Obesity has been progressing at a rapid rate and possesses negative impacts on people’s well-being and health. In the present study, two glucagon-like peptide 1 (GLP-1) expression engineering strains, named as pMG36e-GLP-1 and pLIVE-GLP-1, were constructed, and their anti-obesity effects were evaluated in vivo. Our results indicated that both the pMG36e-GLP-1 and pLIVE-GLP-1 could produce GLP-1, and the mixture of these two strains could markedly reduce the blood glucose and weight of STZ-treated mice. When orally taking 109 CFU pMG36e-GLP-1 strains, the engineering strain could achieve a 14.73% weight loss of HFD-induced obesity mice, and could significantly reduce the blood glucose, weight, epididymis adipose mass, liver volume and lipid accumulation, as well as the TG, AST in mice serum and intracellular triglyceride in mice liver (p<0.05), and the western blot and q-PCR results indicated that pMG36e-GLP-1 and pMG36e-GLP-1+pLIVE-GLP-1 could significantly enhance the expression of genes involved in fatty acid oxidation, including ACOX1, ACSL, CPT1a and PPARa in liver tissues (p<0.05). In addition, the expression of GLUT4 was also increased after treated with pMG36e-GLP-1 and pMG36e-GLP-1+pLIVE-GLP-1. Our results provide evidence of the potential of pMG36e-GLP-1 used as a safe and effective non-absorbed oral treatment for obesity.