• Proceedings of the Korean Vacuum Society Conference
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• 2012.02a
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• pp.558-558
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• 2012

최근의 원자간력현미경(AFM)은 soft한 생체물질을 비파괴적 방법 및 나노크기의 분해능으로 여러 구조적, 물리적 특성 측정이 가능하여 bio분야에 다양이 활용되고 있다. 본 연구에서는 AFM을 이용하여 줄기세포인 BM MSC(bone marrow mesenchymal stem cell)가 신경세포로 분화 여부를 측정하는 방법을 보고하고자 한다. 신경세포의 신호전달은 시냅스에서 신경전달물질을 매개로 하여 이루어지는데, 신경전달물질 중에 D-Glutamic acid는 시냅스후세포에서 흥분성 전위 크기를 증가시킨 상태를 장기간 유지시켜주는 물질로, 특정물질인 Glutamate와 항원-항체 결합을 한다. 본 연구에서는 이 두 물질간의 항원-항체 반응을 활용하여 줄기세포의 신경세포로 분화 여부를 AFM으로 측정하였다. 먼저, 수용성 시료인 두 물질을 증류수에 용해시켜 Mica 기판에 그 용액을 떨어뜨려 자연건조로 시료를 준비한 후, AFM으로 형태 및 크기를 측정하였다. D-Glutamic acid와 Glutamate는 구형 입자 형태를 보였으며, Glutamate의 너비는 ~100 nm이고, D-Glutamic acid는 ~50 nm였다. 두 물질이 든 용액을 섞었을 때, 항원-항체 반응에 의해 다른 크기의 두 구형입자가 붙어 있는 형태가 관찰되었다. 이 반응을 활용하여, 신경세포에서 분비되는 신경전달물질인 D-Glutamic acid를 선별하였다. DMEM 배지에 신경암세포주인 SH-SY5Y 를 접종한 후 $37.6^{\circ}C$의 incubator에서 24시간 배양하고, 화학적 자극(60~70 mM의 KCl 용액을 주입함)을 주어 신경전달물질 분비를 유도하였다. 그 배지에 항체 Glutamate 를 주입하여 자연건조 시킨 후 항원-항체 결합특성을 AFM으로 측정하여, 항원-항체 결합된 이미지와 동일함을 확인하였다. 결과적으로 AFM을 이용한 신경전달물질의 항원-항체 결합여부 측정을 통해, BM MSC 줄기세포의 신경세포로 분화를 판단할 수 있으며, 이 방법은 줄기세포의 특정 세포로의 분화 여부 판단에 활용될 것으로 기대된다.

• Journal of Applied Biological Chemistry
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• v.54 no.1
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• pp.1-6
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• 2011

${\gamma}$-PGA(poly-${\gamma}$-glutamic acid) is an unusual anionic polypeptide that is made of D- and L-glutamic acid units connected by amide linkages between ${\alpha}$-amino and ${\gamma}$-carboxylic acid groups. ${\gamma}$-PGA has been isolated from many kinds of organisms. Many Bacillus strains produce ${\gamma}$-PGA as a capsular material of an extracellular viscous material. It is safe for eating as a viscosity element of fermented soybean products such as Chungkookjang and Natto. It is biodegradable, edible and nontoxic toward humans and the environment and its molecular weight varies from ten thousand to several hundred thousand depending on the kinds of strains used. Therefore, potential applications of ${\gamma}$-PGA and its derivatives have been of interest in the past few years in a broad range of industrial fields such as food, cosmetics, medicine, water-treatment, etc. In this study, a bacterium, Bacillus subtilis GS-2 isolated from the Korean traditional seasoning food, Chungkookjang could produce a large amount of ${\gamma}$-PGA with high productivity and had a simple nutrient requirement. Based on carbon utilization pattern and partial 16S rRNA sequence analysis, the GS-2 strain was identified as B. subtilis. The determination of purified ${\gamma}$-PGA was confirmed with thin layer chromatography (TLC), high performance liquid chromatography (HPLC), fourier transform infrared (FT-IR) spectra, and $^1H$-nuclear magnetic resonance ($^1H$-NMR) spectroscopy.

