Pt-WOx-based catalyst has been considered as a promising candidate for the conversion of glycerol to 1,3-propanediol, but the catalytic performance was quite susceptible to the structure and chemical state of WOx species. Herein, we develop a facile approach to construct the single-atom tungsten-doped Al2O3 with a low chemical state through in-situ migration of WO3 nanoparticles under the H2 atmosphere. The characterization results demonstrated that the tetrahedral W5+ could disperse and stabilize the platinum species more effectively, and exhibited stronger interaction with platinum than the octahedral W6+ species. The enhanced catalytic performance is attributed to the rich oxygen defects on in-situ generated tetrahedral WOx on the catalyst, which facilitated the construction of Pt-WOx interface and strengthened the synergetic effects between them. The analysis of structure-performance correlation revealed that the mono-dispersed tetrahedral W5+ species [(-O-)3W(=O)] played a pivotal role in tuning catalyst acidity and improving the catalytic performance, which assists the adsorption of glycerol and captures H for in-situ formation of Brønsted acid site, and ultimately affected the selectivity toward 1,3-PDO and catalytic activity.