Genetic Organization of the Operon Affects the Proteolytic Potential of Species in Milk.

Psychrotolerant species are a main cause of proteolytic spoilage of ultra-high temperature (UHT) milk products due to the secretion of the heat-resistant metallopeptidase AprX, which is encoded by the first gene of the operon. While the proteolytic property has been characterized for many different isolates, the underlying gene organization was only described for a few strains so far. In this study, the phylogenomic analysis of 185 type strains revealed that the presence of is strongly associated to a monophylum composed of 81 species, of which 83% carried the locus. Furthermore, almost all type strains of known milk-relevant species were shown to be members of the three monophyletic groups , , and . In total, 22 different types of genetic organizations were identified in the genus, whereby 31% of the species tested carried the type 1 operon structure consisting of eight genes (). Other genetic structures differed from type 1 mainly in the presence and location of genes coding for two lipases ( and ) and putative autotransporters ( and ). The peptidase activity of 129 strains, as determined on skim milk agar and in UHT-milk, correlated largely with different gene compositions. Particularly, isolates harboring the type 1 operon were highly proteolytic, while strains with other operon types, especially ones lacking and , exhibited significantly lower peptidase activities. In conclusion, the phylogenomic position and the gene organization specify the proteolytic potential of isolates. In addition, however, an interplay of several environmental factors and intrinsic traits influences production and activity of AprX, leading to strain-specific proteolytic phenotypes.