Ni(II) and Cu(II) complexes of Pyridoxal thiosemicarbazone (PLTSC) have been synthesized and characterized by ESI-MS, FTIR and UV-Vis spectral studies, elemental analyses, molar conductance, magnetic susceptibility measurements and thermal analysis. The spectral and analytical data suggest the tridentate nature of the ligand and formation of complexes in octahedral geometry. DNA-binding properties of these metal complexes with CT-DNA in a potassium phosphate buffer (pH 7.2) were investigated using UV-Vis absorption spectroscopy, fluorescence spectroscopy and viscosity measurements. Experimental studies suggest good DNA binding ability of the complexes. The cleavage of plasmid pBR322 DNA without any additives was evaluated using agarose gel electrophoresis and the complexes showed significant nuclease activity. The metal complexes are also screened for in vitro antibacterial activity and exhibited moderate activity against the tested organisms.