• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.45-46
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• 2000

After cell removal from fermentation broth the supernatant containing poly (${\gamma}-glutamic acid$) (${\gamma}-PGA$) was concentrated by ultrafiltration in order to reduce the amount of organic solvents such as methanol, ethanol, propanol required for the recover of ${\gamma}-PGA$ with precipitation. The concentration and volume reduction of cell-free solution by ultrafiltration could reduce 3 times the amount of ethanol for the recover of ${\gamma}-PGA$ from cell-free fermentation broth.

• 한국식품영양과학회지
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• 제35권3호
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• pp.255-261
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• 2006

칼슘은 한국인에게 가장 부족되기 쉬운 영양소로 섭취만큼이나 흡수가 중요한 요인이 되는 미네랄이다. 최근 칼슘흡수 증진에 효과가 있는 소재로 주목받고 있는 폴리 감마글루탐산(${\gamma}-PGA;\;poly-{\gamma}-glutamic\;acid)$이 체내 칼슘흡수에 미치는 영향을 알아보기 위해 4주령된 수컷 흰쥐(Sprague Dawley, female rats)를 이용하여 3주간 칼슘결핍식이를 급여시켜 칼슘결핍을 유도 후에, ${\gamma}-PGA$ 보충식이를 4주간 급여하였다. 실험군은 칼슘결핍군(Ca free group), 칼슘보충군(칼슘1.2% 보충군, total Ca 0.45%), 칼슘과 CPPl% 보충군(칼슘1.2%+CPP1% 보충군), 칼슘과 PGA1% 보충군(칼슘1.2%+${\gamma}-PGA1%$ 보충군), 칼슘과 CP1%+PGA1% 보충군(칼슘1.2%+CPP1%+${\gamma}-PGA1%$ 보충군), 칼슘과 PGA3% 보충군(칼슘1.2%+$\gamma$-PGA3% 보충군)으로 구분하였다. 이때 ${\gamma}-PGA$의 효과를 검증하기 위해 ${\gamma}-PGA$와 칼슘흡수 기전이 유사하고 이미 효능이 알려져 있는 casein phosphopeptide 보충군을 실험군에 추가하였다. 식이섭취량은 칼슘결핍군이 다소 낮았으나 유의적인 차이는 없었으며 칼슘섭취량도 유의적 차이 없이 비슷한 수준을 보였다. 변 및 뇨중 칼슘 배설량은 칼슘만을 섭취한 대조군에 비해 ${\gamma}-PGA$보충군에서 유의적으로 낮았다(p<0.05). 칼슘흡수율 또한 그룹간의 유의적 차이를 보여 칼슘만을 섭취한 그룹에 비해 ${\gamma}-PGA$를 함께 보충한 그룹의 흡수율이 30%더 높은 경향을 보였으며 이는 이미 효능이 알려진 CPP보다 우수한 수준이었다. 골형성 지표인 혈중 ALP를 측정한 결과 대조군에 비해 ${\gamma}-PGA$보충군이 유의적으로(p<0.05) 낮았고, 혈중 칼슘농도는 항상성을 유지하며 그룹간의 유의적 차이 없이 비슷한 수준을 보였으며 , OHPr/Cre도 유의적 차이는 없었다. 뼈 중 칼슘함량 및 강도는 칼슘결핍군이 나머지군에 비해 유의하게(p<0.03) 낮았으며, 칼슘함량의 경우 칼슘만을 보충한 그룹에 비해 ${\gamma}-PGA$를 함께 보충한 그룹이 농도 의존적으로 높은 칼슘함량을 보였고(p<0.05), 골강도의 경우는 유의적 차이는 없었으나 ${\gamma}-PGA$를 함께 보충한 그룹이 농도 의존적으로 골강도가 높은 경향을 나타내었다. 결론적으로 칼슘결핍을 유발한 흰쥐 에 칼슘만을 보충시켰을 때보다 칼슘과 함께 ${\gamma}-PGA$를 함께 보충했을 경우, 칼슘흡수 증진 및 골대사에 긍정적으로 기여함을 확인할 수 있었다. 그러나 좀 더 명확한 효과 검증을 위해서는 차후 많은 개체수와 장기간의 식이를 통한 반복실험, ${\gamma}-PGA$섭취수준에 따른 추가실험이 진행되어야 할 것으로 생각된다.

