Lactocaseibacillus paracasei subsp. paracasei NTU 101 (NTU101) is a well-known commercial probiotic with multiple health beneficial effects. In this study, the gut microbiota modulation effect of an NTU 101 product, Vigiis 101-LAB, on healthy human was investigated in a randomized, double-blind, placebo-controlled human trial. Vigiis 101-LAB significantly modulated human gut microbiota at fourth and sixth weeks of trial (anosim analysis, P = 0.001). It also significantly improved peristalsis (P = 0.003) and shortened defecation interval of subjects. The shift of gut microbiota is significantly fit with defecation interval (P = 0.009) and stool shape (P = 0.001) of subjects. Our results suggest that Vigiis 101-LAB promotes human intestinal health with improvement of peristalsis and fecal quality. The gut modulation effects of Vigiis 101-LAB subsequently raised the abundance of vitamin B7, vitamin K, pyrimidine and purine biosynthesis pathways. Vigiis 101-LAB may promote peristalsis via purinergic pathway and possibly conferring prophylactic benefits against irritable bowel syndrome with constipation. © 2024 Society of Chemical Industry.