A comparative analysis of ball-milled biochar, graphene oxide, and multi-walled carbon nanotubes with respect to toxicity induction in Streptomyces

Ball-milled biochar has recently attracted a lot of attention due to the simplicity of its preparation and low cost. However, it is unknown if the biochar is environmentally safe. Here, the toxic effect of ball-milled biochar on Streptomyces was compared to that of pristine biochar and two other carbon nanomaterials of different shapes-graphene oxide and multi-walled carbon nanotubes. The effect of these different materials on antibiotic production was characterized. The results showed that even at concentrations of up to 10 mg/L, pristine biochar had a negligible effect on toxicity and antibiotic production in Streptomyces. However, after ball milling, the physical and chemical properties of biochar changed dramatically. Cells were severely damaged, and there was a significant increase in antibiotic production after the addition of ball-milled biochar. Exposure to 10 mg/L of ball-milled biochar caused massive cell disruption; the survival rate of Streptomyces coelicolor M145 cells was only 68.2% as compared to 90% after treatment with 10 mg/L graphene oxide and multi-walled carbon nanotubes. The secretion of the antibiotics- the red intracellular pigment undecylprodigiosin (RED) and blue diffusible pigment actinorhodin (ACT) was enhanced with the highest level in treatment with ball milled biochar, as compared to that with the other two carbon nanomaterials. This effect can be attributed to increased expression of pathway-specific regulatory genes redD, redZ and actil-ORF4. Ball-milled biochar can be developed as an effective additive to increase antibiotic yield. However, we should restrict the large-scale use of ball-milled biochar before fully understanding its impact on the environment and human health.