To exploit the whole potential of Additive Manufacturing (AM), a sound knowledge about the mechanical and especially cyclic properties of AM materials as well as their dependency on the process parameters is indispensable. In the presented work, the influence of chemical composition of the used powder on the fatigue behavior of Selectively Laser Melted (SLM) and Laser Deposition Welded (LDW) specimens made of austenitic stainless steel AISI 316L was investigated. Therefore, in each manufacturing process two variations of chemical composition of the used powder were utilized. For qualitative characterization of the materials cyclic deformation behavior, load increase tests (LITs) were performed and further used for the physically based lifetime calculation method (PhyBaLLIT), enabling an efficient determination of stress (S)–number of cycles to failure (Nf) curves (S–Nf), which show excellent correlation to additionally performed constant amplitude tests (CATs). Moreover, instrumented cyclic indentation tests (PhyBaLCHT) were utilized to characterize the materials’ defect tolerance in a comparably short time. All material variants exhibit a high influence of microstructural defects on the fatigue properties. Consequently, for the SLM process a higher fatigue lifetime at lower stress amplitudes could be observed for the batch with a higher defect tolerance, resulting from a more pronounced deformation induced austenite–α’-martensite transformation. In correspondence to that, the batch of LDW material with an increased defect tolerance exhibit a higher fatigue strength. However, the differences in defect tolerance between the LDW batches is only slightly influenced by phase transformation and seems to be mainly governed by differences in hardening potential of the austenitic microstructure. Furthermore, a significantly higher fatigue strength could be observed for SLM material in relation to LDW specimens, because of a refined microstructure and smaller microstructural defects of SLM specimens.