Caractérisation génétique et biochimique du système protéolytique de Streptococcus thermophilus : étude de la variabilité des systèmes de transport d’oligopeptides ; caractérisation des phénomènes d’ancrage, de maturation et de libération de la protéase PrtS ; production de peptides bioactifs à partir de caséines bovines

Abstract : We are interested in the production of bioactive peptides in fermented milk by the lactic acid bacterium Streptococcus thermophilus. For this, it requires that the bacterium internalize them as few as possible during its growth. Therefore, it was important to characterize the proteolytic system of S. thermophilus. First, phylogenetic relationships linking 30 S. thermophilus strains have been searched by MLST. Secondly, an ABC-type transport system which seems to be functional was identified in the LMD-9 strain and named OTS. A study of the variability of Ami and OTS transport systems of the 30 strains of S. thermophilus was performed. Finally, the hydrolysis of caseins by proteinase PrtS of S. thermophilus was studied. This proteinase usually anchored to the wall of the bacterium was also found in a free form in strain 4F44. The protein sequence of PrtS4F44, different from the one of PrtS in the LMD-9 strain (PrtSLMD-9), is not the cause of the partial release of PrtS4F44. Sortase A, the actor of the anchoring of PrtS to the wall of the bacteria, presents different alleles between the strain 4F44 (srtA4F44) and the LMD-9 strain (srtALMD-9). Indeed, PrtSLMD-9 is released when srtALMD-9 is replaced by srtA4F44 in the strain LMD-9 showing that SrtA4F44 is deficient, causing consequently a default of PrtS4F44 anchoring and its partial release into the extracellular medium. Additionally, hydrolysis of bovine caseinates was performed using the free form PrtS4F44 and allowed the production of bioactive peptides that can be used for the functionalization of fermented dairy products.
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Ahoefa Ablavi Awussi. Caractérisation génétique et biochimique du système protéolytique de Streptococcus thermophilus : étude de la variabilité des systèmes de transport d’oligopeptides ; caractérisation des phénomènes d’ancrage, de maturation et de libération de la protéase PrtS ; production de peptides bioactifs à partir de caséines bovines. Agronomie. Université de Lorraine, 2016. Français. ⟨NNT : 2016LORR0099⟩. ⟨tel-02131498⟩

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