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| Pakistan Research Repository | Home |
Title of Thesis |
| Author(s) Mubashir Niaz |
| Institute/University/Department Details Botany University of Agriculture, Fasisalabad. |
| Session 2003 |
| Subject Botany |
| Number of Pages 154 |
| Keywords (Extracted from title, table of contents and abstract of thesis) |
Abstract |
| Download Full Thesis |  1339.2 KB |
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| S. No. | Chapter | Title of the Chapters | Page | Size (KB) |
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| 1 | 0 | Contents | 113.23 KB |
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| 2 | 1 | Introduction | 1 | 103.22 KB |
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| 3 | 2 | Review of literature | 13 | 83.59 KB |
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| 4 | 3 | Materials and methods | 22 | 213.32 KB |
| 3.1 | Microbial Strain | 22 | ||
| 3.1.1 | Slants Preparation | 22 | ||
| 3.1.2 | 1noculum preparation | 22 | ||
| 3.13 | Solid phase growth studies of arachniotus citrinus | 23 | ||
| 3.1.4 | Extraction of gluconamylasses | 23 | ||
| 3.1.5 | Protein Estimation | 24 | ||
| 3.1.6 | Glucoamylase assay | 25 | ||
| 3.1.7 | Calculations for glucoamyases activity units: | 26 | ||
| 3.2 | Purification of Glucoamylase | 27 | ||
| 3.2.1 | ammonium sulfate precipitation | 27 | ||
| 3.2.2 | Fast protein liquid chromatography( FPLC) | 28 | ||
| 3.2.3 | Hiload anioin -exchange chromatography | 28 | ||
| 3.2.4 | Hydrophobic-interaction chromatography | 29 | ||
| 3.2.5 | Mono-Q anio - exchange chromatography | 29 | ||
| 3.3 | Cargoxy1 group modification of glucoamylase | 29 | ||
| 3.3.1 | Native enzyme mobility shift assay (NEMSA) | 31 | ||
| 3.3.2 | Hirachin - 7% page | 31 | ||
| 3.3.3 | Native molecular weight | 32 | ||
| 3.4 | Carboxy group modificaito for alteration of characteristics | 38 | ||
| 3.4.1 | Temperature optimum | 39 | ||
| 3.4.2 | activation energy (Ea) | 39 | ||
| 3.5 | pH optimum | 39 | ||
| 3.6 | Effect of substrate( V max , K m , K eat ) | 40 | ||
| 3.6.1 | Thermodynamics of K eat | 40 | ||
| 3.6.2 | Thermodynamic of soluble starch hydrolysis | 42 | ||
| 3.7 | Irreversible thermal inactivation | 43 | ||
| 3.7.1 | activation energy of thermal denaturation | 44 | ||
| 3.7.2 | Thermodynamics of irreversible thermal inactivation | 44 | ||
| 3.8 | Determination of stability | 45 | ||
| 3.8.1 | Effect of Urea | 46 | ||
| 3.82 | Effect of Urea | 46 | ||
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| 5 | 4 | Results and discussions | 47 | 692.02 KB |
| 4.1 | Production of glucoamylase | 47 | ||
| 4.2 | Purification glucoamylase | 47 | ||
| 4.2.1 | Hiload ammonium sulfate precipitation | 48 | ||
| 4.2.2 | Hiload anion exchange chromatography | 49 | ||
| 4.2.3 | Hydrophobic Interaction chromatography | 51 | ||
| 4.2.4 | Mono-Q anoin exchange chromatography | 52 | ||
| 4.3 | Chemical modification of glucoamylase | 53 | ||
| 4.4 | Impact of carboxy1 group Modification on temperature optima | 61 | ||
| 4.5 | Effect of chemical modification on pH optima of glucoamylases | 68 | ||
| 4.6 | Thermohilicity: | 76 | ||
| 4.6.1 | Effect of additional aromatic interactions on kinetics of soluble starch hydrolysis | 76 | ||
| 4.6.2 | Effect of hydrophobization on kinetics of soluble starch hydrolysis | 81 | ||
| 4.7 | Effect of chemical modifications on thermostability of glucoamylase | 95 | ||
| 4.7.1 | Irreversible thermal stability | 96 | ||
| 4.8 | Stability to proteases | 114 | ||
| 4.9 | Stability to urea | 118 | ||
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| 6 | 5 | Summary | 124 | 692.02 KB |
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| 7 | 6 | Literature cited | 136 | 692.02 KB |
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| 8 | 7 | Appendices | 136 | 75.22 KB |
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