Charbonnel Pascale, Lamarque Mauld, Piard Jean-Christophe, Gilbert Christophe, Juillard Vincent, Atlan Danièle
Useful Bacterial Surface Proteins, Unité de Recherches Laitières et Génétique Appliquée, Institut National de la Recherche Agronomique, 78352 Jouy-en-Josas Cedex, France.
J Biol Chem. 2003 Apr 25;278(17):14832-40. doi: 10.1074/jbc.M212454200. Epub 2003 Feb 16.
The specific oligopeptide transport system Opp is essential for growth of Lactococcus lactis in milk. We examined the biodiversity of oligopeptide transport specificity in the L. lactis species. Six strains were tested for (i) consumption of peptides during growth in a chemically defined medium and (ii) their ability to transport these peptides. Each strain demonstrated some specific preferences for peptide utilization, which matched the specificity of peptide transport. Sequencing of the binding protein OppA in some strains revealed minor differences at the amino acid level. The differences in specificity were used as a tool to unravel the role of the binding protein in transport specificity. The genes encoding OppA in four strains were cloned and expressed in L. lactis MG1363 deleted for its oppA gene. The substrate specificity of these engineered strains was found to be similar to that of the L. lactis MG1363 parental strain, whichever oppA gene was expressed. In situ binding experiments demonstrated the ability of OppA to interact with non-transported peptides. Taken together, these results provide evidence for a new concept. Despite that fact that OppA is essential for peptide transport, it is not the (main) determinant of peptide transport specificity in L. lactis.
特异性寡肽转运系统Opp对乳酸乳球菌在牛奶中的生长至关重要。我们研究了乳酸乳球菌中寡肽转运特异性的生物多样性。对六个菌株进行了以下测试:(i)在化学成分确定的培养基中生长期间对肽的消耗情况,以及(ii)它们转运这些肽的能力。每个菌株对肽的利用都表现出一些特定偏好,这与肽转运的特异性相匹配。对一些菌株中结合蛋白OppA进行测序,发现在氨基酸水平上存在微小差异。特异性的差异被用作揭示结合蛋白在转运特异性中作用的工具。克隆了四个菌株中编码OppA的基因,并在缺失其oppA基因的乳酸乳球菌MG1363中进行表达。发现无论表达哪个oppA基因,这些工程菌株的底物特异性都与乳酸乳球菌MG1363亲本菌株相似。原位结合实验证明了OppA与未转运肽相互作用的能力。综上所述,这些结果为一个新概念提供了证据。尽管OppA对肽转运至关重要,但它不是乳酸乳球菌中肽转运特异性的(主要)决定因素。