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 *292=:6292E>B

*292=:6292E>B
Avant tout, je remercie Dieu le tout puissant de m’avoir donner le courage et la patience pour
réaliser ce modeste travail.
Qu’il me soit permis d’exprimer ma reconnaissance à mon enseignant Mr. BELLAL M.M.,
Professeur à l’Institut National Agronomique d’El-Harrach, pour avoir accepté de diriger ce travail
malgré ses nombreuses occupations. C’est grâce à lui que je viens de faire mes premiers pas dans
la recherche.
Il m’est agréable d’exprimer mes sincères remerciements à mon co-promoteur, Mr. NOUANI
A., Maître assistant à l’université M’Hamed Bougara de Boumerdes, pour son aide précieuse, ses
encouragements et surtout pour l’accueil qu’il m’a toujours réservée dans son laboratoire. Je le
remercie également pour la confiance qu’il m’a témoignée. Ses qualités humaines et morales m’ont
aidé à mener à terme ce travail, qu’il veuille trouver ici mon estime, ma gratitude et mon respect.
Mon profond respect et mes sincères remerciements vont à mon enseignant, Mr.AMMOUCHE
A., Professeur à l’Institut National Agronomique d’El-Harrach, pour avoir accepter de présider le
jury et d’apprécier la qualité de ce travail.
Je tiens à remercier Mr. MATI A., Maître de conférence à l’université MOULOUD
MAMMERI de Tizi-Ouzou, pour avoir accepté de porter un jugement éclairé sur ce mémoire.
Mes remerciements vont à Mr. SABAOU N., Professeur à l’Ecole Normale Supérieure, qui
a bienvoulu examiner ce travail.
Je remercie vivement Melle. SLAMANI R., attachée de recherche à l’INRA, pour l’intérêt
qu’elle a manifesté dans ce travail. Je lui exprime ma gratitude pour la bienveillante attention
qu’elle m’a accordée tout au long de mon expérimentation. Mes sincères remerciements pour ses
conseils et pour sa grande compétence. Elle a su me donner du courage pour affronter les longues
journées passées au laboratoire. Je la remercie également pour tous ses sacrifices pour la réalisation
de ce mémoire.
Il existe une personne que je ne saurai jamais assez remercier ; c’est Mr. LABADI R.,
technicien au laboratoire de chimie des sols du département sciences du sol (INA), qui a
grandement contribué à l’aboutissement de ce mémoire. Il a su toujours me guider et me soutenir.
Je le remercie également pour sa gentillesse et sa sympathie. J’ai bénéficié de toutes ses qualités
humaines.
J’exprime ma reconnaissance à Melle. BENZITOUNI A., technicienne à l’Institut Pasteur
d’Algérie pour sa disponibilité à tout moment sans oublier Mme. MOULAHOUM A. pour ses
orientations précieuses. Je remercie également Melle. KABLI pour son aide.
Je m’en voudrais de ne pas remercier mon camarade MANSOURI A. pour toute la
documentation qu’il a mis à ma disposition et ce malgré son éloignement, sans oublier bien sur
mon amie Souhila pour tous ses encouragements.
Toute ma reconnaissance à Mabrouk et Larbi, techniciens au département de technologie
alimentaire (INA), pour leurs aides constantes, leurs conseils et surtout pour leur sympathie.
Mes remerciements vont également à notre bibliothécaire Baya pour sa disponibilité ainsi qu’à
tous mes amis qui, à diverses reprises, ont manifesté leurs soutien et leurs amitiés en particuliers
Rosa, Nora, Sabiha, Rachid, Nocia , Fayçal, Nabil et Madjid.
N
-F>=4:>6;ER%P?=6Z6:4>6;E%2>%:4=4:>D=6B4>6;E%@2%34%:;4N?34B2%@2%+?:;=%P?B633?B

*DB?9D
L’obtention d’une protéase coagulant le lait a été réalisée par culture en surface de Mucor pusillus.
L’extrait brut présente une force coagulante de l’ordre de 1 /1200.
Des essais de purification de l’extrait coagulant ont été réalisés. Le meilleur rendement en
activité a été obtenu par une chromatographie d’exclusion moléculaire sur Sephadex G-100 de
l’extrait brut lyophilisé.
L’électrophorèse sur PAGE-SDS a révélé une seule bande protéique homogène, caractérisée
par un poids moléculaire de l’ordre de 49 000 Da.
Les conditions optimales d’activité coagulante ont été déterminées. L’activité est maximale à
pH 5, à une température de 50°C et pour une concentration en CaCl2 du lait de 0,02 M. L’enzyme
est relativement stable dans l’intervalle de 30°C à 50°C pendant 30 mn d’incubation ainsi qu’aux
basses températures.
Ces propriétés sont relativement similaires à ceux de la présure traditionnelle, exception faite
pour la température.
Mots-clés : Protéases de Mucor pusillus , purification, caractérisation, coagulation du
lait, présure.

U
 &5B>=4:>

&5B>=4:>
The obtaining of the milk-clotting protease was carried out by solid-state culture of the strain of
Mucor pusillus. The crud extract show clotting strength about 1 / 1200.
The purification tests of the clotting extract were realised. A better recovery was obtained by
molecular size chromatography on Sephadex G-100 of the lyophilised crud extract.
The SDS- PAGE data was reveal a single homogeneous band, characterised by its molecular
weight measured about 49 000 Da.
The clotting optimal conditions activities were determined. The activity is maximum at a pH
5, at a temperature of 50°C and for a 0,02 M CaCl2 concentration. The enzyme is relatively stable
from 30°C at 50°C after 30 mn incubation period and at low temperature of conservation.
This properties were relatively similar with the rennet one except for the temperature.
Keys words: Mucor pusillus proteases, purification, characterisation, milk-clotting,
rennet.

V
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