• Title/Summary/Keyword: PGA

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An Efficient Method for the Release of Recombinant Penicillin G Amidase from the Escherichia coli Periplasm (대장균의 periplasm으로부터 재조합 PGA 단백질의 효율적이고 간단한 방출 방법)

  • Lee, Sang-Mahn
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1145-1151
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    • 2017
  • In this study, we report on a simple, efficient method for obtaining penicillin G amidase (PGA) from recombinant Escherichia coli using a formulation mixed with detergent and lysozyme. Research was conducted on the extraction efficiency of PGA from the periplasmic space in cells in terms of the type of detergent, detergent concentration, pH, reaction time, and temperature of permeabilization. The extraction yield of PGA in the formulated surfactant/lysozyme treatment was increased by approximately (55-65 U/ml) in comparison with that in the single surfactant treatment. The released PGA solution was concentrated and exchanged with buffer using an ultrafiltration (U/F) system. The yields of diatomite filtration, membrane filtration (M/F), and U/F were 69.7%, 93.8%, and 77.3%, respectively. A total of 212 KU of PGA was recovered. At the 25-L culture scale, the overall yield of extraction using the mixed surfactant/lysozyme method was 49.2%. The specific activity of extracted PGA was 11 U/mg in protein. The concentrated PGA solution was immobilized on microporous silica beads without further purification of PGA. The total immobilization yield of PGA on the resin was 48.7%, while the enzyme activity was 101 U/g. The immobilized PGA was successfully used to produce 6-APA from penicillin G. Our results indicated that a simple extraction method from periplasmic space in E. coli may be used for the commercial scale production of ${\beta}-lactam$ antibiotics using immobilized PGA.

Effects of PGA-LM on CD4+CD25+foxp3+ Treg Cell Activation in Isolated CD4+ T Cells in NC/Nga Mice (NC/Nga 생쥐에서 분리한 T 세포에서 foxp3+ 세포 활성화에 대한 PGA-LM의 효과)

  • Jang, Soon-Nam;Kim, Kum-Lan;Kang, Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.160-169
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    • 2009
  • Poly-$\gamma$-glutamic acid ($\gamma$-PGA) was mixed natural flora of Bacillus subtilis, contaminated from cooked soybeans. Also, it was performed to find out the antiallergic activity by using NC/Nga mice, in vitro. The $\gamma$-PGA (PGA-HM : PGA-high molecular weight), Molecular weight 300 kDa, was decomposed and made PGA-LM (PGA-low molecular weight) which has molecular weight below 30 kDa by sonication. Therefore, it was same result between PGA-HM and PGA-LM, and reported PGA-LM as basic result. We found that PGA-LM contains antiallergic efficacy that inhibit B cells and Th2 cells activation from isolated CD4+T cells in NC/Nga atopic dermatitis model mice, and not show a cytotoxicity in the hFCs. To investigate the effects of these PGA-LM in vitro, isolation of splenic B cell and CD4+ T cells in atopic dermatitis mice were used. To elucidate the role of PGA-LM in anti-CD40+ interleukin-4 (IL-4)-mediated B-cell activation, showed that the capacity of B cells to expression IL-$1\beta$, IL-6, and TNF-$\alpha$ mRNA down-regulated, and IL-10 mRNA up-regulation by PGA-LM treatment, but it had no effect on TGF-$\beta$ expression. In addition to CD4+IFN-$\gamma$+ and CD4+CD25+foxp3+, the functions of PGA-LM in the development of the CD4+CD25+foxp3+ and CD4+IFN-$\gamma$+cells, the phenotype and functions of PGA-LM induced CD4+CD25+foxp3+, and CD4+IFN-$\gamma$+cells in CD4+T cells. These results suggested that PGA-LM could change cytokine production and generate CD4+CD25+foxp3+ Tregs in NC/Nga mice, and may be effective for immunotherapy in patients with AD.

PGA Implementation Technique for Stripmap SAR Signal Processing (Stripmap SAR 신호처리를 위한 PGA 적용 기법)

  • Yoon, Sang-Ho;Koh, Bo-Yeon;Kong, Young-Kyun;Shin, Hee-Sub
    • Korean Journal of Remote Sensing
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    • v.27 no.2
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    • pp.151-161
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    • 2011
  • PGA(Phase Gradient Autofocus) is a representative autofocus technique to improve the SAR(Synthetic Aperture Radar) image quality. PGA can estimate high order phase errors and have good robustness in noisy environments. However, PGA is not suitable to apply to the stripmap mode data directly because it is based on the spotlight mode operation. In this paper, the PGA implementation technique for stripmap mode data and the method of ROI(Region of Interest) selection that affects severely on PGA performance have been proposed. The proposed technique was verified by the point target simulation first, and was applied to the real SAR signal data acquired by the flight test. Finally, the significant improvements in focusing quality were shown in the processed SAR images using the proposed method.

