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coagulants of moringa oleifera lam. seeds purification ... - EPFL

coagulants of moringa oleifera lam. seeds purification ... - EPFL

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<strong>of</strong> Coagulating Activity <strong>of</strong> Different Compo-<br />

Measurement<br />

Present in M.<strong>oleifera</strong> Seed Extracts . . . . . . . . . . 91<br />

nents<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .92<br />

Abstract<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 92<br />

Introduction<br />

<strong>of</strong> Coagulation Efficiency . . . . . . . . . . . . . . . . . . . . . . . .92<br />

Testing<br />

<strong>of</strong> the Particle Concentration on<br />

Influence<br />

Coagulating Activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .94<br />

the<br />

<strong>of</strong> colloid material for coagulation experiments . . . . . . . . 95<br />

Choice<br />

<strong>of</strong> the Coagulation Rate . . . . . . . . . . . . . . . . . . . . . 96<br />

Measurement<br />

<strong>of</strong> Coagulation and Flocculation . . . . . . . . . . . . . . . . . 97<br />

Definition<br />

and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .99<br />

Materials<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .99<br />

Chemicals<br />

Seed Extracts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .99<br />

M.<strong>oleifera</strong><br />

Assay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .99<br />

Coagulation<br />

and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 101<br />

Results<br />

Extraction Procedures . . . . . . . . . . . . . . . . . . . . . . . . . 106<br />

Different<br />

<strong>of</strong> Fractions prepared by Chromatographic separation<br />

Analysis<br />

Extracts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108<br />

<strong>of</strong><br />

extract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 108<br />

Non-reduced<br />

after Reduction <strong>of</strong> the Sample . . . . . . . . . . . . . . . . . . 111<br />

Separation<br />

<strong>of</strong> Suspensions <strong>of</strong> Glassbeads and Bentonite . . . . . 114<br />

Flocculation<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 116<br />

Conclusions<br />

activity <strong>of</strong> M.<strong>oleifera</strong> extracts . . . . . . . . . . . . . . . . . 117<br />

Coagulating<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 120<br />

References<br />

<strong>of</strong> M.<strong>oleifera</strong> Seed Proteins . . . . . . 123<br />

Characterisation<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124<br />

Abstract<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 124<br />

Introduction<br />

Proteins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 126<br />

Seed<br />

and Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131<br />

Materials<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 131<br />

Chemicals<br />

extraction and quantification . . . . . . . . . . . . . . . . . . . . . 131<br />

Protein<br />

Separation (prep. HPLC + Äkta) . . . . . . . . . . 132<br />

Chromatographic<br />

SDS PAGE / Western Blotting . . . . . . . . . . . . . . . . 132<br />

Tris-Tricine<br />

Protein Staining and Immuno Staining . . . . . . . . . 132<br />

Non-Specific<br />

Staining <strong>of</strong> Glycosylated Proteins . . . . . . . . . . . . . . . . . 132<br />

Specific<br />

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 133<br />

MALDI/TOF-MS<br />

Mapping / Mass Mapping . . . . . . . . . . . . . . . . . . . . . . . 133<br />

Peptide<br />

and Discussion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 134<br />

Results<br />

Sequences <strong>of</strong> Selected Protein Fractions . . . . . . . . . . . . . . . 134<br />

Tag<br />

molecular weight proteins present in oil body fractions . . . 135<br />

Low<br />

Separated by Reverse Phase Chromatography<br />

Proteins<br />

Sample Reduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 137<br />

after<br />

digest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 139<br />

Tryptic<br />

Digest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 142<br />

Chymotryptic<br />

Immuno Detection <strong>of</strong> Proteins with Anti-Flo Antibodies . . . . . . 144<br />

v

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