Centre for Molecular and Biomolecular Informatics, Radboud University Medical Centre, Nijmegen, the Netherlands.
BMC Genomics. 2010 Jan 15;11:36. doi: 10.1186/1471-2164-11-36.
Lactic acid bacteria (LAB) are a group of gram-positive, lactic acid producing Firmicutes. They have been extensively used in food fermentations, including the production of various dairy products. The proteolytic system of LAB converts proteins to peptides and then to amino acids, which is essential for bacterial growth and also contributes significantly to flavor compounds as end-products. Recent developments in high-throughput genome sequencing and comparative genomics hybridization arrays provide us with opportunities to explore the diversity of the proteolytic system in various LAB strains.
We performed a genome-wide comparative genomics analysis of proteolytic system components, including cell-wall bound proteinase, peptide transporters and peptidases, in 22 sequenced LAB strains. The peptidase families PepP/PepQ/PepM, PepD and PepI/PepR/PepL are described as examples of our in silico approach to refine the distinction of subfamilies with different enzymatic activities. Comparison of protein 3D structures of proline peptidases PepI/PepR/PepL and esterase A allowed identification of a conserved core structure, which was then used to improve phylogenetic analysis and functional annotation within this protein superfamily.The diversity of proteolytic system components in 39 Lactococcus lactis strains was explored using pangenome comparative genome hybridization analysis. Variations were observed in the proteinase PrtP and its maturation protein PrtM, in one of the Opp transport systems and in several peptidases between strains from different Lactococcus subspecies or from different origin.
The improved functional annotation of the proteolytic system components provides an excellent framework for future experimental validations of predicted enzymatic activities. The genome sequence data can be coupled to other "omics" data e.g. transcriptomics and metabolomics for prediction of proteolytic and flavor-forming potential of LAB strains. Such an integrated approach can be used to tune the strain selection process in food fermentations.
乳酸菌(LAB)是一群革兰氏阳性、产乳酸的厚壁菌。它们已被广泛应用于食品发酵,包括各种乳制品的生产。LAB 的蛋白水解系统将蛋白质转化为肽,然后转化为氨基酸,这对细菌生长至关重要,也是风味化合物作为终产物的重要贡献。高通量基因组测序和比较基因组杂交阵列的最新发展为我们提供了探索各种 LAB 菌株蛋白水解系统多样性的机会。
我们对 22 株已测序的 LAB 菌株的细胞壁结合蛋白酶、肽转运蛋白和肽酶等蛋白水解系统成分进行了全基因组比较基因组分析。PepP/PepQ/PepM、PepD 和 PepI/PepR/PepL 肽酶家族被描述为我们改进具有不同酶活性的亚家族区分的计算方法的示例。对脯氨酸肽酶 PepI/PepR/PepL 和酯酶 A 的蛋白质 3D 结构的比较,确定了一个保守的核心结构,然后用于改进该蛋白超家族的系统发育分析和功能注释。使用泛基因组比较基因组杂交分析探索了 39 株乳球菌(Lactococcus lactis)菌株的蛋白水解系统成分多样性。在不同乳球菌亚种或不同来源的菌株中,观察到蛋白酶 PrtP 及其成熟蛋白 PrtM、一种 Opp 转运系统和几种肽酶的变化。
改进后的蛋白水解系统成分的功能注释为未来预测的酶活性的实验验证提供了极好的框架。基因组序列数据可以与其他“组学”数据(例如转录组学和代谢组学)相结合,预测 LAB 菌株的蛋白水解和风味形成潜力。这种综合方法可用于调整食品发酵中的菌株选择过程。