Transcription profiling-guided remodeling of sulfur metabolism in synthetic bacteria for efficiently capturing heavy metals

Highlights: • Transcription profiling reveals the role of sulfur metabolism in heavy metal capture. • A sulfur metabolism operon is de novo synthesized for constructing synthetic cells. • The synthesized operon remodels sulfur metabolism network in large scale. • The synthesized operon remarkably enhances heavy metal-tolerating capacity. • The synthesized operon remarkably promotes heavy metal capture in synthetic cells. Heavy metal contamination is becoming a global problem threatening human health. Heavy metal removal by engineered microbes by cellular adsorption and uptake is a promising strategy for treatment of heavy metal contamination. However, this strategy is confronted with limited heavy metal-capturing elements. In this study, we performed a transcription profiling-guided strategy for construction of heavy metal-capturing synthetic bacteria. Transcription profiling of a heavy metal-tolerating Cupriavidus taiwanensis strain revealed up-regulation of sulfur metabolism-related operons (e.g., iscSAU and moaEDAB) by Pb²⁺ and Cd²⁺. A synthetic Escherichia coli strain, EcSSMO, was constructed by design of a synthetic sulfur metabolism operon (SSMO) based on iscSAU/moaEDAB. Biochemical analysis and X-ray photoelectron spectroscopy (XPS) revealed that the synthetic bacteria had remodeled sulfur metabolism and enhanced heavy metal-tolerating capacity, with higher surviving EcSSMO cells than the surviving control cells Ec0 (not containing SSMO) at 50 mg/L of Pb²⁺ and Cd²⁺ (>92 % versus 90 % of Pb²⁺ and Cd²⁺ at 5 mg/L of Pb²⁺ and Cd²⁺, and >40 % of both heavy metals even at 50 mg/L of Pb²⁺ and Cd²⁺. This study reveals emphasizes feasibility of transcription profiling-guided construction of synthetic organisms by large-scale remodeling metabolic network.