Aim To investigate the relationship between single nucleotide polymorphism (SNP) of ATG16L1 gene promoter sequence and acute myocardial infarction (AMI). Methods The ATG16L1 gene promoter was amplified and sequenced by polymerase chain reaction in 285 AMI patients and 296 controls using case-control method. Combined with DNA sequencing sequence and alignment SNPs database, data statistics and analysis were carried out. After using the Hardy-Weinberg balance test, the χ² test and t test were used for correlation analysis. Logistic regression was used to analyze the association of multiple risk factors and three SNPs loci with susceptibility to AMI. Haploview 4.2 software and SHEsis online software were used for linkage disequilibrium and haplotype analysis. TRANSFAC database was used to predict the binding sites of transcription factors that may be affected by SNPs. Results Multivariate Logistic regression analysis showed that male, smoking history, and hypertension were independent risk factors for AMI (P＜0.05), while high density lipoprotein cholesterol was a protective factor for AMI (P＜0.05). Among the three SNPs (rs1816753, rs12476635, rs2289477) in the promoter sequence of ATG16L1 gene, the TC genotype of rs1816753 was associated with AMI, which significantly increased the risk of AMI (OR=2.9,5%CI:1.130～5.615, P＝0.024). Haploview 4.2 software analysis showed that the three SNPs were strongly linked. Conclusion The SNPs of ATG16L1 gene promoter may be associated with the susceptibility to AMI, and the TC genotype of rs1816753 may be a genetic risk factor for AMI.