• Title/Summary/Keyword: L. E. M.

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A Study of Organic Acid Formation and Dialysis Culture in E. coli Fermentation (대장균 배양에서의 유기산 생성과 투석배양에 관한 연구)

  • 김인호
    • KSBB Journal
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    • v.8 no.4
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    • pp.382-389
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    • 1993
  • In order to elucidate the effect of acetate ion on the growth of Escherichia coli, flask and fermentor cultures were performed using M9 and LB media. Acetic acid was secreted at a higher rate under the conditions of high glucose concentration as well as of richer medium, i. e., LB broth. The pH in flask culture could not be controlled as i fermentor and pH decreased with the formation of acetic acid. The inhibition effect of acetic acid was pronounced at a lower pH, and the effective inhibitory concentrations of acetic acid were 2.0g/l for LB flask culture, 4.0g/l for M9 flask culture, and 8.0g/l for M9 fermentor culture. Dialysis flask culture was designed to slowly provide E coli cells with glucose. Solid LB agar was layered under LB liquid medium with the variation of agar concentration and solid volume, The increase in the solid portion in the total volume(agar+liquid) resulted in the increase of the final cell concentration. This can be ascribed to the fact that the larger solid phase behaves like a larger reservoir for glucose and controls the growth of E. coli with a controlled rate.

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Structure and Photoreaction of Photoactive Yellow Protein

  • Imamoto, Yasushi;Harigai, Miki;Shimizu, Nobutaka;Kamikubo, Hironari;Yamazaki, Yoichi;Kataoka, Mikio
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.126-129
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    • 2002
  • The chromophore/protein interactions in the photocycle intermediates of photoactive yel- low protein (PYP) were probed by site-directed mutagenesis. The absorption spectra of L- intermediates produced from E46Q, T50V, and R52Q mutants were calculated using the absorption spectra of dark states and difference absorption spectra between L-intermediates and dark states, and compared with that of PYP$\_$L/. The absorption spectrum of R52Q$\_$L/ agreed with that of PYP$\_$L/, but those of E46Q$\_$L/ and T50V$\_$L/ were red-shifted. The effect of these mutations on the absorption spectrum for L-intermediate was comparable to that for the dark state, suggesting that the interaction around the phe-nolic oxygen of the chromophore is conserved in PYP$\_$L/ unlike the crystal structure. On the other hand, we have reported that the absorption spectra of Y 42F$\_$M/, T50V $\_$M/, and R52Q$\_$M/ agreed with that of PYP$\_$M/, but that of E46Q$\_$M/ was red-shifted, suggesting that the hydrogen bond of the chromophore with Glu46 is conserved but that with Tyr42 is broken in PYP$\_$M/. These results suggest that the chromophore inter-acts with Glu46 throughout the photocycle, but never directly interacts with Arg52. This model con- flicts with some of the structural model of PYP intermediates proposed based on the high-resolution X -ray crystallography.

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Anti-Inflammatory Effects of Fermented Milk Supplemented with Heat-Killed Enterococcus faecalis EF-2001 Probiotics (Enterococcus faecalis EF-2001 유산균 사균체 첨가 발효유의 항염증 효과)

  • Kang, Hyo-Jin;Kim, Tae-Woon;Jhoo, Jin-Woo;Kim, Gur-Yoo
    • Journal of Dairy Science and Biotechnology
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    • v.38 no.2
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    • pp.112-120
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    • 2020
  • This study was conducted to verify the physiological activity of heat-killed Enterococcus faecalis EF-2001 probiotics in fermented milk. The anti-inflammatory effects of fermented milk supplemented with different concentrations (0, 100, and 500 ㎍/mL) of E. faecalis EF-2001 were determined using MTT assay and nitric oxide inhibition assay. The MTT assay was performed using RAW 264.7 cells. Results revealed that the rates of cytotoxicity and cell survival decreased significantly with increase in the concentration of heat-killed probiotics (p<0.05). Moreover, fermented milk supplemented with 100 ㎍/mL EF-2001 (EFM1) and the fermented milk supplemented with 500 ㎍/mL EF-2001 (EFM2) exhibited higher nitric oxide inhibition than normal fermented milk (NFM). Additionally, EFM2 significantly reduced the ratio of prostaglandin E2 compared to NFM (p<0.05). In conclusion, the treatment sample showed higher anti-inflammatory activity than NFM. The findings of this study could be used as a basic guideline for manufacturing of NFM supplemented with heat-killed probiotics.

