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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李篤中(Duu-Jong Lee) | |
dc.contributor.author | Chung-Yen Shing | en |
dc.contributor.author | 邢忠彥 | zh_TW |
dc.date.accessioned | 2021-06-16T10:13:45Z | - |
dc.date.available | 2015-08-26 | |
dc.date.copyright | 2013-08-26 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-19 | |
dc.identifier.citation | Abou El-Nour, K. M., A. Eftaiha, A. Al-Warthan and R. A. Ammar (2010). 'Synthesis and applications of silver nanoparticles.' Arabian Journal of Chemistry 3(3): 135-140.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60215 | - |
dc.description.abstract | 為發現不同於基於結構設計的新型蛋白酶抑制劑,本研究以自行合成的奈米銀粒子,測試對HIV-1蛋白酶的抑制效果,另外,為提出可能的奈米銀抑制機制,則使用螢光分析技術探討奈米銀與HIV-1蛋白酶在水溶液的交互作用。
實驗結果顯示,以聚乙烯吡咯烷酮(PVP)和檸檬酸根做穩定劑的膠體奈米銀(1-3 nm)皆為良好的蛋白酶抑制劑。在銀/蛋白酶的莫耳比為500、37 ℃、pH 7.0的條件下,與HIV-1蛋白酶控制組相比,PVP穩定跟檸檬酸根穩定的奈米銀能將蛋白酶的酵素最大反應速率(Vmax)由原先的250 μM/min,分別抑制為8.117與2.078 μM/min。由於抑制劑造成HIV-1蛋白酶的米氏常數(Km)與Vmax減低,故奈米銀的抑制類型較有可能屬於反競爭抑制。 在相對活性試驗中,本研究使用檸檬酸根穩定的奈米銀(2, 10, 35 nm),以探討銀粒徑對抑制效果的影響。結果顯示在銀/蛋白酶的莫耳比大於250、37 ℃、pH 7.0的條件下,較小奈米銀(2,10 nm)可抑制HIV-1蛋白酶活性至10%。而對較大奈米銀(35 nm)而言,其抑制活性維持在30%且不隨銀/蛋白酶的莫耳比變化。 螢光分析結果顯示,檸檬酸根穩定的奈米銀是HIV-1蛋白酶的消光劑,其消光類型屬於靜態消光,表示奈米銀與蛋白酶結合、形成一吸收螢光放射的複合物。37 ℃下,奈米銀對蛋白酶的結合位點數為1個。由不同溫度下的結合常數,複合物形成反應的標準焓與熵分別為-96.91 kJ/mol與-170.0 J/mol/K。最後,利用螢光共振能量轉移理論(FRET),可求得奈米銀結合位點與HIV-1蛋白酶上色氨酸的距離,37 ℃下,為5.05 nm。 | zh_TW |
dc.description.abstract | To find a new kind PI different from other structure-based PIs, synthesized silver nanoparticles was examined in their inhibition activities against HIV-1 PR. Besides, the interaction between silver nanoparticles and HIV-1 PR in aqueous solution was also investigated with fluorimetric analysis to propose a possible inhibition mechanism.
