The objective of this project is to demonstrate the lipolytic activity of probiotics in buttermilk fermentation with added milk phospholipids (MP), yielding products that can enhance MP absorption, providing additional health benefits through higher digestibility in intestine and possible modification of gut bacteria. Probiotics are beneficial for health, improving gut function, regulating immune response and the metabolites: lipases and proteases produced by them can hydrolysate lipids and proteins, which can lead to higher absorption of dairy nutrients. However, MP are known for its low absorption in intestine. Studies showed that metabolites produced by lactic acid bacteria (LAB) are associated with increased absorption of sphingomyelin and potentially all MP. Our hypothesis is that selected LAB would work better in helping digest MP. Bacteria isolation was performed from the LAB collection from OSU-Dr. Jimenez’s laboratory and further identified by 16S rDNA sequencing. Quantitative lipolysis activity was tested using α-naphthyl acetate. Eight promising strains were tested for probiotic potential by screening the ability to resist low pH and bile salts, high values in auto-aggregation and hydrophobicity, also must have antimicrobial activity, be susceptible to antibiotics and, screened their virulence factors by PCR. Three selected strains were grown in skim milk (as control) and compared with those grown in buttermilk with or without MP added. Lipolysis products were analyzed by TLC, HPLC, SDS-PAGE, zymograms and fatty acid sequencing after fermentation. The results showed that 7 of the promising strains with high lipolytic activity were probiotics. Three selected strains (Lactobacilluscasei, Lactobacillus helveticus, and Lactobacillus acidophilus) showed to grow better in buttermilk medium with added MP than other medium. They also have the enzymes to break down MP and produce lipolysis products. The results of this work can be used to increase bioavailability of MP and its application in fortified dairy products and pharmaceuticals.