• Title/Summary/Keyword: Poly ${\gamma}$ glutamic acid

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Concentration of cell-free fermentation broth containing poly (${\gamma}-glutamic acid$) by ultrafiltration

  • Do, Jin-Hwan;Jang, Ho-Nam;Lee, Sang-Yeop
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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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.

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Effect of γ-PGA (Poly-γ-Glutamic Acid) Supplement on Calcium Absorption and Bone Metabolism in Rats (γ-PGA(Poly-γ-glutamic acid) 보충이 흰쥐의 칼슘 흡수율 및 골대사에 미치는 영향)

  • Lee, Min-Sook;Kang, Jung-Il;Kim, Hyun-Su
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.3
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    • pp.255-261
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    • 2006
  • This study was Conducted to investigate the effect of ${\gamma}-PGA\;({\gamma}-poly\;glutamic\;acid)$ on Ca absorption and bone metabolism in rats. Weaned 4-week old male rats were fed Ca-deficient diets for 3 weeks after the adjustment period. Rats were divided into 6 groups and were fed experimental diets for four weeks. Experimental groups were basal (Ca deficient), control (Ca diet: Ca 0.45%), CP1(Ca 0.45%+casein phosphopeptide 1%), PG1(Ca 0.45%+gamma poly glutamic acid 1%), CPG (Ca 0.45%+casein phosphopeptide 1%+gamma poly glutamic acid 1%) and PG3(Ca 0.45%+gamma poly glutamic acid 3%). Though daily Ca intake and food intake of experimental groups showed no significant difference that of control group. The values of fecal Ca excretion and urinary Ca excretion in groups fed ${\gamma}-PGA$ were significantly lower than that in tile control group. The values of Ca absorption in groups fed ${\gamma}-PGA$ were significantly higher than that in the control group. The levels of femur Ca in ${\gamma}-PGA$ supplemented group were significantly increased compared to the control group. Also, breaking force of femur in ${\gamma}-PGA$ supplemented group showed about 40% increase compared to the control group. These results show that ${\gamma}-PGA$ supplement could be helpful to increase Ca absorption as well as to intensify the femur strength and to increase the Ca content of femur in rats.

Physicochemical Properties of Poly-γ-glutamic Acid Produced by a Novel Bacillus subtilis HA Isolated from Cheonggukjang

  • Seo, Ji-Hyun;Kim, Chan-Shick;Lee, Sam-Pin
    • Preventive Nutrition and Food Science
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    • v.13 no.4
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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.

An Unusual Bioconjugate of Glycerol and Poly(${\gamma}$-Glutamic Acid) Produced by Bacillus subtilis C1

  • SHIH ING-LUNG;WU JANE-YII;WU PEI-JEN;SHEN MING-HAU
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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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.

Characteristics of Hydrogel Prepared from Microbial Poly($\gamma$-glutamic acid) by Chemical Crosslinker

  • Park, Jong-Soo;Choi, Seong-Hyun;Choi, Woo-Young;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.48 no.4
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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 $80^{\circ}C$.

Effects of Ultra High Molecular Weight Poly-${\gamma}$-glutamic Acid from Bacillus subtilis (chungkookjang) on Corneal Wound Healing

  • Bae, Sun-Ryang;Park, Chung;Choi, Jae-Chul;Poo, Ha-Ryoung;Kim, Chul-Joong;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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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.

Preparation and Swelling Characteristics of Hydrogel from Microbial Poly(${\gamma}-glutamic acid$) by ${\gamma}$-Irradiation

  • Choi, Seong-Hyun;Whang, Kyung-Sook;Park, Jong-Soo;Choi, Woo-Young;Yoon, Min-Ho
    • Macromolecular Research
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    • v.13 no.4
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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.

Isolation and Characterization of Bacillus spp. with High-Level Productivity of Poly-γ-Glutamic Acid (Poly-γ-Glutamic Acid 고생성 Bacillus spp. 균주의 분리 및 발효특성)

  • Sim, SangHyeob;Park, Hong-Jin;Oh, HyeonHwa;Jeong, Do-Youn;Song, Geun-Seoup;Kim, Young-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.9
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    • pp.1114-1121
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    • 2017
  • Bacillus strains not producing harmful components were isolated from Korean traditional soybean products. Extracellular enzyme activities (amylase, protease, cellulase, and xylanase) of isolated Bacillus strains were measured, and Bacillus strains with high protease activity were selected. The selected 15 strains were identified as Bacillus amyloliquefaciens (10), Bacillus methylotrophicus (1), Bacillus velezensis (1), and Bacillus subtilis (3). Among them, B. subtilis JBG17019, B. amyloliquefaciens JBD17076, and B. amyloliquefaciens JBD17109 showed antimicrobial activities against food-borne microorganisms. The production abilities of glutamate, glutamine, and poly-${\gamma}$-glutamic acid (${\gamma}$-PGA) of the selected Bacillus strains were measured to analyze fermentation characteristics related to glutamic acid metabolism. The factor for multivariate was analyzed by the principal components analysis (PCA) method between fermentation characteristics and ${\gamma}$-PGA production. The three principal components were classified according to the PCA method: PC1 [enzyme activity (amylase, cellulase, and xylanase)], PC2 (${\gamma}$-PGA), and PC3 (protease, glutamate, and glutamine). As a result, B. amyloliquefaciens JBD17076 and B. subtilis JBG17019 strains were evaluated as having excellent enzyme activity and ${\gamma}$-PGA production.

Chelation of Calcium Ions by Poly(${\gamma}$-Glutamic Acid) from Bacillus subtilis (Chungkookjang)

  • Tsujimoto, Takashi;Kimura, Junya;Takeuchi, Yasushi;Uyama, Hiroshi;Park, Chung;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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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.

The Poly-γ-ᴅ-Glutamic Acid Capsule of Bacillus licheniformis, a Surrogate of Bacillus anthracis Capsule Induces Interferon-Gamma Production in NK Cells through Interactions with Macrophages

  • Lee, Hae-Ri;Jeon, Jun Ho;Rhie, Gi-Eun
    • 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.