• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.3
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• pp.251-257
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• 2006

The components of the media used to elicit the biosynthesis of $poly-{\gamma}-glutamic$ acid $({\gamma}-PGA)$ by Bacillus subtilis ZJU-7 were investigated, particularly the carbon and nitrogen sources Of the 7 carbon sources investigated, sucrose induced the highest rate of ${\gamma}-PGA$ productivity; among the nitrogen sources, tryptone had the best effect for ${\gamma}-PGA$ production. A $2^{6-2}$ fractional factorial design was used to screen factors that influence ${\gamma}-PGA$ production significantly, and a central composite design was finally adopted to formulate the optimal medium. ${\gamma}-PGA$ productivity improved approximately 2-fold when the optimal medium was used compared with the original nonoptimized medium, and volumetric productivity reached a maximum of 58.2 g/L after a 24-h cultivation period.

• Nutrition Research and Practice
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• v.4 no.1
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• pp.23-29
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• 2010

Poly-gamma-glutamic acid ($\gamma$-PGA) is a mucilaginous and biodegradable compound produced by Bacillus subtilis from fermented soybeans, and is found in the traditional Korean soy product, cheongkukjang. This study was carried out to evaluate the effects of $\gamma$-PGA from a food source on the concentration of the neurotransmitter GABA and its metabolic precursor glutamate in diet-induced obese rats. Eight-week old male Sprague-Dawley rats (n=60) were used. The rats were divided into two groups and obesity was induced by providing either a 10% control fat or 45% high fat diet for 5 weeks. The rats were then blocked into 6 groups and supplemented with a 0.1% $\gamma$-PGA diet for 4 weeks. After sacrifice, brain and serum GABA and glutamate concentrations were analyzed by high performance liquid chromatography with fluorometric detection. The rats fed the high fat diet had significantly increased body weights. $\gamma$-PGA supplementation significantly increased serum concentrations of glutamate and GABA in the control fat diet groups while this effect was not found in the high fat groups. In the brain, glutamate concentrations were significantly higher in the $\gamma$-PGA supplemented groups both in rats fed the normal and high fat diets than in the no $\gamma$-PGA controls. GABA concentrations showed the same tendency. The results indicated that $\gamma$-PGA intake increased GABA concentrations in the serum and brain. However, the effects were not shown in obese rats.

• 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.

• Journal of Microbiology and Biotechnology
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• v.28 no.4
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• pp.527-533
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• 2018

This study compared the radioprotective effects of high-molecular-weight poly-gamma-glutamate (${\gamma}-PGA$, average molecular mass 3,000 kDa) and a reduced form of glutathione (GSH, a known radioprotector) on calf thymus DNA damage. The radiation-induced DNA damage was measured on the basis of the decreased fluorescence intensity after binding the DNA with ethidium bromide. All the experiments used $^{60}Co$ gamma radiation at 1,252 Gy, representing 50% DNA damage. When increasing the concentration of ${\gamma}-PGA$ from 0.33 to $1.65{\mu}M$, the DNA protection from radiation-induced damage also increased, with a maximum of 87% protection. Meanwhile, the maximal DNA protection when increasing the concentration of GSH was only 70%. Therefore, ${\gamma}-PGA$ exhibited significant radioprotective effects against gamma irradiation.

• Journal of Microbiology and Biotechnology
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• v.15 no.1
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• pp.59-65
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• 2005

Poly-${\gamma}$-glutamic acid (PGA) is an unusual anionic polypeptide and has great potential as an environmentally and industrially significant biodegradable material. A new ${\gamma}$-PGA producer, Bacillus mesentericus MJM1, with high production capacity was isolated from Korean domestic Chungkuckjang bean paste. It produced ${\gamma}$-PGA at the level of 10 g/l in suitable media. The viscosities of 5% initially extracted mucin and purified ${\gamma}$-PGA solutions were 660 cps and 600 cps, respectively. The produced ${\gamma}$-PGA polymer consisted of 2,000 glutamic acid residues with even proportion of L and D types with molecular mass of about 200- 300 kDa. Bacillus mesentericus MJM1 displayed ${\gamma}$-glutamyltranspeptidase (${\gamma}$-GTP) activity that is known to play a key role in ${\gamma}$-PGA biosynthesis. The ${\gamma}$-GTP coding region was located on the plasmid of 5.8 kb. The plasmid, named pMMH1, is a rolling-circle replication (RCR) plasmid and additionally contained a replication origin and type I signal peptidase (sipP) coding region.

