Disclosed are formulations for enhancing the in vivo colonization of probiotic microorganisms that include digestive enzymes and probiotic microorganisms, and polysorbate surfactants. The enzymes include lactogenic enzyme formulations that promote growth of Lactobacillus probiotics, bifidogenic enzyme tbrmulations that promote growth of Bifidobacterium probiotics and combination formulations that benefit both types of probiotics. It has been discovered. that certain polysorbate surfactants, including polysorbate-60 and polysorbate-80, further promote probiotic microorganism growth, when used with the enzyme formulations. The formulations are preferably compounded as dry powders, to avoid water reaction with the enzymes in blended formulations. Such formulations can be contained in capsules, tablets, packets or bottles and administered orally, either sequentially or in one combined formulation.