BackgroundFolic acid is a class of B vitamins that have the function of improving intestinal microbiota.ResultLactiplantibacillus plantarum(L. plantarum) LZ227, which is a highly folate‐producing strain, was used as the research object, and the folic acid produced by LZ227 was further identified by LC‐MS, and the structural diversity, community composition, abundance difference, and short‐chain fatty acids(SCFAs) content in fermentation broth were studied by the manure slurry fermentation model. The results showed that the folic acid produced by LZ227 was 5‐methyltetrahydrofolate (5‐MTHF).ConclusionLZ227 can increase the intestinal microbial diversity in the state of folate‐free, regulate the intestinal flora, increase the abundance of Firmicutes in the intestinal flora, and inhibit the abundance of Bacteroidetes.LZ227 can inhibit the growth of Alistipes, Parabacteroides, and Bacteroides in the intestine. LZ227 significantly reduced the acetic acid content and significantly increased the butyric acid content in the case of folate free.This article is protected by copyright. All rights reserved.