We studied the characteristic X-ray spectra produced by the interaction of highly charged ions of 129Xeq+(q=25,26,27)with surface of metallic Mo. The experimental result shows that highly charged ions can excite the characteristic X-ray spectra of L-shell of Mo when the beam’s intensity is not more than 120 nA. The X-ray yield of single ion reaches a quantitative level of 10-8 and increases with the increment of the ion’s kinetic energy and ionic charge (potential energy). By measuring the X-ray spectra of Mo-Lα1, the M-level lifetime of Mo atom is estimated by using Heisenberg uncertainty relation.