• 약학회지
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• 제8권3호
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• pp.85-88
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• 1964

Pantothenic acid and biotin contents in Panax Ginseng roots were determined microbiologically with L. arabinosus 17-5. Detection of the vitamins was achieved by the thin-layer chromatography and PPC. Pantothenic acid and biotin were found at the Rf values of 0.42 and 0.55 respectively on the thin-layer chromatograms. In order to find out whether or not the L. arabinosus growth promoting factors contained in the respective ginseng extracts, as shown by the microbiological assay, were really the vitamins respectively, PPC was carried out. Microbiological assay of the vitamins met with results that the average pantothenic acid content in the roots was 6.6r/g, calculated as Ca-pantothenate, and the average biotin content 9.24 mr/g.

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권4호
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• pp.293-300
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• 2001

A polyrotaxane-biotin conjugate was synthesized and its interaction with streptavidin measured using surface plasmon resonance(SPR) detection. A biodegradable polyrotaxane in which ca, 22 molecules of ${\alpha}$-cyclodextrina(${\alpha}$-CDs) were threaded onto a poly(ethylene oxide) chain(M$\sub$n:4,000) capped with benzyloxycarbonyl-L-phenylalanine was conjugated with a biotin hydorazide and 2-aminoethanol after activing the hydroxyl groups of ${\alpha}$-CDs in the polyrotaxane using N, N'-carbonyldiimidazole. The results of the high-resolution $^1$H-nyclear lmagnetic resonance($^1$H-NMR)spectra and gel permeation chromatography of the conjugate showed that ca, 11 biotin molecules were actually introduced to the polyrotaxane scaffold. An SPR analysis showed that the binding curves of the biotin molecules in the conjugate on the streptavidin-deposited surface changed in a concentration dependent manner, indicating that the biotin in the conjugate was ac-tually recognized by streptavidin. The association equilibrium constant(K$\sub$a/) of the interaction be-tween the conjugate and steptavidin tetramer was of the order 10$\^$7/. These results suggest that polyrotaxane is useful for scaffolds as a polymeric ligand in biomedical fields.

• Bulletin of the Korean Chemical Society
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• 제35권2호
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• pp.383-391
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• 2014

Biotin-S-S-Phosphine was designed and synthesized as a potential tool for a proteomic study of O-GlcNAcmodified proteins. This reagent features a disulfide linker between a triarylphosphine moiety, which allows selective conjugation to azide-containing proteins, and a biotin moiety that can allow easy isolation through its strong affinity toward avidin-coated solid beads. The disulfide linkage within this reagent can allow the easy release of the bound molecules of interest, which is difficult to achieve when a biotin:avidin pair is used alone, by reducing the disulfide bond of the reagent with DTT. Preliminary in vitro biological assays with azidelabeled and unlabeled cell lysates and a pure protein Nup62 showed that the Biotin-S-S-Phosphine reagent is highly reactive toward the free thiol groups of proteins. When a molecular tool with a disulfide linker is applied to the enrichment of the molecules of interest from other species, it is important to block the free-thiols of the sample using exhaustive alkylation prior to the Staudinger ligation reactions to restore the bioorthogonal nature of this reaction.

• 미생물학회지
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• 제26권3호
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• pp.149-154
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• 1988

Japanese Encephalitis Virus(JEV) can be detected conveniently by the use of biotinylated cDNA probe. To prepare biotinylated probe aminoallyl dUTP was first synthesized chemically to reverse transcribe the virial RNA. The allylamine-labeled cDNA was then converted to the biotin-cDNA by the reaction with an activated biotin ester, NHS-ACA-biotin. The JEV genomic RNA was hybridized to the biotinylated cDNA probe on nitrocellulose filter and visualized colorimetrically by streptavidin complexes with alkaline phosphatase polymer. Sensitivity of the detection system was determined by estimating the amount of the JEV genomic RNA through comparison with signals generated from the biotinylated and $^{32/P}$ -labeled probes. It was found that the biotin probe was as sensitive as $^{32/P}$ -cDNA probe which can detect 50pgs of the target RNA.

• 센서학회지
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• 제25권5호
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• pp.320-325
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• 2016

In this paper, we propose an AlGaN/GaN-based extended-gate metal-insulator-semiconductor high electron mobility transistor (MISHEMT)-type biosensor for detecting streptavidin-biotin complexes. We measure the drain current of the fabricated sensor, which varies depending on the antibody-antigen reaction of streptavidin with biotin molecules. To confirm the immobilization of biotin polyethylene glycol (PEG) thiol, we analyze the Au surface of a GaN sample using X-ray photoelectron spectroscopy (XPS). The proposed biosensor shows higher sensitivity than Si-based extended-gate metal oxide semiconductor field effect transistor (MOSFET)-type biosensor. In addition, the proposed AlGaN/GaN-based extended-gate MISHEMT-type biosensor exhibits better long-term stability, compared to the conventional AlGaN/GaN-based MISHEMT-type biosensor.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1369-1376
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• 2006

