The results showed that the treatment of constipation by L. rhamnosus Glory LG12 was associated with the regulation of Biosynthesis of cofactors, Biosynthesis of amino acids, Carbon metabolism, Ribosome, Amino sugar and nucleotide sugar metabolism, Pyrimidine metabolism, beta-N-acetylhexosaminidase activity, beta-galactosidase activity, alpha-L-fucosidase activity, beta- galactosidase activity, alpha-L-fucosidase activity, beta-galactosidase complex, NAD + kinase activity and other biological processes are closely related to metabolic pathways. This evidence concerns the gene GLB1 and constipation disorder.