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Aim: Nano-biotechnologically synthesizing silver nanoparticles via canthaxanthin pigment extracted from Dietzia maris AURCCBT01 and assessing their cytotoxic therapeutic potential against human keratinocyte cell line (HaCaT) were the key objectives of this study., Methods and Results: The pigment extracted from D. maris AURCCBT01 was identified as canthaxanthin using UV-VIS spectroscopy, FTIR, NMR ( 1 H NMR and 13 C NMR) and MS. Canthaxanthin, treated with silver nitrate solution, produced canthaxanthin-mediated silver nanoparticles and they were characterized by UV-VIS spectroscopy, FTIR, XRD, FESEM-EDX and TEM-SAED techniques. UV-VIS spectroscopy pointed out an absorption band at 420 nm, relating to the surface plasmon resonance of silver nanoparticles. FTIR findings suggested that the diverse functional groups of canthaxanthin bio-molecules played a significant task in capping the silver nanoparticles. XRD analysis exhibited 40·20 nm for the crystal size of nanoparticles. FESEM and TEM exhibited that the biosynthesized silver nanoparticles were spherical in shape with crystalline nature and the particle size was 40-50 nm. Moreover, the cytotoxicity assessment of the synthesized nanoparticles in HaCaT revealed significant cytotoxicity in the cultured cells with an IC 50 value of 43 µg ml -1 ., Conclusion: Stable silver nanoparticles synthesized using canthaxanthin from D. maris AURCCBT01 were found effective for application in wound healing activity., Significance and Impact of the Study: Biosynthesized silver nanoparticles via canthaxanthin bacterial pigment exhibited their cytotoxicity effect in HaCaT and testified their eventual therapeutic potential in the wound healing activity with no side effects in a cost effective and eco-friendly process., (© 2020 The Society for Applied Microbiology.)