• Title/Summary/Keyword: Biotin

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Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor

  • Choi, Sun-Hye;Lee, Ra Mi;Cho, Han-Sung;Hwang, Sung Hee;Hwang, Hong-Ik;Rhim, Hyewhon;Kim, Hyoung-Chun;Kim, Do-Geun;Cho, Ik-Hyun;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.46 no.3
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    • pp.348-356
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    • 2022
  • Background: Gintonin is a ginseng-derived exogenous G-protein-coupled lysophosphatidic acid (LPA) receptor ligand. Gintonin exerts its neuronal and non-neuronal in vitro and in vivo effects through LPA receptor subtypes. However, it is unknown whether gintonin can bind to the plasma membrane of cells and can transactivate the epidermal growth factor (EGF) receptor. In the present study, we examined whether gintonin-biotin conjugates directly bound to LPA receptors and transactivated the EGF receptor. Methods: We designed gintonin-biotin conjugates through gintonin biotinylation and examined whether gintonin-biotin conjugate binding sites co-localized with the LPA receptor subtype binding sites. We further examined whether gintonin-biotin transactivated the EGF receptor via LPA receptor regulation via phosphor-EGF and cell migration assays. Results: Gintonin-biotin conjugates elicit [Ca2+]i transient similar to that observed with unbiotinylated gintonin in cultured PC3 cells, suggesting that biotinylation does not affect physiological activity of gintonin. We proved that gintonin-biotin conjugate binding sites co-localized with the LPA1/6 receptor binding sites. Gintonin-biotin binding to the LPA1 receptor transactivates the epidermal growth factor (EGF) receptor through phosphorylation, while the LPA1/3 receptor antagonist, Ki16425, blocked phosphorylation of the EGF receptor. Additionally, an EGF receptor inhibitor AG1478 blocked gintonin-biotin conjugate-mediated cell migration. Conclusions: We observed the binding between ginseng-derived gintonin and the plasma membrane target proteins corresponding to the LPA1/6 receptor subtypes. Moreover, gintonin transactivated EGF receptors via LPA receptor regulation. Our results suggest that gintonin directly binds to the LPA receptor subtypes and transactivates the EGF receptor. It may explain the molecular basis of ginseng physiology/pharmacology in biological systems.

A Study on the Synthesis of Amphiphilic Styrene Copolymers having Functional Groups on the Side Chain (곁사슬에 기능성기를 갖는 양친매성 스티렌 공중합체의 합성에 관한 연구)

  • Lee, Jung-Bock;Kim, Chang-Bae
    • Applied Chemistry for Engineering
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    • v.5 no.5
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    • pp.801-807
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    • 1994
  • Lactose substituted styrene monomer, N-(p-vinylbenzyl)-D-lactonamide(VLA) was prepared by coupling the lactose lactone with p-vinylbenzylamine. The carboxyl group of biotin was activated with N-hydroxysuccinimide in the presence of N, N'-dicyclohexylcarbodiimide. Subsquently, biotin substituted styrene monomer, N-(p-vinylbenzyl)-biotinamide(VBA) was prepared by amidation of the activated biotin with p-vinylbenzylamine. Poly(vinylbenzylactonamide-co-vinylbenzylbiotinamide), p(VLA-co-VBA) were synthesized through radical polymerization from the synthetic monomers(VLA-VBA) by using various mole ratio. The percentages of yield were 67~71%. The copolymers were found amphlphilic which had hydrophilic lactose, hydrophobic vinylbenzyl and biotin site within the structure. IR and $^{13}C-NMR$ analysis on the monomers and copolymer were carried out.

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Enzyme Sensors Modified with Avidin/Biotin Systembased Protein Multilayers

  • Anzai, Jun-Ichi;Du, Xiao-Yan;Hoshi, Tomonori;Suzuki, Yasuhiro;Takeshita, Hiroki;Osa, Tetsuo
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.591-596
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    • 1995
  • Enzyme multilayers composed of avidin and biotin-labeled enzymes were prepared on the surface of electrode, through a strong affinity between avidin and biotin (binding constant: ca $10^{15} M^{-1}$). The enzyme multilayers were useful for the improvement of the performance characteristies of enzyme sensors. The output current of the enzyme sensors depended linearly on the number of enzyme layers deposited. Thus, lactate oxidase (LOx) and alcohol oxidase (AlOx) were deposited after being modified with biotin for constructing enzyme sensors sensitive to L-lactate and ethanol respectively. It was also possible to deposit two different kinds of enzymes successively in a single multilayer. The glucose oxidase (GOx) and ascorbate oxidase (AsOx) were built into a multilayer structure on a Platinum electrode. The GOx, AsOx multilayer-modified electrode was useful for the elimination of ascorbic acid interference of the glucose sensor.

