Purification and characterization of defense-related proteins from Hokkaido large black soybean and emperor banana.

• Other Authors: Ho, Sai Man.
• Format: Others
• Language: English; Chinese
• Published: 2007
• Subjects: Plant lectins > Purification; Trypsin inhibitors > Purification; Chitinase > Purification; Thaumatins > Purification; Soybean; Bananas; Plant Lectins > isolation & purification; Trypsin Inhibitors > isolation & purification; Chitinase > isolation & purification; Plant Proteins > isolation & purification; Soybeans > chemistry; Musa > chemistry
• Online Access: http://library.cuhk.edu.hk/record=b5893274; http://repository.lib.cuhk.edu.hk/en/item/cuhk-326059

Ho, Sai Man. === Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. === Includes bibliographical references (leaves 144-164). === Abstracts in English and Chinese. === TABLE OF CONTENTS --- p.ii === ABSTRACT --- p.xii === 撮要 --- p.xv === LIST OF ABBREIVIATIONS --- p.xvi === LIST OF TABLES --- p.xvii === LIST OF FIGURES --- p.xix === Chapter Chapter 1 --- General Introduction === Chapter 1.1 --- Overview of lectins --- p.1 === Chapter 1.1.1 --- History of lectins --- p.1 === Chapter 1.1.2 --- Definitions of lectins --- p.2 === Chapter 1.1.3 --- Classification and nomenclature of lectins based on structure --- p.2 === Chapter 1.1.4 --- Classification and nomenclature of lectins based on carbohydrate-bindingspecificity --- p.4 === Chapter 1.1.5 --- Structure of plant lectins --- p.4 === Chapter 1.1.6 --- Biological function of plant lectins --- p.5 === Chapter 1.1.6.1 --- Anti-viral activity of plant lectiins --- p.5 === Chapter 1.1.6.2 --- Lectins as plant defense proteins --- p.6 === Chapter 1.1.6.3 --- Insecticidal activity of plant lectins --- p.7 === Chapter 1.1.6.4 --- Anti-fungal activity of plant lectins --- p.7 === Chapter 1.1.6.5 --- Mitogenic activity of plant lectins --- p.7 === Chapter 1.1.6.6 --- Anti-tumor and anti-proliferative activity of plant lectins --- p.9 === Chapter 1.1.7 --- Background of legume lectins --- p.11 === Chapter 1.1.7.1 --- Structure of legume lectins --- p.11 === Chapter 1.1.7.2 --- Functions and activities of legume lectins --- p.12 === Chapter 1.2 --- Overview of serine protease inhibitors in plants --- p.14 === Chapter 1.2.1 --- Classification of serine protease inhibitor --- p.15 === Chapter 1.2.2 --- The main functions of plant serine protease inhibitors --- p.17 === Chapter 1.2.3 --- Commercial application of serine protease inhibirtors --- p.19 === Chapter 1.2.3.1 --- Medical application --- p.19 === Chapter 1.2.3.2 --- Transgenic application in agriculture --- p.22 === Chapter 1.3 --- Overview of Pathogenesis-related proteins in plants --- p.25 === Chapter 1.3.1 --- Overview of PR-5 family Thaumatin-like proteins (TLPs) --- p.27 === Chapter 1.3.1.1 --- Structural similarities among TLPs --- p.28 === Chapter 1.3.1.2 --- Antifungal activity of TLP --- p.31 === Chapter 1.3.2 --- Overview of Chinase-like proteins (CLPs) --- p.33 === Chapter 1.3.2.1 --- Classification of chitinase --- p.34 === Chapter 1.3.2.1.1 --- On the basis of amino acid sequence of glycosyl hydrolase --- p.34 === Chapter 1.3.2.1.2 --- On the basis of amino acid sequence of plant chitinase --- p.35 === Chapter 1.3.2.2 --- Antifungal activity of CLP --- p.36 === Chapter 1.3.3 --- Anti-freeze property of PR proteins --- p.38 === Chapter 1.3.4 --- Application of PR proteins in agriculture --- p.40 === Chapter 1.4 --- Rationale of the present study --- p.42 === Chapter Chapter 2 --- Materials and Methods === Chapter 2.1 --- Materials --- p.43 === Chapter 2.2 --- Preparation of crude extract --- p.44 === Chapter 2.2.1 --- Hokkaido large black soybean --- p.44 === Chapter 2.2.2 --- Emperor banana --- p.45 === Chapter 2.3 --- Purification --- p.45 === Chapter 2.4 --- Chromatography --- p.46 === Chapter 2.4.1 --- DEAE-cellulose chromatography --- p.46 === Chapter 2.4.2 --- Affi-gel Blue gel --- p.47 === Chapter 2.4.3 --- SP-Sepharse --- p.48 === Chapter 2.4.4 --- Mono Q HR 5/5 and Mono S HR 5/5 --- p.49 === Chapter 2.4.5 --- Superdex 75 and superdex 200 --- p.50 === Chapter 2.5 --- Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) --- p.50 === Chapter 2.6 --- Protein concentration determination --- p.54 === Chapter 2.7 --- Preparation of rabbit reticulocyte lysate --- p.54 === Chapter 2.8 --- Determination of N-terminal amino acid sequence --- p.56 === Chapter 2.9 --- Assay of inhibition of hemagglutinating activity by different carbohydrates --- p.56 === Chapter 2.10 --- Thermal stability determination assays --- p.57 === Chapter 2.10.1 --- Stability at various temperatures --- p.57 === Chapter 2.10.2 --- Stability at 