In vitro Glutathione Production using Mixed Cells in an Aerated Slurry Bioreactor (혼합세포를 이용한 Aerated Slurry Bioreactor에서의 in vitro Glutathione 생산)

  • Go, Seong-Yeong;Gu, Yun-Mo
    • KSBB Journal
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    • v.14 no.4
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    • pp.445-451
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    • 1999
  • Glutathione production was carried out using mixed cells of E. coli TG1/pDG7 $\alpha$ and bakers yeast in an Aerated Slurry Bioreactor. Glutathione-producing enzymes were stable for 34 hours, yielding 4.6 mM glutathione in suspension reaction. Glutahione production with high density mixed cells was studied as a function of flow rate in an Aereated Slurry Bioreactor. Glutathione concentration was higher than that in suspension reaction for 32 hours at the substrate feeding rate of 5.2 mL/hr with cell recycle in continuous Aerated Slurry Bioreactor. It was for 42 hours at 2.6 mL/hr and 22 hours at 5.2 mL/hr without cell recycle. Glutahione productivity was 25.7 mg/g wet $cell{\cdot}hr$ at the substrate feeding rate of 10.4 mL/hr with cell recycle, but 5.28 mg/g wet $cell{\cdot}hr$ at 5.2 mL/hr and 1.65 mg/g wet $cell{\cdot}hr$ at 2.6 mL/hr without cell recycle. Effective production time increased from 25 to 45 hours, by using a surfactant, tween 80. As a purfing gas, nitrogen was tested instead of air to avoid a possible oxidizing effect on glutathione-producing enzymes, resulting in the increase of effective production time to 40 hours.

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Studies on the Properties of E. coli ${\gamma}-Glutamylcysteine$ Synthetase in Relation to the Enzymatic Synthesis of Glutathione (글루타치온의 효소적 생합성에 관계되는 E.coli ${\gamma}-Glutamylcysteine$ Synthetase의 특성 연구)

  • Nam, Yong-Suk;Kwak, Joon-Hyeok;Lee, Se-Yong
    • Applied Biological Chemistry
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    • v.40 no.6
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    • pp.478-483
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    • 1997
  • ${\gamma}-Glutamylcysteine$ synthetase was purified from E. coli K-12 strain and its properties related to the in vitro synthesis of glutathione by enzymatic method were investigated. The activity of purified ${\gamma}-glutamylcysteine$ synthetase was increased with increasing concentration of L-glutamate up to 60 mM, while it was decreased by about 50% and 40% under 60 mM of L-cysteine and 45 mM of glycine, respectively. The enzyme activity was reduced not only by ADP, one of the reaction products, but also by the reduced form of glutathione. Therefore, because the reduced glutathione as well as glycine which is the substrate for glutathione synthetase inhibit the activity of ${\gamma}-glutamylcysteine$ synthetase, it is recommended to design a bioreactor system with two separate reactions for glutathione synthesis : one with ${\gamma}-glutamylcysteine$ synthetase reaction and the other glutathione synthetase reaction. In addition since ADP, resulted from these reactions, reduces the activity of ${\gamma}-glutamylcysteine$ synthetase, it is necessary to introduce an ATP regeneration system for glutathione synthesis.

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Antimicrobial Activity of Glycerol Monolaurate and Organic Acids on the Survival of Escherichia coli O157:H7