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Effect of Culture Medium Vitamin Concentration of Culture Medium on Ethanol Production in Syngas Fermentation (합성가스 발효에서 배지 내 Vitamin 농도의 에탄올 생산에 대한 영향)

  • Im, Hongrae;An, Taegwang;Park, Soeun;Kim, Young-Kee
    • New & Renewable Energy
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    • v.17 no.3
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    • pp.8-15
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    • 2021
  • In this study, we assessed the effect of vitamin components (such as biotin, thiamine-HCl, and folic acid) on microorganism microbial growth and ethanol production was examined to enhance increase the ethanol concentration in the Clostridium autoethanogenum culture process using syngas as a sole carbon source. Biotin and folic acid concentrations of 0.2, 2, 20, and 100 ㎍/L were used in the culture experiments at 0.2, 2, 20, and 100 ㎍/L concentrations. The maximum ethanol concentrations of 2.81 g/L and 3.12 g/L were obtained by adding at 0.2 ㎍/L biotin and folic acid, respectively. Moreover, Thiaminethiamine--HCl at concentrations of 0.5, 5, 50, and 250 ㎍/L were was examined evaluated to in the culture experiments. The maximum ethanol concentration of 2.84 g/L was observed at 0.5 ㎍/L of thiamine--HCl. As a resultThus, the optimized concentrations of biotin, thiamine--HCl, and folic acid were determined at 0.2, 0.5, and 0.2 ㎍/L, respectively, for enhancing increasing the ethanol production. In conclusion, the maximum ethanol production was obtained by adding the minimal concentration of vitamins in C. autoethanogenum culture.

Effects of Vitamins and Nucleic acids on the mycelial growth and the sclerotial production of Sclerotium rolfsii (Vitamin과 핵산이 Sclerotium rolfsii의 균사생장 및 균핵형성에 미치는 영향)

  • KIM KI CHUNG
    • Korean journal of applied entomology
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    • v.12 no.2
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    • pp.71-78
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    • 1973
  • The study was performed to clear the effects of thiamine, biotin, nicotinic acid, pyridoxine, inositol, deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) on the mycelial growth and the sclerotial production of Sclerotium rolfsii Sacc. isolated from Magnolia kobus. The results are abstracted as follows: 1. Tested fungus was thiamine- deficient and required thiamine 20r/l for maximum growth of mycelia. At higher concentrations than thiamine 20r/l, however, mycelial growth was decreased with increasing the concentrations and was inhibited little less than that of thiamine-free control at 150r/l. 2. The effecfivenesses of the nitrogen sources on the mycelial growth under the thiamine presence were recognized in order of $NH_4NO_3>(NH_4)_2SO_4 >asparagine> KNO_3$, and on the sclerotial production were $KNO_3>NH_4NO_3>asparagine>(NH_4)_2SO_4$. The optimum concentrations of thiamine were about 12r/1 in $KNO_3$, about 16r/1 in asparagine on the growth of mycelia, and were about 8r/l in $KNO_3\;and\; NH_4NO_3,\; 16r/1$ in asparagine on the production of sclerotia. 3. After the organism began to grow, the pH value of cultral filtrate was rapidly dropped down to about 3.5. Hereafter it was slowly fallen down as the growth amount was increased, but was not depreciated below pH 2.2. 4. Nicotinic acid was not effective individually on the mycelial growth and the sclerotial formation of tested fungus without thiamine, but slight effect of it was recognized with thiamine 10r/l, even though maximum growth was shown at 7-10mg/1. Beyond that concentration, however, mycelial growth was rather depressed. 5. When ammonium sulphate or asparagine as the nitrogen sources was used, pyridoxine, biotin and inositol had not any effectivenesses on the mycelial growth and the sclerotial production of examined fungus. 6. In the concentrations of thiamine, biotin, pyridoxine and inositol, as long as thiamine was not added in those, their correlating effects on the growth of the organism were not observed at all. Equivalent or more effects on the mycelial growth were recognized in combinations of thiamin + pyridoxine, thiamine + inositol, thiamine + biotin + pyridoxine, and thiamine + biotin + pyridoxine + inositol compared with thiamino alone, and in combinations of thiamine + biotin and thiamine + biotin + inositol, mycelial growth was inhibited rather than that of thiamine alone. Sclerotial production of those combinations was increased more than that of thiamine alone in dry weight. 7, The little effects of DNA and RNA on the mycelial growth of the organism were recognized compared with the control(DNA-and RNA-free), and RNA was more effective than DNA. Maximum growth of mycelia was observed at RNA 2-6mg/1 and DNA 6mg/l. No effectivenesses on the sclerotial production were recognized in the RNA and DNA. 8. Mycelial growth of the organism was increased with increasing the concentrations of the RNA and the thiamine, that is, the effectiveness of RNA was revealed apparently under presence of thiamine, but was not shown in the sclerotial formation.

