Sanchuan ham, produced in Yunnan, China, is food with ethnic characteristics favored by consumers. However, it can contain biogenic amines such as tyramine that are harmful to health, and the synthesis mechanism of biogenic amines in Sanchuan ham is not clear. This study focuses on the regulation of biogenic amine synthesis by quorum sensing. We used high‐performance liquid chromatography to detect the content of biogenic amine in different kinds of ham and found that the content of biogenic amine in Sanchuan ham was higher than that in others. Tyramine‐producing strain isolated from Sanchuan ham was identified as Enterococcus faecium. By monitoring the growth and tyramine synthesis of Enterococcus faecium under cultured conditions, the results found that high temperature and low salt increased tyramine production by E. faecium. After seven exogenous amino acids were applied to E. faecium, only tyrosine could promote the production of tyramine in E. faecium, and tyramine could not be synthesized in E. faecium until a certain amount was reached, indicating the presence of microbial quorum sensing signal molecules in the synthesis of tyramine in E. faecium. Untargeted metabolomics analysis of the differential metabolites produced by E. faecium showed that the contents of some peptides, especially alanyl‐leucine, were significantly increased. Further experiments with synthetic alanyl‐leucine illustrated that alanyl‐leucine activated the expression of tyrosine decarboxylase (tyrDC), thereby regulating the synthesis of tyramine by E. faecium. Alanyl‐leucine acted as quorum sensing signal molecules for biogenic amine synthesis by E. faecium, which provided a theoretical basis for reducing biogenic amine accumulation in ham. It is beneficial to control the content of biogenic amines in ham in the future.