In this study, sodium caseinate hydrolysates produced by papain with strong growth-stimulating activity for Streptococcus thermophilus (St) and Lactobacillus delbrueckii subsp. bulgaricus (Lb) were obtained. A series of separation methods including ultrafiltration, macroporous adsorption resin chromatography, gel filtration chromatography, and reverse-phase high-performance liquid chromatography (RP-HPLC) were applied to isolate and purify the peptide(s), which were mainly responsible for the activity. Finally, three novel growth-stimulating peptides, H-2-A, F2-c, and F2-b, corresponding to amino acid residues 29-35 and 103-108 of bovine α(S2)-casein and 181-186 of bovine α(S1)-casein, respectively, were obtained. With supplementation of H-2-A, F2-b, or F2-c at a protein concentration of 0.3%, the biomass yield of these two lactic acid bacteria (LAB) was enhanced by 193.3, 166.7, or 151.7%, respectively. In addition, there were significant (p < 0.05) increases in viable counts of St and lactic acid production of LAB in the presence of the purified peptides.
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