclerotia of Poria cocos activated the proliferation of the T lymphocytes as measured by mixed lymphocyte responses(MLR). The active fraction, PCSC22, was isolated from an one percent sodium carbonate extract by a combination of fractionation procedures, including ethanol precipitation and chromatographies on column of DEAE-cellulose and Sephadex G50. Carbohydrate and peptide contained in PCSC22 were 78 : 22% in ratio. On employing gel filtration high performance liquid chromatography, PCSC22 exhitited a homogeneous peak with an average molecular weight of 8 kDa. The sugar moiety of PCSC22 was composed with mannose (92%), galactose (6.2%) and arabinose (1.3%), which might be indicated as heteromannan. Fifteen amino acids were found in peptide moiety of the polysaccharide and aspartic acid, serine, and valine were major components. PCSC22 activated the primary proliferation of T lymphocytes measured by mixed lymphocyte responses, the antibody production of the B lymphocytes and the secretion of nitric oxide from macrophage cell line, RAW264.7.

• Journal of Life Science
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• v.25 no.2
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• pp.180-188
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• 2015

The bacterial strain isolated from Kimchi showed antibacterial activity against Micrococcus luteus IAM 1056. The selected strain was identified as Lactococcus lactis by 16S rRNA nucleotide sequence analysis and named as Lactococcus sp. KD 28. The treatment of culture supernatant with proteinase K removed antibacterial activity, indicating its proteinaceous nature, a bacteriocin. This bacteriocin was sensitive to hydrolytic enzymes such as ${\alpha}$-chymotrypsion, trypsin, proteinase K, lipase, ${\alpha}$-amylase and subtilisin A. The bacteriocin was highly thermostable and resistant to heating at $80^{\circ}C$ for up to an hour but 50 % of the total activity was remained at $100^{\circ}C$ for 30 min. The pH range from 2.0 to 8.0 had no effect on bacteriocin activity and it was not affected by solvents such as acetonitrile, isopropanol, methanol, chloroform and acetone up to 50% concentration. The bacteriocin showed antibacterial activity against M. luteus IAM 1056, Lactobacillus delbrueckii subsp. lactis KCTC 1058, Enterococcus faecium KCTC 3095, Bacillus cereus KCTC 1013, B. subtilis KCTC 1023, Listeria ivanovii subsp. ivanovii KCTC 3444, Staphylococcus aureus subsp. aureus KCTC 1916, B. megaterium KCTC 1098 and B. sphaericus KCTC 1184. The bacteriocin was purified through ammonium sulfate concentration, SP-Sepharose chromatography and RP-HPLC. The molecular weight was estimated to be about 3.4 kDa by tricine-SDS-PAGE analysis.

• Korean Journal of Veterinary Research
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• v.45 no.1
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• pp.45-53
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• 2005

GroE that is a heat shock protein composed of GroEL and GroES is known as an immunodominant target of both the humoral and cellular immune responses in bovine brucellosis. This study was carried out to characterize groE gene encoding heat shock proteins of B. abortus isolated in Korea and to evaluate the immunogenicity of the GroE protein expressed in E. coli system. In PCR the specific signals with the size of 2,077 bp were detected in five strains isolated from the mammary lymphnodes of the dairy cattle that were serologically positive and the reference strains. In comparison of the sequences of nucleotides and amino acids among the strains, GroES showed 100% identity in both sequences. GroEL was evaluated 99.0~99.9% in nucleotides and 98.0~100% homology in amino acids. The groE gene including groES and groEL was inserted into pET29a vector and constructed pET29a-GroE recombinant plasmids. The inserted groE was confirmed by digestion with Nco1 and EcoR1 endonucleases and nucleotide sequencing. E. coli BL (DE3) was transformed with pET29a-GroE, named as E. coli BL (DE3)/pET29a-GroE. In SDS-PAGE, it was evident that the recombinant plasmid effectively expressed the polypeptides for GroES (10 kDa) and GroEL (60 kDa) in 0.5, 1 and 2 hours after IPTG induction. The immuno-reactivity of the expressed proteins were proved in mouse inoculation and Western blot analysis.

