Predicting Lactobacillus delbrueckii subsp. bulgaricus-Streptococcus thermophilus interactions based on a highly accurate semi-supervised learning method

Lactobacillus delbrueckii subsp. bulgaricus(L. bulgaricus) and Streptococcus thermophilus(S. thermophilus) are commonly used starters in milk fermentation. Fermentation experiments revealed that L. bulgaricus-S. thermophilus interactions(Lb St I) substantially impact dairy product quality and production. Traditional biological humidity experiments are time-consuming and labor-intensive in screening interaction combinations, an artificial intelligence-based method for screening interactive starter combinations is necessary. However,in the current research on artificial intelligence based interaction prediction in the field of bioinformatics, most successful models adopt supervised learning methods, and there is a lack of research on interaction prediction with only a small number of labeled samples.Hence, this study aimed to develop a semi-supervised learning framework for predicting Lb St I using genomic data from 362 isolates(181per species). The framework consisted of a two-part model: a co-clustering prediction model(based on the Kyoto Encyclopedia of Genes and Genomes(KEGG) dataset) and a Laplacian regularized least squares prediction model(based on K-mer analysis and gene composition of all isolates datasets). To enhance accuracy, we integrated the separate outcomes produced by each component of the two-part model to generate the ultimate Lb St I prediction results, which were verified through milk fermentation experiments. Validation through milk fermentation experiments confirmed a high precision rate of 85%(17/20; validated with 20 randomly selected combinations of expected interacting isolates). Our data suggest that the biosynthetic pathways of cysteine, riboflavin, teichoic acid, and exopolysaccharides, as well as the ATP-binding cassette transport systems, contribute to the mutualistic relationship between these starter bacteria during milk fermentation. However, this finding requires further experimental verification. The presented model and data are valuable resources for academics and industry professionals interested in screening dairy starter cultures and understanding their interactions.