• Title/Summary/Keyword: L-glutamate

Search Result 333, Processing Time 0.032 seconds

Amplification of Glutathione Production in E. coli Cells Using Recombinant DNA Techniques

  • Nam, Yong-Suk;Park, Young-In;Lee, Se-Yong
    • Journal of Microbiology and Biotechnology
    • /
    • v.1 no.3
    • /
    • pp.157-162
    • /
    • 1991
  • Conditions for glutathione production in E. coli cells which possess pGH501 (2 gshI+gshII) were studied. In terms of ATP supply for the glutathione synthesis, two different systems have been constructed and compared. When the acetate kinase reaction of E. coli was used for ATP generation, 20 mM of L-cysteine was completely converted to glutathione by toluene-treated E. coli cells (100 mg/ml) harboring pGH501 within 2 h at $37^{\circ}C$. However, considering the economical aspects, the glycolytic pathway of yeast was chosen as a better system for ATP generation. The optimal concentrations of reactants for glutathione production were determined to be as follows; 80 mM L-glutamate, 20 mM L-cysteine, 20 mM glycine, 20 mM $MgCl_2$, 50 mM potassium phosphate buffer (pH 7.5), 400 mM glucose, polyoxyethylene stearylamine ($5\;\mul/ml$), toluene-treated E. coli HB101/pGH501 (100 mg/ml), and dried yeast cells (400 mg/ml). The conversion ratio of L-cysteine to glutathione was 80% (about 5 mg/ml) under optimal condition within 6 h at $37^{\circ}C$.

  • PDF

Optimization of Medium Composition for Lipopeptide Production from Bacillus subtilis N7 using Response Surface Methodology

  • Luo, Yi;Zhang, Guoyi;Zhu, Zhen;Wang, Xiaohui;Ran, Wei;Shen, Qirong
    • Microbiology and Biotechnology Letters
    • /
    • v.41 no.1
    • /
    • pp.52-59
    • /
    • 2013
  • The nutritional requirements for the maximum production of lipopeptides by Bacillus subtilis N7 (B. subtilis N7) were investigated and optimized using response surface methodology (RSM) under shake flask fermentation. A one-factor-at-a-time experimental setup was used to screen carbon and nitrogen sources. A Plackett-Burman design (PBD) was employed to screen the most critical variables for lipopeptides production amongst ten nutritional elements. The central composite experimental design (CCD) was finally adopted to elucidate the composition of the fermentation medium. Statistical analyses (analysis of variance, ANOVA) of the results showed that KCl, $MnSO_4$ and $FeSO_4{\cdot}6H_2O$ were important components and that their interactions were strong. Lipopeptide production was predicted to reach 709.87 mg/L after a 60 h incubation using an optimum fermentation medium composed of glucose 7.5 g/L, peanut oil 1.25 g/L, $MgSO_4$ 0.37 g/L, $KH_2PO_4$ 0.75 g/L, monosodium glutamate 6.75 g/L, yeast extract and $NH_4Cl$ (5:3 w/w) 10 g/L, KCl 0.16 g/L, $FeSO_4{\cdot}6H_2O$ 0.24 mg/L, $MnSO_4$ 0.76 mg/L, and an initial pH of 7.0. Lipopeptide production ($706.57{\pm}3.70$ mg/L) in the optimized medium confirmed the validity of the predicted model.

The Anti-diabetic Effects of Kocat-D1 on Streptozotocin-Induced Diabetic Rats (Kocat-D1의 streptozotocin으로 유도한 당뇨모델에 대한 항당뇨 활성)

