© 2022, The Author(s), under exclusive licence to Springer Nature B.V.The number of studies on the removal of hazardous metals from water using genetic engineering technologies is growing. A high rate of metal ion removal from the environment is ensured, particularly through the expression of cysteine and thiol-rich proteins such as metallothioneins in bacterial cells. In this study, we used recombinant strains created by cloning the human metallothioneins MT2A and MT3 into Escherichia coli Jm109 to assess the removal and reduction of hexavalent chromium (Cr(VI)) from aqueous solutions. MT2A was the most effective strain in both Cr(VI) removal (89% in 25 mg/L Cr(VI)) and Cr(VI) reduction (76% in 25 mg/L Cr(VI)). The amount of Cr adsorbed per dry cell by the MT2A strain was 22 mg/g. The biosorption of total Cr was consistent with the Langmuir isotherm model. Scanning electron microscope (SEM) images revealed that the morphological structures of Cr(VI)-treated cells were significantly damaged when compared to control cells. Scanning transmission electron microscope (STEM) images showed black spots in the cytoplasm of cells treated with Cr(VI). Shifts in the Fourier transform infrared spectroscopy analysis (FTIR) spectra of the cells treated with Cr(VI) showed that the groups interacting with Cr were hydroxyl, amine, amide I, amide II, phosphoryl and carbonyl. When all of the experimental data was combined, it was determined that both MT2A and MT3 were effective in removing Cr(VI) from aqueous solutions, but MT2A was more effective, indicating that MT2A may be employed as a biotechnological tool. Graphical abstract: [Figure not available: see fulltext.].