• Food Science and Preservation
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• v.17 no.6
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• pp.820-825
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• 2010

We investigated changes in the quality characteristics of soy sauce made using salts from deep ocean water; such salts are more economic to obtain than sun-dried salt. pH level and buffer action both decreased as fermentation proceeded. The content of the pure extract after 120 d of fermentation was 7.5%(w/w), and increased as fermentation proceeded. Total and amino acid nitrogen levels after 120 d of fermentation were 1.4% and 0.75%, respectively (both w/w). Of the free amino acids (179.8-315.7 mg%), glutamic acid was the most abundant, followed by alanine, lysine, and leucine. The glutamic acid/total free amino acid ratio was 21.0-23.2%. The content of essential amino acids was 78.0-142.3 mg%; this level increased as fermentation progressed.

• Journal of Microbiology and Biotechnology
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• v.27 no.5
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• pp.1032-1037
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• 2017

The poly-${\gamma}$-$\small{D}$-glutamic acid (PGA) capsule, a major virulence factor of Bacillus anthracis, provides protection of the bacterium from phagocytosis and allows its unimpeded growth in the host. We investigated crosstalk between murine natural killer (NK) cells and macrophages stimulated with the PGA capsule of Bacillus licheniformis, a surrogate of the B. anthracis capsule. PGA induced interferon-gamma production from NK cells cultured with macrophages. This effect was dependent on macrophage-derived IL-12 and cell-cell contact interaction with macrophages through NK cell receptor NKG2D and its ligand RAE-1. The results showed that PGA could enhance NK cell activation by inducing IL-12 production in macrophages and a contact-dependent crosstalk with macrophages.

• Applied Biological Chemistry
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• v.35 no.3
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• pp.210-217
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• 1992

A photosynthetic bacterium strain KUP-74 producing high amount of S-amino-levulinic acid(ALA) was isolated from soils, which was identified as Rhodocyclus gelatinosus. After 10 days cultivation under anaerobic-light condition at $30^{\circ}C$, 4 Klux and pH 6.8, 5 mg/l of ALA was formed extracellularly. ALA productions were increased up to 8 mg/l and 12 mg/l in cell cultivations either by the addition of 0.5% glycerol (v/v) or 10 mM of glycine and succinic acid, respectively, using Lascelles basal medium eliminated L-glutamic acid. By cultivation in the presence of 30 mM each D,L-glutamic acids and D,L-glutamines the yield of ALA showing a late induction phenomenon was reached the maximum value of 21 mg/l. Different culture times were needed to generate maximum ALA yields by the addition of initial precursors of $C_4$ and $C_5$ pathways in basal medium, as being 107 h and 262 h, respectively. 40 mg/l yield of ALA was observed by cell cultivation with the basal medium containing each 10 mM levulinic acid(LA) and glycine simultaneously.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.723-724
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• 2005

• YAKHAK HOEJI
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• v.43 no.3
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• pp.300-305
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• 1999

A sensitive and efficient assay method was applied to determine the level of glutamic acid (GA) and ${\gamma}-aminobutyric$ acid (GABA) in frontal cortex and hippocampus of rat administrated with ethanol and drugs. The compounds were derivatized with ο-phthalaldehyde (OPA) and 2-mercaptoethanof for precolunm analysis. The condition for the simultaneous determination of GA, GABA and $\beta-aminobutyric$ acid (BABA) by high performance liquid chromatography with electrochemical detection was reverse phase $C_{18}$ column as stationary phase, 0.1 M phosphate buffer containing 0.1 mM $Na_4EDTA$ : methanol = 55:45 (v+v) pH 3.8 as mobile phase and 0.7V electrode voltage. The stability of reaction product of GA, GABA and BABA with OPA could be increased by adding the same volume of polyethylene glycol 400 to reaction mixture. The GABA level in frotal cortex of rat was significantly decreased by the administration of picrotoxin and diazepam, but it was significantly increased by the administration of red ginseng total saponin, N-methyl-D-glucamine and (-)-deprenyl.