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Enzyme-Linked, Biotin-Streptavidin Bacterial-Adhesion Assay for Helicobacter pylori Lectin-Like Interactions with Cultured Cells

  • Murillo, Guzman;Antonia, Maria;Ascencio, Felipe
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.35-39
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    • 2001
  • A simple method for studying the lectin-like interactions between Helicobacter pylori and cultured human epithelial cell lines was developed using an enzyme-linked, biotin-streptavidin bacterial-adhesion assay. The present study suggests that this method is suitable for evaluating the participation of lectin interactions in the adhesion of H. pylori to cultured HeLa S3 and Kato III cells, both fixed and glycosidase-treated cells, as well as assessing glycoconjugated binding inhibition studies. The time-course and dose-dependent kinetics of the biotin-labeled H. pylori adhesion th the formaldehyde-fixed Hela S3 and Kato III cell lines exhibited saturation. In addition, the binding of the biotin-labeled H. pylori to the formaldehyde-fixed cultured cells was partially blocked by pre-incubation with glycoconjugates and polyclonal antibodies against a heparan sulfate binding protein from H. pylori.

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Bioconversion of Isoflavones and the Probiotic Properties of the Electroporated Parent and Subsequent Three Subcultures of Lactobacillus fermentum BT 8219 in Biotin-Soymilk

  • Ewe, Joo-Ann;Wan-Abdullah, Wan-Nadiah;Alias, Abdul Karim;Liong, Min-Tze
    • Journal of Microbiology and Biotechnology
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    • v.22 no.7
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    • pp.947-959
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    • 2012
  • This study was aimed at an evaluation of the potential inheritance of electroporation effects on Lactobacillus fermentum BT 8219 through to three subsequent subcultures, based on their growth, isoflavone bioconversion activities, and probiotic properties, in biotin-supplemented soymilk. Electroporation was seen to cause cell death immediately after treatment, followed by higher growth than the control during fermentation in biotin-soymilk (P<0.05). This was associated with enhanced intracellular and extracellular ${\beta}$-glucosidase specific activity, leading to increased bioconversion of isoflavone glucosides to aglycones (P<0.05). The growing characteristics, enzyme, and isoflavone bioconversion activities of the first, second, and third subcultures of treated cells in biotin-soymilk were similar to the control (P>0.05). Electroporation affected the probiotic properties of parent L. fermentum BT 8219, by reducing its tolerance towards acid (pH 2) and bile, lowering its inhibitory activities against selected pathogens, and reducing its ability for adhesion, when compared with the control (P<0.05). The first, second, and third subcultures of the treated cells showed comparable traits with that of the control (P>0.05), with the exception of their bile tolerance ability, which was inherited to the treated cells of the first and second subcultures (P<0.05). Our results suggest that electroporation could be used to increase the bioactivity of biotin-soymilk via fermentation with probiotic L. fermentum BT 8219, with a view towards the development of functional foods.

