Whole-cell bioreporter application for rapid evaluation of hazardous metal bioavailability and toxicity in bioprocess

Assessing heavy metal bioavailability and toxicity during bioprocess is critical for advancing green biotechnology. The capability of whole-cell bioreporters to measure heavy metal bioavailability has been increasingly recognized. The advantages of this technology being applied to bioprocess monitoring are less studied. Here we investigate the potential of a cadmium- and lead-sensitive bioreporter to be used for heavy metals as a class, which holds great interest for bioprocess applications. We evaluated the bioavailability of eight individual heavy metals with bioreporter zntA, as well as the bioavailability and toxicity of mixed metals. The bioavailability and toxicity of heavy metals in bioprocess samples were also evaluated. We have demonstrated for the first time that the zntA bioreporter can effectively detect the bioavailability of zinc, nickel, and cobalt with limit of detection lower than 0.01, 0.08 and 0.5 mg·L^-1, respectively. The detection limits meet the requirements of the WHO, the U.S. Environmental Protection Agency, and the China drinking water quality standards, which makes this approach reasonable for monitoring heavy metal bioavailability in bioprocess. LIVE/DEAD toxicity experiments have been conducted for the detection of mixed metal solution toxicity to zntA bioreporter which shows an EC₅₀ (as EC₅₀, concentration for 50% of maximal effect) value of mixed metal solution is 3.84 mg·L^-1. Samples from wastewater treatment plants, sludge treatment plants and kitchen waste fermentation processes were analyzed to extend upon the laboratory results. The results of this study confirm the potential for practical applications of bioreporter technology in bioprocess monitoring. In turn, development for such practical applications is key to achieve the necessary level of commercialization to further make the routine use of bioreporters in bioprocess monitoring feasible.