  • Lee, Myung-Ki;Park, Boo-Kil;Jeong, Cha-Kweon;Oh, Deog-Hwan
    • Preventive Nutrition and Food Science
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    • v.6 no.1
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    • pp.6-9
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    • 2001
  • Minimal inhibitory concentrations (MIC) and antimicrobial effects of glycerol monolaurate (monolaurin) and organic acids, either alone or in combination, against Escherichia coli O157:H7 in tryptic soy broth were determined. MIC values of monolaurin(ML), acetic (AA), citric (CA), lactic (LA) and hydrochloric acid (HCl) were 300$\mu\textrm{g}$/mL (0.03%), 1250$\mu\textrm{g}$/mL(0.125%), 5000$\mu\textrm{g}$/mL(0.5%), 2500$\mu\textrm{g}$/mL(0.25%) and 2500$\mu\textrm{g}$/mL(0.25), respectively. When 150$\mu\textrm{g}$/mL of ML was combined with 50$\mu\textrm{g}$/mL AA, 250$\mu\textrm{g}$/mL HCl and 125$\mu\textrm{g}$/mL LA, the combined agents did not increase the inhibitory effect of the most active single compound alone. This result indicates that there was little interaction between ML and A, HCl and LA. However, the combination of 150$\mu\textrm{g}$/mL ML and 250$\mu\textrm{g}$/mL CA synergistically inhibited growth of E. coli O157:H7. The present study showed that the nature of combined antimicrobial response against E. coli O157:H7 was complex, but this information would be useful for determining interaction that could compromise effectiveness in food systems.

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Antibacterial activity of Dioscorea batatas, Morus alba, and Tagetes erecta against E. coli and S. aureus

  • Hyun-Seo Yoon;Chung Mu Park
    • Biomedical Science Letters
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    • v.29 no.3
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    • pp.130-136
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    • 2023
  • In this study, the anti-bacterial activity of three medicinal herbs such as Dioscorea batatas, Morus alba, and Tagetes erecta, have been applied in oriental medicine against Escherichia coli and Staphylococcus aureus. The antibacterial activity of Dioscorea batatas ethanol extract (DBEE), Morus alba ethanol extract (MAEE), and Tagetes erecta ethanol extract (TEEE) was analyzed by disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) assays. As a result, three medicinal herbs exhibited antibacterial activity in a dose-dependent manner as well as MAEE and TEEE showed the most potent antibacterial activity followed by disk diffusion assay against E. coli and S. aureus, respectively. MIC against E. coli and S. aureus was 1.6 mg/mL of MAEE and TEEE as well as 0.2 mg/mL of TEEE. MBC against both strains was 5 mg/mL of DBEE and 2.5 mg/mL of TEEE. Consequently, three medicinal herbs in this study showed potent antibacterial activity through the inhibited growth of E. coli and S. aureus, which is considered as a potential candidate for cosmeceutic agents to attenuate the pruritus. In addition, antibacterial and antihistamine effects followed by the extraction solvents should be analyzed in future studies.

Effects of 17β-estradiol, Interleukin-1β, and Human Chorionic Gonadotropin on Activity and mRNA Expression of Plasminogen Activators in Porcine Endometrial Cells

  • Hwangbo, Yong;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.22 no.2
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    • pp.155-163
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    • 2018
  • This study aimed to investigate changes in the activity and mRNA expression of plasminogen activators (PAs) induced by $17{\beta}$-estradiol ($E_2$), human chorionic gonadotropin (hCG), and interleukin-$1{\beta}$ ($IL-1{\beta}$) in porcine endometrial cells. Endometrial cells were isolated from the epithelium and cultured to 80% confluence. They were then treated for 24 h with $E_2$ (0.2, 2, 20, and 200 ng/mL), $IL-1{\beta}$ (0.1, 1, 10, and 100 ng/mL), and hCG (0.5, 1, 1.5 and 2 IU/mL). mRNA expressions of urokinase-type (uPA) and tissue-type (tPA) PAs were analyzed using reverse transcription PCR, and activities were measured using a PA activity assay. mRNA expressions of uPA and tPA increased with $E_2$ treatment; however, this was not significant. Similarly, treatment with hCG did not influence the mRNA expressions of PAs. Interestingly, treatment with 0.1 ng/mL $IL-1{\beta}$ significantly reduced the mRNA expression of uPA, but did not affect that of tPA. Treatment with 2, 20, and 200 ng/mL $E_2$ increased PA activity compared with the control group; treatment with 0.1 and 1 ng/mL $IL-1{\beta}$ significantly increased PA activity compared with the other $IL-1{\beta}$ treatment groups, whereas treatment with 10 and 100 ng/mL $IL-1{\beta}$ decreased. Treatment with 2 IU/mL hCG increased PA activity compared with the other treatment groups, although there were no significant differences between the hCG and control groups. In conclusion, the activity and mRNA expression of PAs were differently regulated by the hormone/cytokine and its concentration in porcine endometrial cells. Therefore, understanding PA regulatory mechanisms may help to improve the reproductive potential of domestic animals.

