Abstract: In order to find the new targets to accelerate the solution of bacterial resistance problem caused by the antibiotic abuse, Escherichia coli BW25113 overexpressing acetyltransferase WcaB strain was constructed by using the pCA24N expression vector, and the killing effect of aminoglycoside antibiotics was tested in the logarithmic phase. Then, the minimum inhibitory concentration (MIC) and growth curve of Escherichia coli BW25113 overexpressing WcaB to aminoglycoside antibiotics were further determined. The results showed that: the overexpression of WcaB would inhibit the bacterial growth, and the MIC of aminoglycoside antibiotics to WcaB overexpressing strain did not change significantly. The overexpression of WcaB reduced the survival rate of bacteria treated with gentamicin or kanamycin, indicating that the overexpression of WcaB increased the sensitivity of Escherichia coli to aminoglycoside antibiotics. The results showed that acetyltransferase played an important role in enhancing the sensitivity of antibiotics to Escherichia coli, and the enzyme was expected to become a new pharmacological target for Escherichia coli.