Development of a Sandwich Enzyme Immunoassay for Salmon Gonadotropin II. (연어 생식소자극호르몬 II의 Sandwich Enzyme Immunoassay법 개발)

  • KIM Dae-Jung;HAN Chang-Hee;AIDA Katsumi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.1
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    • pp.55-59
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    • 2000
  • A specific and sensitive sandwich enzyme-immunoassay (EIA) using Avidin-Biotin complex was developed for the measurement of GTH II levels in pituitary content and pituitary cell culture medium of the rainbow trout-(Oncorhpchus mykiss). Biotin-salmon GTH II rabbit IgG (sefondary antibody) wai purified by a protein A sepharose affinity chromatography column and that was biotinylated by using Biotin-N-hydroxysuccinimide ofter (BNHS). Non-biotin salmon GTH II rabbit IgG (first antibody) was obtained only through a protein A sepharose affinity chromatography column. The assay was performed by the so-called 'sandwich' method using a microtiter plate, A dose-response curve was obtained between $0.12 to 125 ng/ml$ of salmon GTH II. The displacement curves for pituitary extraction and pituitary cell culture medium of testosterone-treated rainbow trout were Parallel to the standard curie. The intra-assay and inter-assay coefficients of variation (CV) were $8.2{\%} (N=5) and 12.5{\%} (N=6)$, respectively, This assay system was used to measure the amount of GTH II that accumulated in the culture medium of dispersed pituitary cells in testosterone-treated immature rainbow trout, The accumulation was increased with the amount or salmon gonadotropin-releasing hormone. GTH II values determined by the present method were well correlated with those determined by radioimmunoassay. As a result, this assay system was found to be suitable for the measurement of GTH II for pituitary extraction and pituitary culture medium in many salmonid fishes.

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Bio-functionalized Gold Nanoparticles for Surface-Plasmon- Absorption-Based Protein Detection

  • Kim, Wan-Joong;Choi, Soo-Hee;Rho, Young-S.;Yoo, Dong-Jin
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4171-4175
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    • 2011
  • Bio-functionalized gold nanoparticles (AuNPs), which bio-specifically interact with biotin-(strept)avidin, were investigated in this study. AuNPs were functionalized with a synthetically-provided biotin-linked thiol (BLT), which was synthesized by amidation of the active ester of biotin with 2-mercaptoethylamine. The BLT-attached AuNP was bio-specific for streptavidin, making it potentially useful for biosensor applications. To test the bio-specific interactions, the colors, absorption spectra and TEM images were investigated for proteins such as streptavidin, cytochrome C, myoglobin and hemoglobin. The colors and absorption spectra changed when streptavidin was added to the BLT-attached AuNP solution. However, the color and spectra did not change when the other proteins were added to the same solution. These results show that the AuNPs provided a colloidal solution with excellent stability and highly selective absorption characteristics for streptavidin as a target molecule. Proteins were also screened in order to identify a general strategy for the use of optical biosensing proteins based on AuNPs. In addition, TEM images confirmed that streptavidin led the BLT-attached AuNPs to aggregate or precipitate.

Induction of Methanol Tolerance in Rhizopus nigricans Ehrenberg (Rhizopus nigricans Ehrenberg의 Methanol 내성 유도)

  • 김명희;성혜윤;김말남
    • Korean Journal of Microbiology
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    • v.31 no.4
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    • pp.306-311
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    • 1993
  • The effects of methanol. used as a solvent for the hydrophobic substrate progesterone. on the morphology of Rhizopus nigricans and 11$\alpha$-hydroxylation of progesterone was investigated. The methanol tolerance of the 11$\alpha$-hydroxylase system in polyacrylamide immobilized R. nigricans mycelia as well as in free mycelia has been induced by adding various unsaturated fatty acids. biotin and ions into the cultivation medium. Immobilization of the cell seemed to protect the cells from denaturation by methanol. It gave higher reaction rate of progesterone than the free mycelia in the presence of methanol.500 $\mu$g/l of biotin was found to be the most effective induction agent for the methanol tolerance among tested chemicals. R. nixricans cells sustained its enzymatic activity at higher methanol concentrations as a result of accumulation of unsaturated fatty acids. especially oleic acid. in the membrane phospholipid.

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