• Title/Summary/Keyword: Poly-${\gamma}$-glutamic acid (${\gamma}$-PGA)

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Synthesis of an Amphiphilic Poly(${\gamma}$-Glutamic Acid)-Cholesterol Conjugate and Its Application as an Artificial Chaperone

  • Lee, Eun-Hye;Kamigaito, Yoshiki;Tsujimoto, Takashi;Uyama, Hiroshi;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.20 no.10
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    • pp.1424-1429
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    • 2010
  • A poly(${\gamma}$-glutamic acid) (${\gamma}$PGA)-cholesterol conjugate was synthesized and its properties were then evaluated. The conjugate exhibited an amphiphilic nature derived from the hydrophilic ${\gamma}$PGA backbone and the hydrophobic cholesterol side chain. The conjugate spontaneously formed nanoparticles, becoming an aqueous solution when at low concentrations, and at high concentrations the result was the formation of a physical gel. By utilizing the self-aggregating properties of the conjugate in water, an artificial chaperone was developed. A complex of protein, with the nanoparticles of the conjugate, was formed and the protein was released upon the dissociation of the nanoparticles through the addition of ${\beta}$-cyclodextrin. For denatured carbonic anhydrase, the activity was recovered in the artificial chaperone of the nanoparticle conjugate.

Isolation of Bacillus sp. Producing Poly-$\gamma$-glutamic Acid with High Efficiency and Its Characterization (고효율 Poly-$\gamma$-Glutamic Acid생산 균주의 분리 및 생산 특성)

  • You Kyung-Ok;Oh You-Na;Kim Byung-Woo;Nam Soo-Wan;Jeon Sung-Jong;Kim Dong-Eun;Kim Young-Man;Kwon Hyun-Ju
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.200-206
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    • 2005
  • A bacterium with high productivity of poly-$\gamma$-glutamic acid (PGA) was isolated from the traditional Korean seasoning, ChungKookJang. The 16s ribosomal RNA sequence of isolated strain showed 97.6, 98.9 and $90.3{\%}$ of similarity to those of Bacillus sp. WL-3, Bacillus subtilis; ENV1 and B amy-loliquefaciens (T), respectively. Accordingly, this bacterium was identified as a Bacillus sp. However, some biochemical characteristics of this strain were different from those of B. subtilis: D-xylose fermentation and glycogen utility were negative. Maximum production of PGA was achieved when it was grown aerobically in a culture medium containing glutamic acid ($3{\%}$) and fructose ($4{\%}$) as carbon sources. The volumetric yield of PGA reached up to 27 g/l in the optimum culture medium. These results suggest that the present strain can be applicable for industrial purposes such as a prototype strain for food or cosmetics industry.

High Molecular Weight Poly-Gamma-Glutamic Acid Regulates Lipid Metabolism in Rats Fed a High-Fat Diet and Humans

  • Park, Ji-Ho;Choi, Jae-Chul;Sung, Moon-Hee;Kang, Jae-Heon;Chang, Moon-Jeong
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.766-775
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    • 2011
  • We investigated the effect of high molecular weight polygamma- glutamic acid (hm ${\gamma}$-PGA) on adiposity and lipid metabolism of rats in the presence of an obesity-inducing diet. Thirty-two Sprague-Dawley rats were fed either a normal-fat (11.4% kcal fat, NFC) or high-fat (51% kcal fat, HFC) diet. After 5 weeks, half of each diet-fed group was treated with hm ${\gamma}$-PGA (NFP or HFP) for 4 weeks. The HFC group had significantly higher body weight, visceral fat mass, fasting serum levels of total cholesterol, LDL cholesterol, and leptin, and lower serum HDL cholesterol level compared with those of the NFC group (p < 0.05). Treatment with hm ${\gamma}$-PGA decreased body weight gain and perirenal fat mass (p<0.05), fasting serum total cholesterol, and mRNA expression of glucose-6- phosphate dehydrogenase (G6PD), regardless of dietary fat contents (p < 0.01). However, hm ${\gamma}$-PGA increased serum HDL cholesterol in the HFC group (p < 0.05). In vitro, 3-hydroxy-3-methylglutaryl coenzyme-A (HMGCoA) reductase activity was suppressed by the addition of hm ${\gamma}$-PGA. In agreement with observations in animal study, the supplementation of hm ${\gamma}$-PGA (150 mg/day) to 20 female subjects in an 8-week double-blind, placebocontrolled study resulted in a tendency to decrease total cholesterol and LDL cholesterol concentrations. We thus conclude that dietary supplementation of hm ${\gamma}$-PGA may act as a hypocholestrolemic agent, secondary to its inhibitor effect on HMG-CoA reductase, and decrease abdominal adiposity by decreasing hepatic lipogenesis. The present study is an important first step in establishing the effect of hm ${\gamma}$-PGA on cholesterol levels in rats and humans.