  • Won, Hye-Jin;Lee, Hyun-Sun;Kim, Jong-Tak;Hong, Chung-Oui;Koo, Yun-Chang;Lee, Kwang-Won
    • Korean Journal of Food Science and Technology
    • /
    • v.42 no.2
    • /
    • pp.204-209
    • /
    • 2010
  • This study was conducted to investigate the anti-diabetic activity of Kocat-D1, which is widely used in traditional medicine to treat diabetes in Shandong, China. Sprague Dawley rats (8 weeks of age) were separated into 4 groups: a normal control, streptozotocin (STZ)-induced diabetic rat group (DM control), Kocat-D1-1 (diabetic rat treated with 0.25 g/kg/day hot water extract), and Kocat-D1-2 (diabetic rat treated with 1 g/kg/day hot water extract). After eight weeks of treatment, the fasting blood glucose levels of the Kocat-D1-1 ($334.3{\pm}32.9\;mg/dL$) and Kocat-D1-2 group ($259.5{\pm}35.0\;mg/dL$) were significantly lower when compared to the DM control group ($451{\pm}42.6\;mg/dL$). Furthermore, the levels of glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), albumin and high-density lipoprotein (HDL) cholesterol in the serum of the Kocat-D1-2 group were significantly normalized when compared to the DM control group. However, significant differences were not observed between the Kocat-D1-1 group and the DM control group. Histochemical staining of the liver of the Kocat-D1-2 group revealed no fat accumulation. The insulin level was significantly upregulated in the Kocat-D1-2 group ($0.13{\pm}0.02\;ng/mL$) when compared to the DM control group ($0.05{\pm}0.04\;ng/mL$). The relative volume of $\beta$-cells in the pancreas of the Kocat-D1-2 group ($49.4{\pm}4.2%$) also increased significantly when compared to the DM control group ($12.9{\pm}7.9%$). These results suggest that Kocat-D1 exerts an anti-hyperglycemic effect through the enhancement of insulin secretion.

In vitro screening of the acetylcholinesterase inhibition, antioxidant activity, and neuronal cell protective effect of medicinal plant extracts (생약추출물의 acetylcholinesterase 저해, 항산화 및 신경세포보호 효과 in vitro 탐색)

  • Um, Min Young;Ha, Tae Youl;Seong, Ki Seung;Kim, Yong Sik
    • Food Science and Preservation
    • /
    • v.20 no.6
    • /
    • pp.840-845
    • /
    • 2013
  • This study investigated the effects (i.e., the acetylcholinesterase activity, lipid peroxidation, and neuronal survival) of 20 kinds of medicinal water extracts. The water extracts of three medicinal plants - Cornus officinalis, Glycyrrhiza glabra, and Angelica gigas - were found to be the most effective on acetylcholinesterase inhibitory activity. In the lipid peroxidation-generating system induced by $H_2O_2/FeSO_4$ in rat brain homogenates, Perilla frutescens, Polygonum multiflorum, Cinnamomun cassia, and G. glabra exhibited protective activity against lipid peroxidation. The neuronal cell death induced by L-glutamate in PC12 was suppressed by the water extracts of G. glabra, Cinnamomun cassia, Platycodon grandiflorum, and Mentha arvensis at the concentration of $100{\mu}g/mL$. Taken together, these results showed that the water extract of G. glabra has the potential anti-dementia activity, which suggests that it might provide an effective strategy for improving dementia.

Serum Steroid Hormone Level and Hematological Characteristics of One-year Cultured eels, Anguilla japonica Based on Total Length and Sex (양식 1년산 뱀장어, Anguilla japonica의 전장 및 성별에 따른 혈중 성호르몬 수준 및 혈액학적 성상)

  • 전민지;한경민;배준영;유진형;이계안;배승철
    • Journal of Aquaculture
    • /
    • v.16 no.4
    • /
    • pp.267-272
    • /
    • 2003
  • This study investigated sex determination through serum steroid hormone level and compared hematological characteristics of one-year old cultured eels, Anguilla japonica. Four different groups were divided with total length at 40∼45, 45∼50, 50∼55, and 55∼60 cm. Males dominated eels of 40∼50 cm in total length group, while females dominated eels of more than 50 cm. All females and males were immature. Hematocrit levels of males (total length: 40∼50 cm) were higher than those of females (total length: 50∼60 cm). Hemoglobin content also differed between males and females. Glutamate oxaloacetate transaminase (GOT) levels of 40∼45 cm males tended to be higher (157$\pm$3.46 IU/L) than that of other sized males, but there were no significant differences between groups (P<0.05). The highest GOT levels were found in of 55∼60 cm females (148$\pm$3.46 IU/L) compared with that of other female groups (P<0.05). Glutamate pyruvate transaminase (GPT) levels of males and females in the 55∼60 cm size range was significantly higher thanothers (P<0.05). The result of this study indicated steroid hormone content of males and females were very low, but testosterone (T) and estradiol-17$\beta$, (E2) contents of females were higher than those of males. For further research of sex determination in cultured eels, it needs to investigate in more defined body length.