100°C --- p.57 === Chapter 2.11 --- Assay of pH dependence of hemagglutinating activity --- p.58 === Chapter 2.12 --- Assay of ion dependence of hemagglutinating activity --- p.58 === Chapter 2.13 --- Assay of antifungal activity --- p.58 === Chapter 2.14 --- Assay of trypsin inhibitory activity --- p.60 === Chapter 2.15 --- Assay of antibacterial activity --- p.61 === Chapter 2.16 --- Assay for cytotoxic activity on cancer cell lines --- p.61 === Chapter 2.17 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.62 === Chapter 2.18 --- Assay of mitogenic activity --- p.63 === Chapter Chapter 3 --- Purification and Characterization of Defense-Related Proteins from their Respective Sources === Chapter 3.1 --- Purification and Characterization of a Lectin from the Seeds of Hokkaido large black soybean === Chapter 3.1.1 --- Introduction --- p.65 === Chapter 3.1.2 --- Results --- p.66 === Chapter 3.1.3 --- Purification --- p.68 === Chapter 3.1.3.1 --- Affinity chromatography on Affi-gel Blue gel --- p.69 === Chapter 3.1.3.2 --- Anion-exchange chromatography on DEAE-cellulose --- p.70 === Chapter 3.1.3.3 --- Anion-exchange chromatography on Mono Q column --- p.71 === Chapter 3.1.3.4 --- Gel filtration on Superdex 200 column --- p.72 === Chapter 3.1.3.5 --- Hemagglutinating activity at each purification step --- p.73 === Chapter 3.1.4 --- Characterization of Lectin --- p.74 === Chapter 3.1.4.1 --- Molecular mass determination --- p.74 === Chapter 3.1.4.2 --- N-terminal amino acid sequencing --- p.76 === Chapter 3.1.4.3 --- Assay of inhibition of hemagglutinating activity by different carbohydrates --- p.77 === Chapter 3.1.4.4 --- Thermal stability --- p.78 === Chapter 3.1.4.5 --- Assay of pH dependence of hemagglutinating activity --- p.80 === Chapter 3.1.4.6 --- Assay of ion dependence of hemagglutinating activity --- p.81 === Chapter 3.1.4.7 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.82 === Chapter 3.1.4.8 --- Assay of mitogenic activity --- p.83 === Chapter 3.1.4.9 --- Assay of antibacterial activity --- p.84 === Chapter 3.1.5 --- Discussion --- p.86 === Chapter 3.2 --- Purification and Characterization of a Trypsin inhibitor from the Seeds of Hokkaido large black soybean === Chapter 3.2.1 --- Introduction --- p.93 === Chapter 3.2.2 --- Results --- p.94 === Chapter 3.2.3 --- Purification --- p.95 === Chapter 3.2.3.1 --- Anion-exchange chromatography on Mono Q column --- p.96 === Chapter 3.2.3.2 --- Gel filtration on Superdex 75 column --- p.98 === Chapter 3.2.3.3 --- Trypsin inhibitory activity at each purification step --- p.99 === Chapter 3.2.4 --- Characterization of trypsin inhibitory --- p.100 === Chapter 3.2.4.1 --- Molecular mass determination --- p.100 === Chapter 3.2.4.2 --- N-terminal amino acid sequencing --- p.102 === Chapter 3.2.4.3 --- Assay for HIV-1 reverse transcriptase (RT) inhibitory activity --- p.103 === Chapter 3.2.4.4 --- Antiproliferative effect on MCF-7 and Hep G2 cells --- p.104 === Chapter 3.2.4.5 --- pH and thermal stability --- p.105 === Chapter 3.2.5 --- Discussion --- p.106 === Chapter 3.3 --- Purification and Characterization of a Thaumatin-like protein and Chitinase-like protein from Emperor Banana === Chapter 3.3.1 --- Introduction --- p.108 === Chapter 3.3.2 --- Results --- p.109 === Chapter 3.3.3 --- Purification --- p.111 === Chapter 3.3.3.1 --- Affinity chromatography on Affi-gel Blue gel --- p.112 === Chapter 3.3.3.2 --- Cation exchange chromatography on Mono S column --- p.113 === Chapter 3.3.3.3 --- Gel filtration on Superdex 75 column --- p.114 === Chapter 3.3.3.3.1 --- Fraction MS 2 --- p.114 === Chapter 3.3.3.3.2 --- Fraction MS 4 --- p.115 === Chapter 3.3.3.3.3 --- Fraction MS 5 --- p.118 === Chapter 3.3.4 --- Characterization of the thaumatin-like protein --- p.121 === Chapter 3.3.4.1 --- N-terminal amino acid sequence determination --- p.121 === Chapter 3.3.4.2 --- Assay for antifungal activity --- p.122 === Chapter 3.3.4.3 --- Thermal stability --- p.124 === Chapter 3.3.4.4 --- pH stability --- p.125 === Chapter 3.3.4.5 --- Resistance to trypsin digestion --- p.125 === Chapter 3.3.4.6 --- Anti-HIV-1 reverse transcriptase activity --- p.126 === Chapter 3.3.4.7 --- Discussion --- p.127 === Chapter 3.3.5 --- Characterization of the two chitinase-like protein --- p.131 === Chapter 3.3.5.1 --- N-terminal amino acid sequence determination --- p.131 === Chapter 3.3.5.1.1 --- Emperor banana MS2 CLP --- p.131 === Chapter 3.3.5.1.2 --- Emperor banana MS4 CLP --- p.132 === Chapter 3.3.5.2 --- Assay for antifungal activity --- p.133 === Chapter 3.3.5.3 --- Discussion --- p.136 === Chapter Chapter 4 --- general discussion --- p.138 === References --- p.144