Fabrication of Poly(${\gamma}$-glutamic acid) Monolith by Thermally Induced Phase Separation and Its Application

  • Park, Sung-Bin;Fujimoto, Takashi;Mizohata, Eiichi;Inoue, Tsuyoshi;Sung, Moon-Hee;Uyama, Hiroshi
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.942-952
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    • 2013
  • Monoliths are functional porous materials with a three-dimensional continuous interconnected pore structure in a single piece. A monolith with uniform shape based on poly(${\gamma}$-glutamic acid) (PGA) has been prepared via a thermally induced phase separation technique using a mixture of dimethyl sulfoxide, water, and ethanol as solvent. The morphology of the obtained monolith was observed by scanning electron microscopy and the surface area of the monolith was evaluated by the Brunauer Emmett Teller method. The effects of fabrication parameters such as the concentration and molecular mass of PGA and the solvent composition have been systematically investigated. The PGA monolith was cross-linked with hexamethylene diisocyanate to produce the water-insoluble monolith. The addition of sodium chloride to the phase separation solvent affected the properties of the cross-linked monolith. The swelling ratio of the cross-linked monolith toward aqueous solutions depended on the buffer pH as well as the monolith fabrication condition. Copper(II) ion was efficiently adsorbed on the cross-linked PGA monolith, and the obtained copper-immobilized monolith showed strong antibacterial activity for Escherichia coli. By combination of the characteristic properties of PGA (e.g., high biocompatibility and biodegradability) and the unique features of monoliths (e.g., through-pore structure, large surface area, and high porosity with small pore size), the PGA monolith possesses large potentials for various industrial applications in the biomedical, environmental, analytical, and separation fields.

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 $80^{\circ}C$.

Transformation and Mutation of Bacillus licheniformis 9945a Producing ${\gamma}-Poly(glutamic\;acid)$ (${\gamma}-Poly(glutamic\;acid)$ 생산성 균주 Bacillus licheniformis 9945a의 형질전환 미 돌연변이 유도)

  • Chung, Wan-Seok;Ko, Young-Hwan
    • Applied Biological Chemistry
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    • v.40 no.3
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    • pp.173-177
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    • 1997
  • Bacillus licheniformis 9945a releases a natural ${\gamma}-poly(glutamic\;acid)({\gamma}-PGA)$ into fermentation broth and shows a mucoid phenotype on the solid agar medium. Transformation of mucoid cells of Bacillus species has not been simple and straightforward. The transpositional activity of Tn10 in B. licheniformis also has not been own either. Thus, a spontaneous non-mucoid derivative of the B. licheniformis was obtained first. Shuttle vector pHV1248 containing mini-Tn10 was introduced into the non-mucoid derivative by the method of protoplast transformation. The resulting transformant was reverted to the wild mucoid phenotype, and then mutated randomly with the mini-transposon by heat induction. Auxotrophs requiring arginine, lysine, or tryptophan were isolated by replica plating method. Southern blotting and DNA-DNA hybridzation analysis showed that these auxotrophs were generated by mini-Tn10 insertion into the chromosomal DNA. This method of transformation and mutation using pHV1248 would be useful for the generation of diverse mutants of B. licheniformis 9945a.(Received January 24,1997; accepted March 10, 1997)

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Preparation of Chitosan/Poly-${\gamma}$-glutamic Acid Nanoparticles and Their Application to Removal of Heavy Metals (키토산/폴리감마글루탐산 나노입자의 제조 및 중금속 제거에의 응용)

  • Sung, Ik-Kyoung;Song, Jae Yong;Kim, Beom Soo
    • Korean Chemical Engineering Research
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    • v.49 no.4
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    • pp.475-479
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    • 2011
  • Chitosan is a natural polymer that has many physicochemical(polycationic, reactive OH and $NH_2$ groups) and biological(bioactive, biocompatible, and biodegradable) properties. In this study, chitosan nanoparticles were prepared using poly-${\gamma}$-glutamic acid(${\gamma}$-PGA) as gelling agent. Nanoparticles were formed by ionic interaction between carboxylic groups in ${\gamma}$-PGA and amino groups in chitosan. Chitosan(0.1~1 g) was dissolved in 100 ml of acetic acid (1% v/v) at room temperature and stirred overnight to ensure a complete solubility. An amount of 0.1 g of ${\gamma}$-PGA was dissolved in 90 ml of distilled water at room temperature. Chitosan solution was dropped through needle into beaker containing ${\gamma}$-PGA solution under gentle stirring at room temperature. The average particle sizes were in the range of 80~300 nm. The prepared chitosan/${\gamma}$-PGA nanoparticles were used to examine their removal of several heavy metal ions($Cd^{2+}$, $Pb^{2+}$, $Zn^{2+}$, $Cu^{2+}$, and $Ni^{2+}$) as adsorbents in aqueous solution. The heavy metal removal capacity of the nanoparticles was in the order of $Cu^{2+}$ > $Pb^{2+}$ > $Cd^{2+}$ > $Ni^{2+}$ > $Zn^{2+}$.