Le bilan synthetique de la typagraphie careenne pendant la dynastie de Chason (조선조 활자주조 및 문헌간행의 종합적 평가)

  • Lee Hee-Jae
    • Journal of the Korean Society for Library and Information Science
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    • v.9
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    • pp.53-74
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    • 1982
  • On admet Jusqu'ici que le XVe siecle est une $\acute{e}poque$ phare de l'histoire de l'imprimerie avec l'apparition de la typographie en Orient et en Occident, $c'est-\acute{a}-dire$ en $Cor\acute{e}e$ sur le Rhim. Cependant, $l'\acute{e}poque$ de l'invention typographique en $Cor\acute{e}e$ doit $\hat{e}tre$ $devanc\acute{e}e$ encore d'un ou deux siecle. Outre cette question, il est $\acute{a}$ noter ici que les caracteres mobiles $m\acute{e}talliques$ Kyemi, fondus en 1403, ont $jou\acute{e}$ un $r\hat{o}le$ $tr\acute{e}s$ important dans l'histoire de la typographie mondiale et que les $caract\acute{e}res$ kabin qui leur $succ\acute{e}derent$ en 1434 menerent la typographie $cor\acute{e}enne$ a son $apog\acute{e}e$. Or, les recherches $cor\acute{e}ennes$ se satifait de la primaute de l'invention en Coree des caracteres mobiles metalliques, mais n'adoptent pas une attitude suffisament critique. Aussi, l'auteur a-toil voulu s'interroger sur la veritable valeur de la typographie de la dynastie de Choson tant au point de vue de l'histoire de l'imprimerie mondiale. En premier lieu, on etudie, comme une chronique, les caracteres mobiles metalliques importants tout au long de la dynastie de Choson sans donner une interpretation nouvelle. C'est dans deux derniers, qu'on essaie de developper ses idees sur la typographie en Coree notamment la question technologique qui devrait etre le fond des recherches. Aussi, on y etudie, la technique de la typographie ancienne en Coree interpretee selon une methode scientifique et moderne et, enfin, le bilan synthetique en cherchant les raisons qui freinerent son developpement en Coree malgre la performance de ses debuts. A l'issue de ces recherches, on ne peut s'empecher d'eprouver une certaine insatisfaction a cause de l'ambiguite des explications que l'on trouve dans des documents fondamentaux. Avec ce difficulte on a voulu, au moins, exposer des points discutatables qui devraient etre les recherches a futur.

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Equine Motor Neuron Disease in a Jeju Pony (제주 조랑말에서의 말운동신경세포질환 일례)

  • Lee, Sang-Kyu;Han, Jae-Ik;Kang, Hyun-Gu
    • Journal of Veterinary Clinics
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    • v.30 no.2
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    • pp.142-145
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    • 2013
  • Equine motor neuron disease (EMND) is a spontaneous neurologic disorder of horses, which results from the degeneration of motor neurons in the spinal cord and brain stem. An 8-year-old Jeju pony gelding presented with weight loss, muscle tremors, frequent recumbence, low head carriage, sweating, and standing with four limbs close together. The gelding has been on the same stable and limited access to pasture for several years. The gelding has been fed with dried hays and commercial concentrated feeds. Laboratory test revealed very low serum vitamin E level (0.14 ${\mu}g/mL$; reference range > 1.5 ${\mu}g/mL$), mildly elevated creatine kinase (402 IU/L; reference range 119-287 IU/L) and aspartate aminotransferase (876 IU/L; reference range 226-336 IU/L). Oral glucose absorption test showed decreased glucose absorption. Histopathologic examination using a biopsy specimen from sacrocaudalis dorsalis medialis muscle revealed atrophic and hypertrophic muscle fiber, centralization of nucleus, degenerating and necrotic muscle fibers. Taken together, the gelding was diagnosed as EMND. After oral vitamin E administrated for 5 weeks, the gelding showed normally improved stance, decreased periods of recumbency, improved head carriage and weight gain except consistent tremors. This is the first report that successfully treated EMND case occurred in a Jeju pony in Korea.