Analysis of Secreted Nectar Volume, Sugar and Amino Acid Content in Male and Female Flower of Evodia daniellii Hemsl (밀원수종 쉬나무 수꽃과 암꽃의 화밀분비량, 당 함량 및 아미노산 분석)

  • Kim, Moon Sup;Kim, Sea Hyun;Song, Jeong Ho;Kim, Hyeu Soo
    • Journal of Korean Society of Forest Science
    • /
    • v.103 no.1
    • /
    • pp.43-50
    • /
    • 2014
  • The purpose of study was to analyze secreted nectar volume, nectar sugar content and amino acid in addition to estimating honey quantities that can ultimately reap in male and female flowers of Evodia daniellii Hemsl.. The maximum blooming period of male flowers was on 24 to 26 July in 2012. On average, nectar volume secreted by nectary was $2.73{\pm}0.73{\mu}L$ from one male flower and nectar concentration showed 17.4%. The maximum blooming period of female flowers was on 7 to 9 August in 2012. Nectar volume secreted by nectary was $0.63{\pm}0.49{\mu}L$ from one female flower and nectar concentration showed 25.7%, averagely. As results of correlation analysis between the meteorological factors and nectar characteristics, we found that nectar quantities and concentration were influenced by temperature and relative humidity. Sugar content was calculated at $48.0{\pm}5.2{\mu}g$ per a male flower and $37.8{\pm}8.7{\mu}g$ per a female flower, which meant that both values were not significantly different (Mann-Whitney's U-test, p=0.400). The minimum estimates of honey harvest for a male and female inflorescence were 67.8 g and 53.5 g, respectively. Analysis of amino acid showed that Serine, Glycine and Alanine were more abundant in male flowers, however Asparatate, Glutamate, Asparagine and Glutamine were more abundant in female flowers.

Amino Acid Composition and characteristic of dissolved organic Compounds in the Yellow Sea (황해의 용존 유기물 특성 및 아미노산 조성)

  • 박용철;윤철호
    • 한국해양학회지
    • /
    • v.29 no.2
    • /
    • pp.171-182
    • /
    • 1994
  • Dissolved free amino acid (DFAA) dissolved hydrolyzable amino acid (DHAA) and D/L amino acid racemic ratio in the dissolved organic compounds were studied to investigate the biogeochemical characteristics of dissolved organic compound in the Yellow Sea. Concentration of total DFAA ranged from 0.06 uM to 0.26 uM in the study area. DFAA composition showed that aspiratae, glutamate, serine, glycine and alanine were predominant. According to characteristics of functional group of amino acid, these belonged to hydroponic group. C-18 short column cartridge (Sep-Pak) activated by methanol was used to extract organic macromolecules in the seawater. In operational scheme, macromolecules were divided into two fractions. Geomacromolecule fraction eluted with 50% methanol was used to extract organic macromolecules in the seawater. In operational scheme, macromolecules were divided into two fractions. Geomacromolecule fraction eluted with 50% methanol was moderately hydrophilic and showed characteristics of humic substance in the seawater. Biomacromolecule fraction eluted with 100% methanol was hydrophobic and most abundant in the surface seawater samples. DHAA was much higher than DFAA in this study area. DHAA ranged from 2.05 uM to 6.19 uM in the B-fraction and from 8.13 uM to 24.46 uM in the G-fraction. DHAA was higher in the surface water than in the bottom water where the vertical stratification developed well. The result of HPLC analysis of D/L amino acid showed that low racemic ratio was found in the B-fraction. It implies that the B-fraction is relatively younger than the G-fraction and freshly derived from biosphere.

