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Title of Thesis
PRIMARY METABOLITES OF CANDIDA TROPICALIS |
Author(s)
Atiya Abbasi |
Institute/University/Department Details
University of Karachi/ H.E.J Research Institute of Chemistry |
Session
1983 |
Subject
Chemistry |
Number of Pages
91 |
Keywords (Extracted from title, table of contents and abstract of thesis)
candida tropicalis, b-lactoglobulin , valinease, enzymic activity, enzymes, proteins, polysaccharides, lipids, synthetic peptides |
Abstract This is the first study to the characterization of proteins and peptides in the cell free broth of Candida tropicalis. We are reporting the presence of a highly specific proteolytic enzyme cleaving at valine. This has been confirmed by digesting B-lactoglobulin and a number of synthetic peptides. The enzyme has a carbohydrate moiety and a molecular weight of 40,000+7,000 as determined on SDS-plyacrylamide gel electrophoresis. Its optimum activity occurs at 37C at a ph between 8-9. We have named it Valinease because of its selective cleavage The peptides separated on Dowex 50 have been tested against a number of pathogenic and non-pathogenic bacteria. At least four of the peptides have been found to be active against most of the bacterial strains. The amino acid composition and N-terminal residues of these peptides have been determined
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| S. No. |
Chapter |
Title of the Chapters |
Page |
Size (KB) |
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| 1 |
0 |
Contents |
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 1914.81 KB |
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| 2 |
1 |
Introduction |
1 |
 11438.51 KB |
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1.1 |
General Background |
1 |
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1.2 |
Enzymes |
3 |
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1.3 |
Proteins |
7 |
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1.4 |
Polysaccharides |
11 |
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1.5 |
Lipids |
14 |
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1.6 |
Miscellanous |
15 |
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1.7 |
Biological Aspect |
19 |
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1.8 |
Pathogenesis by Ctropicalis |
21 |
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1.9 |
Experimentally induced Candidiasis in Animals |
24 |
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1.10 |
Objectives of present study |
25 |
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| 3 |
2 |
Experimental |
26 |
 11321.89 KB |
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2.1 |
Composition of Culture media |
26 |
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2.2 |
Preparation of culture media |
28 |
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2.3 |
Growth of the culture |
31 |
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2.4 |
Isolation of components from cell free broth |
31 |
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2.5 |
Gel filtration chromatography |
32 |
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2.6 |
Electgrophoresis |
33 |
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2.7 |
Affinity Chromatography |
36 |
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2.8 |
Enzyme assay |
37 |
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2.9 |
Oxidation with performic acid |
37 |
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2.10 |
Ion-Exchange chromatography |
38 |
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2.11 |
Amino acid analysis |
39 |
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2.12 |
Paper chromatography |
44 |
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2.13 |
Anti-Microbial assay |
46 |
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2.14 |
N-Terminal assay |
48 |
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2.15 |
Carbohydrate analysis |
49 |
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| 4 |
3 |
Results |
53 |
 8810.56 KB |
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3.1 |
Isolation of proteins and peptides |
53 |
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3.2 |
Electrophoresis of Proteins |
53 |
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3.3 |
Separation of proteins/peptides on sephadex |
57 |
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3.4 |
Purificatrion of enzyme |
57 |
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3.5 |
Electrophoresis of CTEpS1 |
59 |
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3.6 |
Amino acid composition of CTEpS1 |
59 |
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3.7 |
N-Terminal of CTEpS1 |
61 |
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3.8 |
Carbohydrate contents of CTEpS1 |
61 |
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3.9 |
Enzyme activity of CTEpS1 |
62 |
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3.10 |
Effect of ph on enzymic activity |
64 |
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3.11 |
Effect of temperature on enzymic activity |
64 |
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3.12 |
Splitting of B-lactoglogulin |
66 |
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3.13 |
Splitting of synthetic peptides |
69 |
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3.14 |
Separation of CT-Ep-F on dowex 50X2 |
71 |
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3.15 |
Amino acid analysis of peptides |
71 |
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3.16 |
N-Terminal assay of peptides |
73 |
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3.17 |
Antimicrobial activity of peptides |
73 |
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| 5 |
4 |
Discussion |
76 |
 8161.93 KB |
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4.1 |
Chemical Aspect |
76 |
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4.2 |
Biological aspect |
81 |
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4.3 |
References |
94 |
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