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.

Degradation Behavior of Medical Resorbable Composite Materials Interposed in the Poly(glycolic acid) (Poly(glycolic acid)를 심선에 지닌 의료용 흡수성 복합재료의 생분해 거동)

  • Lee, Chan-Woo
    • Polymer(Korea)
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    • v.31 no.3
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    • pp.233-238
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    • 2007
  • The purpose of the study is to apply composites of poly (glycolic acid) (PGA) with [poly(R) 3-hydroxybutyrate] (P3HB) or poly (butylenes succinate- co-L-lactate) (PBSL) as medical resorbable composite materials with the complement of hydrolysis rate of each component. As a result, it was confirmed that the PBSL/PGA and P3HB/PGA composite fiber were hydrolyzed in phosphate buffer solution. Also, it has been revealed that the degradation of PBSL/PGA are accelerated due to PGA producing glycolic acid which can act as a catalyst. In addition, the hydrolysis of PBSL/PGA was found to be accelerated by the presence of lipase PS. When the PBSL/PGA composite fiber was placed in the air, not much hydrolysis has proceeded. Also, it was confirmed that the P3HB/PGA composite fiber maintained proper tensile strength in the air. Therefore, these complex fibers can be adapted to use as environmentally suitable, medically absorbable composite materials.

The Effect of $\gamma$-PGA on NC/Nga Mice, a Mouse Model for Mite Antigen-induced Atopic Dermatitis (집먼지 진드기 항원으로 아토피 피부염을 유발한 NC/Nga 생쥐에 미치는 $\gamma$-PGA의 효과)

  • Jang, Soon-Nam;Kim, Kum-Lan;Yun, Mi-Young;Kang, Sang-Mo
    • Microbiology and Biotechnology Letters
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    • v.38 no.1
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    • pp.53-63
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    • 2010
  • As one of the mucous components of Cheonggukjang, traditional fermented soybean paste, $\gamma$-PGA is a natural substance with diverse functions. In this paper, an in-vivo experiment has been performed using NC/Nga mice in order to find out the efficacy of $\gamma$-PGA in human atopic dermatitis. The NC/Nga mice with BMAC-induced atopic dermatitis were administered $\gamma$-PGA (PGA-HM) with 300 kDa and low-molecular $\gamma$-PGA (PGA-LM), respectively. As a result, a significant decrease in clinical skin severity score was detected in the group that was administered PGA-LM. In terms of serum IgE levels, a significant decline was observed in PGA-LM, compared to the control group. The serum IgG1 levels also decreased more in PGA-LM than in the control group. However, no significant difference was observed in both groups. To witness the induction of $CD4^+CD25^+foxp3^+$ Treg cells, mRNA was sampled from the back of PGA-HM- and PGA-LM-administered NC/Nga mice with atopic dermatitis. In terms of the production amount of foxp3 mRNA, which was measured in real-time PCR, the group that was administered PGA-LM was twice as high as the control group. According to a biopsy on the skin on the backs of the mice, the experimental group was also far lower than the control group in terms of epidermis thickness, mast cell infiltration and the number of $CCR3^+$ cells. Therefore, it has been confirmed that the atopic dermatitis symptoms decreased more in the PGA-LM-administered NC/Nga mice than the PGA-HM-administered group by facilitating $CD4^+CD25^+foxp3^+$ Treg cells and suppressing the activity of eosinophils and production of IgE and pro-inflammatory cytokines.

Poly(D-glutamic acid)-Fe(II) Complex Formation and Its Stability Constant (Poly(D-glutamic acid)-Fe(II) 복합물의 형성과 안정도 상수에 관하여)

  • Chong-Su Cho;Seon-Ung Kim
    • Journal of the Korean Chemical Society
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    • v.31 no.5
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    • pp.471-475
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    • 1987
  • Fe(II)-poly(D-glutamic acid) (PGA) complex in aqueous solution has been investigated as functions of pH and concentration of Fe(II) by using absorption and circular dichroism spectroscopies. The helix-breaking interaction occurred at pH 4.3 while the helix-directing one occurred at pH 5.7 after forming complex between PGA and Fe(II). The relationship between conformation change of PGA by forming complex with Fe(II) and complex constants for the above system was studied. As a result, stability constants in random coil structure of PGA are larger than those in ${\alpha}$-helix structure. This result indicates that the random coil structure of PGA-Fe(II) complex shows more stable complex than ${\alpha}$-helix one.