• Preventive Nutrition and Food Science
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• 제13권4호
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• pp.354-361
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• 2008

A novel bacterium isolated from Cheonggukjang was identified as a glutamate-dependent Bacillus subtilis HA with 98.3% similarity to Bacillus subtilis Z99104. Optimization of poly-$\gamma$-glutamic acid ($\gamma$-PGA) production by modulating fermentation factors including carbon sources, nitrogen sources, inorganic salts and fermentation time was investigated. Optimum culture broth for $\gamma$-PGA production consisted of 3% glutamate, 3% glucose and various salts, resulting in the PGA production of 22.5 g/L by shaking culture for 72 hr at $37^{\circ}C$. Average molecular weight of $\gamma$-PGA was determined to be 1,220 kDa through MALLS analysis. The $\gamma$-PGA solution showed a typical pseudoplastic flow behavior, and a great decrease in consistency below pH 6.0 regardless of the same molecular weight of $\gamma$-PGA. The molecular weights of isolated $\gamma$-PGA were drastically decreased by heat treatment in various acidic conditions, resulting in different hydrolysis of $\gamma$-PGA. The consistency of $\gamma$-PGA solution was greatly decreased with increase heating time in acidic conditions.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.919-923
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• 2005

A bacterium capable of poly(${\gamma}$-glutamic acid) production was isolated from nonpasteurized soy sauce. It was judged to be a variety of Bacillus subtilis and designated as B. subtilis C1. B. subtilis C1 produced ${\gamma}$-PGA in the absence of exogenous glutamic acid; therefore, it is a de novo PGA­producing bacterium. The product produced by B. subtilis C1 was characterized by amino acid analysis to be composed of solely glutamic acid. However, the $H^1-NMR$ spectra showed chemical shifts of glycerol protons in addition to those of authentic ${\gamma}$-PGA, indicating that the product is in fact a bioconjugate of ${\gamma}$-PGA. The finding is unique, because the microbial production of ${\gamma}$-PGA bioconjugate has never been reported before. The molecular mass of the product was over 10,000 kDa as determined by GPC, and $97\%$ of the product was D-glutamate, indicating that L-glutamate was converted to its D-form counterpart by B. subtilis C1.

• Journal of Applied Biological Chemistry
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• 제48권4호
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• pp.213-217
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• 2005

Microbial hydrogel was prepared with poly(${\gamma}$-glutamic acid) produced from Bacillus subtilis BS62 using crosslinking reagent, ethylene glycol diglycidyl ether (EGDE), and its physico-chemical characteristics were examined. Hydrogel which prepared from 10 grams of 10% PGA solution with $600\;{\mu}l$ of EGDE at $50^{\circ}C$ for 17 h swelled 4,320 times its dry weight, and time to reach swelling equilibrium in deionized water at 4 to $45^{\circ}C$ range was about 20 h. Swollen hydrogel shrunk in ionic solutions, and rate of shrinkage was higher in calcium chloride solution than sodium chloride solution. Swelling rate of hydrogel increased 1.3-fold of initial swelling rate for 30 min at $80^{\circ}C$.

• Journal of Microbiology and Biotechnology
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• 제20권4호
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• pp.803-808
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• 2010

Poly-${\gamma}$-glutamic acid (${\gamma}$-PGA) is a natural edible polypeptide in which glutamate is polymerized via ${\gamma}$-amide linkages. First, we assessed the eye irritancy potential of ${\gamma}$-PGA in rabbits. Additionally, we studied the effects of ${\gamma}$-PGA on corneal wound healing, due to the anti-inflammatory properties and water retaining abilities of ${\gamma}$-PGA. In this study, the effects of ${\gamma}$-PGA on corneal wound healing after an alkali burn were evaluated. Thirty eyes wounded by alkali burning in 30 white rabbits were divided into three groups: group A was treated with 0.1% 5,000 kDa ${\gamma}$-PGA for 2 days; group B was treated with 0.1% hyaluronic acid; and group C was not treated, as a control. The area of corneal epithelial defect was examined at 12, 24, 30, 36, 42, and 48 h after corneal alkali wounding to determine initial wound healing. We found that ${\gamma}$-PGA promoted corneal wound healing, compared with controls, and showed similar effects to hyaluronic acid. These results indicate that ${\gamma}$-PGA stimulates corneal wound healing by an anti-inflammatory effect and enhancing cell migration and cell proliferation. ${\gamma}$-PGA is a promising biomaterial that may be a substitute for hyaluronic acid in corneal wound healing treatment.