• Tuberculosis and Respiratory Diseases
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• v.77 no.2
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• pp.73-80
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• 2014

Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

• Food Science and Preservation
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• v.24 no.6
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• pp.820-826
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• 2017

Soybean is one of the most common food materials for making traditional Korean foods such as soybean paste, soy source and soy snack, and their manufacturing processes include heat treatment of soybean. This study was carried out to investigate the effect of heat treatment on the physicochemical properties of soybean. All samples were heat treated under commercial steamed, puffed or air-fried conditions, and then the protein molecular weight distribution, thermal properties, fluorescence intensity, protein solubility, and water and oil holding ability of the heat treated soybeans were examined. Sodium dodecyl sulfate polyacrylamide gel electrophoresis indicated that heat treatment caused fragmentation of polypeptide chain in soybean, showing the band of low molecular ranging from 17 to 40 kDa. The differential scanning calorimetric analysis showed the decrease of enthalpy values (${\Delta}H$) by heat treatment. Fluorescence spectroscopy indicated that the heat treatment caused lipid oxidation as proved by increasing emission intensity. The protein solubility at pH 3-6, and water holding capacity of heat treated soybeans were the higher than no treatment. These results suggest that the heat treatment resulted in decreased enthalpy values, and increased protein degradation, lipid oxidation and water affinity of soybean. Moreover, the effect of heat treatment on physiochemical properties of soybeans was more significant under air-fried condition.

• Analytical Science and Technology
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• v.17 no.2
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• pp.163-172
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• 2004

The molecular size distribution and fluorescence properties of soil fulvic acids (FA) were characterized by using gel filtration chromatography (GFC) and luminescence spectroscopy. The objectives of this work were to fractionate the FA extracted from a forest soil into different nominal molecular size using GFC system and to characterize the fluorescence properties (excitation, emission and synchronous) of these fractions using luminescence spectrometer. The GFC column was calibrated with polyethylene glycols, acetone and dextrane Blue. The total permeation volume of the GFC system was 404 mL and the void volume 130 mL. The GFC molecular weight of the soil FA was in the range of 190~8,900 Dalton and the molecular weight at the peak on the chromatogram was 930 Dalton. The fluorescence intensity ratio ($I_{498nm}/I_{390nm}$) was found to be increased with an increasing molecular weight. This results may suggest that the fulvic acid fractions with high molecular weight have large amount of the condensed aromatic compound.

• Molecules and Cells
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• v.37 no.3
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• pp.226-233
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• 2014

Excessive reactive oxygen species (ROS) generated from abnormal cellular process lead to various human diseases such as inflammation, ischemia, and Parkinson's disease (PD). Sensitive to apoptosis gene (SAG), a RING-FINGER protein, has anti-apoptotic activity and anti-oxidant activity. In this study, we investigate whether Tat-SAG, fused with a Tat domain, could protect SH-SY5Y neuroblastoma cells against 1-methyl-4-phenylpyridinium ($MPP^+$) and dopaminergic (DA) neurons in the substantia nigra (SN) against 1-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine (MPTP) toxicity. Western blot and immunohistochemical analysis showed that, unlike SAG, Tat-SAG transduced efficiently into SH-SY5Y cells and into the brain, respectively. Tat-SAG remarkably suppressed ROS generation, DNA damage, and the progression of apoptosis, caused by $MPP^+$ in SH-SY5Y cells. Also, immunohistochemical data using a tyrosine hydroxylase antibody and cresyl violet staining demonstrated that Tat-SAG obviously protected DA neurons in the SN against MPTP toxicity in a PD mouse model. Tat-SAG-treated mice showed significant enhanced motor activities, compared to SAG- or Tat-treated mice. Therefore, our results suggest that Tat-SAG has potential as a therapeutic agent against ROS-related diseases such as PD.

• Journal of Life Science
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• v.12 no.1
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• pp.96-105
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• 2002

To investigate expression of the artificial gene encoding a repeated tripeptide lysyl-glutamyl-tryptophan in tobacco plant, the plant binary vector, pART404 has been constructed, which contains the duplicated CaMV 35S promoter, an artificial gene coding for repetitive polymer (Lys-Glu-Trp)$_{64}$, and nopaline synthase (nos) terminator. The recombinant expression vector was introduced in Nicotiana tabacum (var. Xanthi) via Agrobacterium tumefaciens-mediated trans-formation. The transgenic calli selected by kanamycin containing medium were then regenerated to whole plants. Southern blot analysis indicated that five transgenic plants (No. 1, 7, 9, 43, 45) showed the hybridizing signals at 1.1 kb of the expected size on EcoRI digestion and each of the transgenic plants contained 1 or 3 copies of the artificial gene inserted into its genome. By northern blot analysis, the size of the hybridized total RNA was estimated to be approximately 1.2 kb and the RNA appeared generally to have the integrity. Western blot indicated that the protein was detected at the position of 33 kDa and the expression level of the polypeptide in the transgenic plant (No. 45) was measured to approximately 0.1% of the total protein.