By adjusting the parameters in the additive manufacturing process, the optimization of the microstructure and mechanical properties of 2205 duplex stainless steel (DSS) is of great significance to ensure the reliability of the material in harsh environments. The 2205 DSS samples are prepared by cold metal transfer conditions of interlayer cooling time of 0, 30, 60, and 120 s, respectively, and the microstructure and properties of the samples are characterized in detail. With the increase of cooling time, the height of the samples increases, and the width decreases, and no defects are found in all the samples. It also reduces the grain size and contributes to the uniform distribution of grains. With the extension of cooling time, the proportion of austenitic phase gradually increases from 41.60% to 47.36%, while the proportion of ferrite decreases correspondingly. The mechanical properties of the samples are tested. With the increase of cooling time, the hardness of the samples increases from 646.32(±1.46) to 679.36(±7.27) MPa, and the ductility decreases from 60.10(±0.50)% to 43.67(±2.10)%. The results show that the interlayer cooling time can be used to regulate the formation, microstructure, and properties of DSS.