Supplemental data for: Phenotypic differentiation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus isolates found in yogurt starter cultures

The mutualistic relationship between Streptococcus thermophilus (S. thermophilus) and L. del-brueckii subsp. bulgaricus (L. bulgaricus) is responsible for milk coagulation, gel formation, and the flavour of yogurt. Under set-style yogurt processing conditions, the performance of a mixed culture composed of these species depends on key technological parameters such as the capacity for acidi-fication and proteolytic activity. This study aimed to determine the extent of phenotypic diversity by comparing the key traits of acidification and proteolytic activity among isolates found in yo-gurt starter cultures. Seventy-two isolates from three industrial starter cultures were ranked by either their fast or slow acidification activity (time to reach pH 4.6, 16 h), proteolytic activity, cell envelope proteinase (CEP) activity, redox potential and titratable acidity. The integration of mul-tiple phenotype measures by hierarchical clustering and non-metric dimensional scaling (NMDS) clustered groups of isolates by multifactor similarity. A significant difference (p-value < 0.05) was observed between the clusters regarding redox potential and the proteolytic activity of both S. thermophilus and L. bulgaricus. The integration of multiple phenotypes points to the diversification that may have occurred over repeated culturing of yogurt starter bacteria. The phenotypic diver-sity may explain the divergence in starter performance and be used to refine the formulation of new starter cultures. Future work will investigate the correlation between the activity of specific enzymes based on the phenotype to explain the separation between the fast and slow acidification of isolates.