A single crystal of charge-transfer 3,5-dimethylpyrazole benzilic acid (DMPBA) was grown at room temperature by using a slow evaporation solution growth technique. The grown crystal belonged to the monoclinic system with the space group of P21/n. Different spectroscopic and analytical techniques were used for analyzing the structure and properties of the grown crystal, such as Fourier transform infra-red, UV-Vis, 1H and 13C nuclear magnetic resonance. The mechanical strength of the crystal has been studied by using a Vickers’ micro-hardness test. The stiffness constant and yield strength of the crystal were also calculated from the micro-hardness test and Z-scan studies. The thermal stability of the crystal was studied by using thermo-gravimetric and differential thermal analysis and was found to be stable up to 187.6oC. In addition, the newly synthesized DMPBA compound was tested for deoxyribonucleic acid (DNA) binding, and in vitro-antimicrobial activity against various bacterial and fungal species. Also, the compound showed the moderate capacity of scavenging with 2,2-diphenyl-1-picrylhydrazyl (DPPH).