• Macromolecular Research
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• 제13권4호
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• pp.339-343
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• 2005

Microbial hydrogel was prepared by ${\gamma}-irradiation$ of poly(${\gamma}-glutamic acid$) (PGA) which was produced from Bacillus subtilis BS 62 and it's physico-chemical characteristic was examined. The hydrogel, prepared from 10% PGA with the dose of 48 kGy, was swollen up to 1,370 times of specific water content as dry weight basis. The hydrogels obtained above the dose of 48 kGy appeared to have higher compressive strength but lower specific water content. The period to reach a swelling equilibrium for the hydrogel in deionized water at the temperature range of 4 to $45^{\circ}C$ was about 10 h. The swollen hydrogel was shrunk in ionic solutions with the increase of ionic strength, and the rate of shrinkage was greater in calcium chloride solution than in sodium chloride. Specific water content of the hydrogel was quickly decreased at $80^{\circ}C$, showing a thennally hydrodegradable property.

• 한국식품영양과학회지
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• 제46권9호
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• pp.1114-1121
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• 2017

전통장류로부터 식품 유해요소를 생성하지 않는 Bacillus 균주를 분리하여 세포외효소 활성(amylase, protease, cellulase, xylanase)을 측정한 후, 단백질 분해 활성이 우수한 14개 균주와 비교균주 1균주를 선발하였다. 선발된 균주에 대해 16S rRNA 유전자를 이용한 균주 동정을 실시한 결과, B. amyloliquefaciens 10종, B. methylotrophicus 1종, B. velezensis 1종, B. subtilis 3종이 분리 동정되었다. 그중 B. subtilis JBG17019, B. amyloliquefaciens JBD17076, B. amyloliquefaciens JBD17109 균주에서 식중독미생물에 대한 증식 억제능이 확인되었다. Glutamic acid 대사와 관련한 발효특성을 확인하기 위하여 선발된 Bacillus 균주에 대해 glutamate, glutamine 및 ${\gamma}$-PGA 생성능을 측정하였다. 발효특성과 ${\gamma}$-PGA 생성능에 대한 다변량 요인분석을 주성분(PCA) 추출법으로 분석한 결과, PC1(효소 활성(amylase, cellulase, xylanase), PC2(${\gamma}$-PGA 생성능) 및 PC3(protease, glutamate 및 glutamine)의 3가지 주성분이 분류되었다. 주성분(PC)의 추출에 따라 B. amyloliquefaciens JBD17076 및 B. subtilis JBG17019 균주는 우수한 효소 활성 및 ${\gamma}$-PGA 생성을 하는 것으로 평가되었다.

• Journal of Microbiology and Biotechnology
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• 제20권10호
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• pp.1436-1439
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• 2010

Many studies have clarified that poly(${\gamma}$-glutamic acid) (PGA) increases the solubility of $Ca^{2+}$, suggesting that PGA enhances calcium absorption in the small intestine. However, there has been no report on the specific interaction between PGA and $Ca^{2+}$ in water. We studied the aqueous solution properties of PGA calcium salt (PGA-Ca complex). The chelating ability and binding strength of PGA for $Ca^{2+}$ were evaluated. The PGA-Ca complex was soluble in water, in contrast to the insolubility of poly(acrylic acid) (PAA) calcium salt, and the chelating ability of PGA for $Ca^{2+}$ was almost the same as that of PAA. The globular conformation of the PGA-Ca complex in water was estimated by SEC and viscosity measurements. The chelation ability of PGA for $Ca^{2+}$ was examined by $^1H$ NMR. The present study showing the characteristics of the PGA-Ca complex will provide useful information about the calcium absorption by PGA in vivo.

• Journal of Microbiology and Biotechnology
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• 제27권5호
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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.