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The Coagulation Characteristics of Wastewater Using Poly-γ-glutamic Acid (Poly-γ-glutamic acid(PGA)를 이용한 폐수의 응집특성)

  • Kwon, Kwi-bock;Kim, Dong-ha;Kang, Seon-Hong;Sung, Moon-Hee;Park, Chung
    • Journal of Korean Society of Water and Wastewater
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    • v.19 no.3
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    • pp.357-362
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    • 2005
  • Poly-${\gamma}$-glutamic acid (${\gamma}-PGA$), which is extracted from fermented soybeans, is a high molecular weight, adhesive, and negatively charged(anionic) polymer. Recently, ${\gamma}-PGA$ has gained attention due to its potential as polymer. The objectives of this study were to examine the applicability of ${\gamma}-PGA$ as a coagulant and/or a coagulant aid, to evaluate the efficiency of ${\gamma}-PGA$ for the removal of Organic and Ammonium substance in wastewater treatment. The effect of coagulation was evaluated for the removal of SS and organic matter using poly aluminum chloride(PACI) as well as newly developed ${\gamma}-PGA$. The maximum COD removal rate of 63% and the SS of 78% were occurred at the dosage of 50mg/L ${\gamma}-PGA$ only. The most effective removal for particulate and organic matter was occured when both PACI and ${\gamma}-PGA$ were applied at the rate of 20:1(10mg/L PACI and 0.5mg/L ${\gamma}-PGA$). When mixed with PACI, only small portion of ${\gamma}-PGA$ was enough to improve removal efficiencies of organic and particulate matter in wastewater. This result showed the positive potential of ${\gamma}-PGA$ as a new coagulant materials for wastewater treatment.

Fed-batch Culture of Recombinant E.coli for the Production of Penicillin G Amidase (Penicillin G Amidase생산을 위한 재조합 대장균의 유가배양에 관한 연구)

  • Lee, Sang-Mahn
    • Microbiology and Biotechnology Letters
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    • v.36 no.4
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    • pp.314-319
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    • 2008
  • Penicillin G amidase (PGA, benzylpenicillinaminohydrolase, EC 3.5.1.11) is industrially important enzyme which converts penicillin G to 6-aminopenicillanic acid (6-APA) and phenylacetic acid (PAA). The PGA in E. coli ATCC 11105 is secreted into the periplasm after removing signal sequences and becomes heterodimer which composed of two subunits, small subunit (24 kDa) and large subunit (65 kDa). In this study, the PGA gene was obtained from E. coli ATCC 11105 using PCR (polymerase chain reaction) technique. The active PGA was successfully secreated into periplasm in E. coli BL2 1(DE3) harboring pET-pga plasmid. The optimized fed-batch fermentation, consisting of a three-step shift of culture temperature from $37^{\circ}C$ to $22^{\circ}C$, gave a productivity of 19.6 U/mL with a cell growth of 62 O.D. at 600 nm.

Clinical Impact of Polyglycolic Acid Mesh to Reduce Pancreas-Related Complications After Minimally Invasive Surgery for Gastric Cancer: A Propensity Score Matching Analysis

  • Motonari Ri;Manabu Ohashi;Rie Makuuchi;Masaru Hayami;Takeshi Sano;Souya Nunobe
    • Journal of Gastric Cancer
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    • v.24 no.2
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    • pp.220-230
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    • 2024
  • Purpose: Prevention of pancreas-related complications after gastric cancer surgery is critical. Polyglycolic acid (PGA) mesh reduces postoperative pancreatic fistula formation following pancreatic resection. However, the clinical efficacy of PGA mesh in gastric cancer surgery has not been adequately investigated. Materials and Methods: This retrospective study compared the short-term outcomes between two groups: patients who underwent minimally invasive R0 gastrectomy for gastric cancer with the use of a PGA mesh (PGA group) and those without the use of a PGA mesh (non-PGA group) at the Cancer Institute Hospital, Tokyo, between January 2019 and May 2023. Propensity score matching (PSM) was performed to adjust for the possible confounding factors. Results: A total of 834 patients were initially included, of whom 614 (307 in each group) remained after PSM. The amylase levels in the drained abdominal fluid on postoperative days 1 and 3 were similar between the PGA and non-PGA groups. The PGA group had a significantly lower incidence of pancreas-related complications of Clavien-Dindo grade ≥2 than that in the non-PGA group (6.8% vs. 2.9%, P=0.025). In subgroup analyses, the odds ratio for pancreas-related complications appeared to be better in the PGA group than in the non-PGA group in patients with American Society of Anesthesiologists Physical Status Classification score of 2 or 3, those operated via a laparoscopic approach, and those undergoing procedures other than proximal gastrectomy. Conclusions: The use of PGA mesh significantly reduced pancreas-related complications after minimally invasive surgery for gastric cancer and might thus benefit patients at risk of such complications.