Differences in Manufacturing Process and Quality between Cheonggukjang for Use in the Raw and Cheonggukjang for Stew

  • Seo, Byoung-Joo;Kim, Young-Ho;Kim, Jong-Kyu
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1279-1284
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    • 2008
  • When cheonggukjgang was manufactured using a Bacillus subtilis CH10-1 starter culture, a short-term fermentation for 14-18 hr appeared to be the optimal for the raw cheonggukjang to avoid the formation of a bitter taste and to contain a high concentration of free sugars, whereas a long-term fermentation for more than 4 days was the optimal for the cheonggukjang for stew in order to contain a high concentration of free amino and organic acids, which are responsible for sweet, savory, and bitter tastes present in stewed cheonggukjang, During activation of murine splenic T cells with phytohemagglutinin (PHA), the presence of either poly-$\gamma$-glutamic acid ($\gamma$-PGA) or partially hydrolyzed $\gamma$-PGA resulted in reduction in the level of interferon-$\gamma$ production and enhancement in the level of interleukin-5 production, possibly due to suppression of Th1 activity and augmentation of Th2 activity. Taken together these results indicate that the raw cheonggukjang and the cheonggukjang for stew are different in their quality and taste as well as immunomodulating activity.

High-Molecular-Weight Poly-Gamma-Glutamate Protects Against Hypertriglyceridemic Effects of a High-Fructose Diet in Rat

  • Jeon, Yeong Hui;Kwak, Mi-Sun;Sung, Moon-Hee;Kim, Sun-Hee;Kim, Myung-Hwan;Chang, Moon-Jeong
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.785-793
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    • 2013
  • We studied the effects of 2 different dosages of high-molecular-weight poly-${\gamma}$-glutamic acid (hm ${\gamma}$-PGA) derived from Bacillus subtilis chungkookjang on lipid metabolism in a high-fructose diet-induced hypertriglyceridemic animal model. For 4 weeks, rats were fed either AIN-93 diet (normal control, NC; n = 10) or modified AIN-93 diet in which cornstarch was substituted with 63% fructose (n = 30) to induce hypertriglyceridemia. After 4 weeks, the hypertriglyceridemic rats were treated with daily oral doses of 0 mg (hypertriglyceridemic control, HC), 2.5 mg (hypertriglyceridemic, low hm ${\gamma}$-PGA, HL), or 5 $mg{\cdot}kg{\cdot}bw^{-1}{\cdot}d^{-1}$ (hypertriglyceridemic, high hm ${\gamma}$-PGA, HH) hm ${\gamma}$-PGA for 4 weeks. The HL and HH groups exhibited significantly lower levels of serum triglyceride, total cholesterol, LDL cholesterol, and free fatty acids than the HC group. The administration of hm ${\gamma}$-PGA reduced serum ALT and AST levels. The activities of lipogenic enzymes such as hepatic malic enzyme and glucose-6-phosphate dehydrogenase as well as glucose-6-phosphate dehydrogenase mRNA expression were significantly decreased by hm ${\gamma}$-PGA administration (p < 0.05). These results indicate that hm ${\gamma}$-PGA has an anti-hypertriglyceridemic effect in high-fructose diet-induced hypertriglyceridemic rats.

Fabrication of Poly(γ-glutamic acid) Porous Scaffold for Tissue Engineering Applications (생체조직공학적 응용을 위한 폴리감마글루탐산 다공성 지지제의 제조)

  • Jeon, Hyeon Ae;Lee, Seung Wook;Kwon, Oh Hyeong
    • Journal of the Korean Society of Manufacturing Process Engineers
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    • v.13 no.3
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    • pp.35-41
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    • 2014
  • Poly(g-glutamic acid) (g-PGA) is a very promising biodegradable polymer that is produced by microorganism of Bacillus subtilis. Because g-PGA is water-soluble, anionic, biodegradable, and even edible, its potential applications have been studied from an industrial standpoint. In this study, we fabricated porous g-PGA foams by means of a freeze-solvent extraction method for tissue-engineering applications. Porous g-PGA foams were chemically cross-linked using a hexamethylene diisocyanate solution. An aqueous basic solution was used to neutralize g-PGA foam for cell culturing. During an in vitro cell culture study, it was observed that primary rabbit ear chondrocytes were well at tached and spread over the surface oft hree-dimensional cross-linkedg-PGA foam. From these results, it is concluded that cross-linkedg-PGA foam is aprom is in gmaterial for tissue-engineering applications, especially those pertaining to the regeneration of human cartilage.