  • PDF

On-Line Monitoring of Low Biochemical Oxygen Demand Through Continuous Operation of a Mediator-Less Microbial Fuel Cell

  • MOON, HYUN-SOO;CHANG, IN-SEO;JANG, JAE-KYUNG;KIM, KYUNG-SHIK;LEE, JI-YOUNG;LOVITT, ROBERT W.;KIM, BYUNG-HONG
    • Journal of Microbiology and Biotechnology
    • /
    • v.15 no.1
    • /
    • pp.192-196
    • /
    • 2005
  • Abstract Oligotrophic microbial fuel cells (MFCs) were tested for the continuous monitoring of low biochemical oxygen demand (BOD) by using artificial wastewater, containing glucose and glutamate, as check solution. Ten times diluted trace mineral solution was used to minimize the background current level, which is generated from the oxidation of nitrilotriacetate used as a chelating agent. The feeding rate of 0.53 ml/min could increase the sensitivity from 0.16 to 0.43 ${\mu}$A/(mg BOD/l) at 0.15 ml/min. The dynamic linear range of the calibration curve was between 2.0 and 10.0 mg BOD/l, and the response time to the change of 2 mg BOD/l was about 60 min. The current signal from an oligotroph-type MFCs increased with the increase in salts concentration, and the salt effect could be eliminated by 50 mM phosphate buffer.

Rat Liver $\beta$-Glucuronidase; Its Purification and Inhibition Studies

  • Jeong, Han-Seung;Yang, Chul-Hak
    • Bulletin of the Korean Chemical Society
    • /
    • v.6 no.5
    • /
    • pp.312-317
    • /
    • 1985
  • ${\beta}$-Glucuronidase (EC 3.2.1.31) which hydrolizes D-glucuronate from ${\beta}$-D-glucuronide was purified from rat liver, using ammonium sulfate fractionation, DEAE-cellulose chromatography, Concanavalin-A Sepharose 4B chromatography and gel filtration on Sephadex G-200. This enzyme has the molecular weight of 280,000 daltons by gel filtration and 75,000 daltons by SDS-polyacrylamide gel electrophoresis. As its funtion is reverse of detoxification in the liver, the inhibition of the enzyme was tested with extracts of several food products and medicinal herbs, some are known as anti-cancer agents. Among them, Panax ginseng and Cortnellus shiiake inhibited the enzyme competitively and the $K_1$ values were $9.22 {\times}\;10^{-2}$ and 0.102 mg/ml, respectively. These inhibitors strongly bound to DEAE-cellulose. The negatively charged amino acids, L-aspartate and L-glutamate, inhibited the enzyme, and $K_1$ value of L-aspartate was 0.80 mM. The interaction between ${\beta}$-glucuronidase and p-nitrophenyl-${\beta}$-D-glucuronide was found to involve ionic forces by the effect of ionic strength on the kinetic constant, Vmax/Km. It was inferred from these findings that cationic group at the active center of the enzyme is probably involved in attacking the substrate.

Effects of Pueraria radix in Water Extract on the Detoxification in Rat administered with Cadmium (카드뮴을 급여한 흰쥐에서 갈근 열수 추출액의 해독작용효과)

  • Chung, Yung-Hee;Shin, Mee-Kyung;Han, Sung-Hee
    • Journal of the Korean Society of Food Culture
    • /
    • v.17 no.4
    • /
    • pp.456-464
    • /
    • 2002
  • This study was designed to investigates the effects of Korean pueraris radix water extract in Cd(cadmium) administered rats. Forty male Sprague-Dawley rats weighing $100{\pm}10g$ were used for this experiment and divided into following 4 groups; control group, 3% pueraria radix in water extract group, 50 ppm Cd group, 50ppm Cd group with 3% pueraria radix in water extract group. The Cd administered rats were given 50 ppm of $CdCl_2\;{\cdot}\;2H_2O$ disolved in the distilled water. The Cd content in the rats tissue of Cd administered group was lower than in the rats tissue of Cd group with 3% pueraria radix in water extract group. Plasma levels of renin activity was increased by Cd administration group, compared with 3% pueraria radix in water extract group and Cd administred group. Glutamate oxaloacetate transaminase(GOT) and Glutamate pyruvate transaminase(GPT) were increased in Cd-administered group and lower in the 3% extracts of pueraria radix in water extract group. Lactate dehydrogenase(LDHase) was lower in the 3% extracts of pueraria radix-Cd group than in the Cd group. This results suggested that pueraria radix in water extract group, has a lowering effects on the accumulation of Cd and it is belived that the pueraria radix in water extract group has some protective effects to Cd administered in rats, but the mechanism of these effects was obscure.