The experiment result have shown that both Polyvinylpyrrolidone (PVP)-stabilized and citrate-stabilized colloidal silver nanoparticles (1-3 nm) were good PIs. Comparing with the maximum enzyme reaction rate (Vmax) of HIV-1 PR control (250 μM/min), these two kinds of silver nanoparticles reduced Vmax to 8.117, 2.078 μM/min respectively with Ag/PR molar ratio of 500 at 37 ℃, pH 7.0. The type of both silver nanoparticles inhibition was suggested to be uncompetitive by the reduced Michaelis–Menten constant (Km) and Vmax. Citrate-stabilized silver nanoparticle with different sizes (2, 10, 35 nm) were exploited in relative activity assay of HIV-1 PR to investigate the NPs size effect on the inhibition activity. For smaller silver nanoparticles (2,10 nm), the relative activity of HIV-1 PR was inhibited to ~10% the activity of control with Ag/PR molar ratio greater than 250 at 37 ℃, pH 7.0. For larger silver nanoparticles (35 nm), the relative activity was inhibited to 30% and independent of Ag/PR molar ratio at 37 ℃, pH 7.0. The fluorimetric analysis has shown that the fluorescence quenching of HIV-1 PR by citrate-stabilized silver nanoparticles was static quenching, indicating the silver nanoparticle bind with HIV-1 PR and formed a complex. The number of binding site was equal to 1 at 37 ℃. By using binding constant under different temperatures, the ΔH0 and ΔS0 of binding reaction were calculated to be -96.91 kJ/mol and -170.0 J/mol/K respectively. Finally, based on Fluorescence resonance energy transfer (FRET) theory, the distance between tryptophan residues on HIV-1 PR and silver nanoparticle binding site (37 ℃) is 5.05 nm. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T10:13:45Z (GMT). No. of bitstreams: 1 ntu-102-R99524096-1.pdf: 8316432 bytes, checksum: 5cc837fd20bd2389db0887ce02858d1f (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | ABSTRACT i
摘要 ii ACKNOWLEDGEMENT iii TABLE OF CONTENTS iv LIST OF FIGURES vii LIST OF TABLES ix Chapter 1 Introduction 1 1-1 Background 1 1-2 Research goals 2 Chapter 2 Literature review 3 2-1 Human immunodeficiency virus 3 2-1-1 Current state of HIV prevalence 3 2-1-2 Typing and the virus structure 3 2-1-3 Life cycle of HIV-1 4 2-1-4 HIV-1 protease 5 2-1-5 HIV-1 PR inhibitor 7 2-2 Silver nanoparticles in biomedical application 10 2-2-1 Antimicrobial activities 10 2-2-2 Antiviral activities against HIV-1 11 2-2-3 Medical use 13 2-2-3-1 Impregnation of medical devices 13 2-2-3-2 Anti-inflammatory and wound dressing 13 2-2-4 Effect on and enzyme activity 14 2-3 Ag NP synthesis 16 2-3-1 Mechanism of wet chemical synthesis 16 2-3-2 Reducers in wet chemical synthesis 19 2-3-3 Stabilizers in wet chemical synthesis 19 2-3-4 Green synthesis 21 Chapter 3 Materials and methods 22 3-1 Materials 22 3-1-1 Experimental chemicals 22 3-1-2 HIV-1 PR 23 3-1-3 HIV-1 PR assay kit 23 3-2 Methods 24 3-2-1 Ag NPs synthesis 24 3-2-1-1 PVP-stabilized Ag NPs 24 3-2-1-2 Citrate-stabilized Ag NPs 25 3-2-1-3 Seeding growth of Citrate-stabilized Ag NPs 25 3-2-2 Ag NPs characterization 25 3-2-2-1 Colloidal particle size 25 3-2-2-2 Transmission electron microscopy 26 3-2-2-3 Zeta potential 26 3-2-2-4 UV-Vis spectroscopy 28 3-2-3 HIV-1 PR characterization 28 3-2-3-1 pH measurement 28 3-2-3-2 HIV-1 PR assay 29 3-2-3-3 Calibration 30 3-2-3-4 Kinetic parameters determination 30 3-2-3-5 Stability assay 31 3-2-4 Interaction between Ag NPs and HIV-1 PR 32 3-2-4-1 Inhibition assays 32 3-2-4-2 Ag NPs effect on stability 34 3-2-4-3 Transmission electron microscopy 34 3-2-4-4 Flourimetric analysis 34 3-2-4-5 FRET analysis 36 Chapter 4 Result and discussion 39 4-1 Ag NPs characterization 39 4-2 HV-1 PR characterization 46 4-3 Interaction between Ag NPs and HIV-1 PR 53 Chapter 5 Conclusions 66 REFERENCES 68 | |
dc.language.iso | en | |
dc.title | 奈米銀對HIV-1蛋白酶之抑制效果與交互作用 | zh_TW |
dc.title | Inhibitory Effect and Interaction between Silver Nanoparticle and HIV-1 Protease | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 劉志成(Jhy-Chern Liu),Christopher Whiteley(Christopher Whiteley),黃志彬(Chih-pin Huang),徐治平(Jyh-Ping Hsu),張嘉修(Jo-Shu Chang) | |
dc.subject.keyword | 奈米銀粒子,HIV-1蛋白酶,酵素抑制劑,螢光分析, | zh_TW |
dc.subject.keyword | Silver nanoparticle,HIV-1 protease,Enzyme inhibitior,Fluorimetric analysis, | en |
dc.relation.page | 75 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-08-20 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
顯示於系所單位: | 化學工程學系 |
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