• 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.

• Journal of Microbiology and Biotechnology
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• v.23 no.9
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• pp.1339-1346
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• 2013

Conventional chemotherapeutic regimens often accompany severe side effects and fail to induce complete regression of chemoresistant or relapsing metastatic cancers. The need for establishing more efficacious anticancer strategies led to the development of a combined modality treatment of chemotherapy in conjunction with immunotherapy or radiotherapy. It has been reported that poly-gamma-glutamate (${\gamma}$-PGA), a natural polymer composed of glutamic acids, increases antitumor activity by activating antigen-presenting cells and natural killer (NK) cells. Here, we investigated the antitumor effect of ${\gamma}$-PGA in combination with cyclophosphamide in a murine melanoma model. Whereas cyclophosphamide alone directly triggered apoptosis of tumor cells in vitro, ${\gamma}$-PGA did not show cytotoxicity in tumor cells. Instead, it activated macrophages, as reflected by the upregulation of surface activation markers and the secretion of proinflammatory factors, such as nitric oxide and tumor necrosis factor ${\alpha}$. When the antitumor effects were examined in a mouse model, combined treatment with cyclophosphamide and ${\gamma}$-PGA markedly suppressed tumor growth and metastasis. Notably, ${\gamma}$-PGA treatment dramatically increased the NK cell population in lung tissues, coinciding with decreased metastasis and increased survival. These data collectively suggest that ${\gamma}$-PGA can act as an immunotherapeutic agent that exhibits a synergistic antitumor effect in combination with conventional chemotherapy.

• 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.

• Korean Journal of Food Science and Technology
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• v.37 no.5
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• pp.812-815
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• 2005

Effects of poly-${\gamma}$-glutamate (PGA) on antioxidative characteristics of amino-carbonyl reaction products of glucose were investigated. Rapid browning reaction was observed under heat and alkaline condition (pH 8.2). Browning products were separated by Sephadex G-50, and brown color intensity and electron-donating ability of DPPH (2,2-diphenyl-1-picrylhydrazyl) of each fraction were measured. Fraction-7 (F-7) and -20 (F-20) showed high DPPH scavenging values. UV-VIS absorption spectrum of F-20 was similar to melanoidin peak, and F-7 showed maximum absorption peak at 270 nm. Molecular weight of F-7 was over 35 kDa as determined by SDS-PAGE. Although F-20 could not be measured on SDS-PAGE, its size was smaller than F-7.

• The Korean Journal of Food And Nutrition
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• v.25 no.3
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• pp.677-684
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• 2012

A bacterium strain GS-2 isolated from the Korean traditional seasoning food, Chungkookjang and was determined to produce large amounts of ${\gamma}$-PGA with high productivity when provided with simple nutrients (L-glutamic acid 2.0%, glucose 1.0%, $NH_4Cl$ 0.5%, $KH_2PO_4$ 0.05%, $MgSO_4{\cdot}7H_2O$ 0.01%, pH 7.0). In this study, the culture medium for this strain was optimized for the production of ${\gamma}$-PGA. The Bacillus subtilis GS-2 required supplementation with L-glutamic acid and other nutrients for maximal production of ${\gamma}$-PGA. The optimal culture conditions for ${\gamma}$-PGA production were a 48 hr culture time, a temperature of $33^{\circ}C$ and initial pH of 6.5 by rotary shaking (220 rpm). A maximum ${\gamma}$-PGA production of 31.0 $g/{\ell}$ was obtained with L-glutamic acid (30 $g/{\ell}$), sucrose (the main carbon source, 30 $g/{\ell}$), $NH_4Cl$ (the main nitrogen source, 2.5 $g/{\ell}$), $KH_2PO_4$ (1.5 $g/{\ell}$) and $MgSO_4{\cdot}7H_2O$ (0.15 $g/{\ell}$) in the culture medium.