Self-organized nanogels were prepared from pullulan/biotin conjugates (PU/Bio) for the development of an effective anticancer drug delivery system. The degree of biotin substitution was 11, 19, and 24 biotin groups per 100 anhydroglucose units of pullulan. The physicochemical properties of the nanogels (PU/Bio1, 2 and 3) in aqueous media were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of all the samples was less than 300 nm with a unimodal size distribution. The critical aggregation concentrations (CACs) of the nanoparticles in distilled water were $2.8{\times}10^{-2},\;1.6{\times}10^{-2}$, and $0.7{\times}10^{-2}mg/ml$ for the PU/Bio1, 2, and 3, respectively. The aggregation behavior of the nanogels indicated that biotin can perform as a hydrophobic moiety. To observe the specific interaction with a hepatic carcinoma cell line (HepG2), the conjugates were labeled with rhodamine B isothiocyanate (RITC) and their intensities measured using a fluorescence microplate reader. The HepG2 cells treated with the fluorescence-labeled PU/Bio nanoparticles were strongly luminated compared with the control (pullulan). Confocal laser microscopy also confirmed internalization of the PU/Bio nanogels into the cancer cells. Such results demonstrated that the biotin in the conjugate acted as both a hydrophobic moiety for self-assembly and a tumor-targeting moiety for specific interaction with tumor cells. Consequently, PU/Bio nanogels would appear to be a useful drug carrier for the treatment of liver cancer.

• Parasites, Hosts and Diseases
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• 제31권1호
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• pp.37-42
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• 1993

질트리코모나스(Trichomonasvqqin is)의 항원성 변이를 관찰하기 위해 원충의 표면 항원 (surface antigen)을 N-hydmwsuccinlmide-biotin(N반5-biotin)으로 표지 하고 표지된 표면 단백질과 토끼의 항혈청으로 면역침전(Immunoprecipitation; IP)시켰으며 sodium dodec낀 sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)와 전기영동이적법을 시 행하였다. 살아있는 원충을 NHS-biotin으로 표지하여 표면 단백질을 분리하고 이를 질트리코모나스에 면역된 토끼 항혈청과 면역침전 시켰던 바 46, 60, 68, 90, 130 그리고 220 kDa에서 6개의 단백질이 항원성을 나타내었으며 질토리코모나스의 분리주인 HY-1, HV-15 및 ATCC 50148 주간에 차이는 없어 이들 6개 분획이 표면 항원성 발현에 중요한 역할을 할 것으로 생각된다.

• Journal of Ginseng Research
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• 제46권3호
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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.

• 신재생에너지
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• 제17권3호
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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.

• 생명과학회지
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• 제25권6호
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• pp.631-637
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• 2015

Streptmyces avidinii에서 발현되는 Streptavidin은 vitamin H인 d-biotin 4분자에 결합하며 해리상수(Kd)가 10⁻¹⁵ M를 나타내는 아주 강한 비공유결합이다. 이러한 streptavidin과 biotin 상호간의 강한 결합력은 수많은 생물체 분자들의 탐지 및 특징을 연구하는데 응용되어져 왔으므로 Escherichia coli에서 수용성 streptavidin의 기능적 발현에 대한 연구는 매우 유용하다. 즉 Escherichia coli에서 streptavidin을 발현시키기 위해 streptavidin유전자를 T7 RNA polymerase/T7 promoter를 이용하는 pET-22b 플라스미드로 클로닝하였다. 또한 N-말단에 pelB leader를 포함하여 발현된 streptavidin의 periplasmic space로 운반하여 수용성 단백질형태의 분비를 촉진하였으며 C-말단에는 6개의 polyhistidine tags를 두어 정제하는데 사용되었다. 정제된 streptavidin단백질은 10-20 mg/ml 의 높은 회수율을 나타내었으며 SDS-PAGE에서 가열하는 경우 변성되어 17 kD인 monomer형태를, 가열하지 않는 경우에는 68 kDa으로 원래의 tetramer형태를 나타내었다. 따라서 streptavidin의 tetramer 구조는 비공유결합에 의해 이루어짐을 알 수 있었다. Size-exclusion chromatography에 의한 streptavidin의 구조 역시 tetramer를 재확인할 수 있었다. 정제된 수용성 streptavidin은 Westernblot실험에서 biotinylation된 단백질을 탐지하였으며 이 결과는 정제된 streptavidin이 biotin에 결합하는 기능이 존재함을 나타내었다. 이상의 모든 결과를 종합해보면 본 연구에서 구축된 발현시스템을 통하여 발현된 streptavidin은 높은 회수율을 나타내어 대량생산이 가능하였으며 자연상태의 streptavidin과 동일한 homotetramer를 형성하고 biotin에 결합할 수 있는 기능을 나타내었다.