Intrinsic antimicrobial properties can give polyurethanes a wider range of applications. In this work, the bio-based chain extender, 2,5-diformylfuran dioxime (DFFD) possessing antimicrobial characteristic, is used to synthesize the intrinsically antimicrobial poly(oxime-urethane) (Dx-PU) with isophorone diisocyanate (IPDI) and poly(tetramethylene glycol) (PTMG-1000), and its properties are investigated. The results show that the tensile strength is 19.12 ± 0.49 MPa, the Young’s modulus of the poly(oxime-urethane) is 118.70 ± 11.40 MPa. The elongation at break and Shore A hardness were 292 ± 11 % and 93 ± 2, respectively. Additionally, the Dx-PU demonstrated a high thermal decomposition temperature of over 250 °C and a low glass transition temperature of the soft segments at approximately −60 °C. Meanwhile, research on Dx-PU coating has demonstrated that Dx-PU coating is 100 % antimicrobial against E. coli and S. aureus, and is suitable for use on a wide range of substrates, including glass, engineering plastics, metal and leather with excellent adhesion (ISO 0), pencil hardness (2H), impact resistance (0.5 m·kg) and flexibility (2 mm). The Dx-PU coating also has significant antibacterial performance in everyday environments. The outstanding flexibility, moderate Young’s modulus, high elongation at break, good thermal resistance, and excellent antimicrobial performance of Dx-PU make it highly suitable for applications on high-contact surfaces such as hospital walls, public sanitation facilities, countertops, and door handles. As a bio-based chain extender from renewable sources, DFFD offers new solutions for the synthesis of bio-based antimicrobial polyurethane and its coatings.