• Microbiology and Biotechnology Letters
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• v.43 no.3
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• pp.219-226
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• 2015

Acetic acid bacteria (AAB) were isolated from vinegar fermented through traditional methods in Namhae county, Gyeongnam, the Republic of Korea. The isolated strains were Gram negative, non-motile, and short-rods. Three selected strains were identified as either Acetobacter pasteurianus or Acetobacter aceti by 16S rRNA gene sequencing. A. pasteurianus NH2 and A. pasteurianus NH6 utilized ethanol, glycerol, D-fructose, D-glucose, D-mannitol, D-sorbitol, L-glutamic acid and Na-acetate. A. aceti NH12 utilized ethanol, n-propanol, glycerol, D-mannitol and Na-acetate. These strains grew best at 30℃ and an initial pH of 3.4. They were tolerant against acetic acid at up to 3% of initial concentration (v/v). The optimum conditions for acetic acid production were 30℃ and pH 3.4, with an initial ethanol concentration of 5%, resulting in an acetic acid concentration of 7.3−7.7%.

• Food Science and Preservation
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• v.13 no.5
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• pp.661-667
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• 2006

This study was carried out to analyze the major chemical component of the leaf and stem of dried Petasites japonicus S. et Z. Max.. Comparing proximate composition of leaf and stem of Petasites japonicus as dry matter basis, stem contained higher moisture, crude fat, crude ash and carbohydrate, with less crude protein. The main component of free sugar and disaccharide in both leaf and stem were fructose and sucrose, respectively. Total amino acids of leaf and stem were 6,853.32 mg% and 2,350.61 mg% respectively. Although the amino acid composition of leaf and stem were different glutamic acid and aspartic acid were the major amino acids in samples. The major fatty acids of total lipids were linolenic acid in leaf and linoleic acid in stem. The ratios of unsaturated fatty acids to saturated fatty acid were 3.93 in leaf and 3.44 in stem. The unsaturated fatty acid content of samples were 3 times higher than those of saturated fatty acid contents. Oxalic acid was the major organic acids in leaf and stem. The contents of vitamin A, C and E were higher in leaf than in stem. The mined compositions of both leaf and stem were composed in order of K, Mg, Ca, Fe, Na, and Zn.

• Journal of Microbiology and Biotechnology
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• v.31 no.3
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• pp.447-455
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• 2021

Strains of four Bacillus spp. were respectively inoculated into sterilized soybeans and the free amino acid profiles of the resulting cultures were analyzed to discern their metabolic traits. After 30 days of culture, B. licheniformis showed the highest production of serine, threonine, and glutamic acid; B. subtilis exhibited the highest production of alanine, asparagine, glycine, leucine, proline, tryptophan, and lysine. B. velezensis increased the γ-aminobutyric acid (GABA) concentration to >200% of that in the control samples. B. sonorensis produced a somewhat similar amino acid profile with B. licheniformis. Comparative genomic analysis of the four Bacillus strains and the genetic profiles of the produced free amino acids revealed that genes involved in glutamate and arginine metabolism were not common to the four strains. The genes gadA/B (encoding a glutamate decarboxylase), rocE (amino acid permease), and puuD (γ-glutamyl-γ-aminobutyrate hydrolase) determined GABA production, and their presence was species-specific. Taken together, B. licheniformis and B. velezensis were respectively shown to have high potential to increase concentrations of glutamic acid and GABA, while B. subtilis has the ability to increase essential amino acid concentrations in fermented soybean foods.