Microbially induced calcium precipitation driven by denitrification: Performance, metabolites, and molecular...

Microbially induced calcium precipitation (MICP) driven by denitrification has attracted extensive attention due to its application potential in nitrate removal from calcium-rich groundwater. However, little research has been conducted on this technique at the molecular level. Here, Pseudomonas WZ39 was used to explore the molecular mechanisms of nitrate-dependent MICP and the effects of Ca²⁺ on bacterial transcriptional regulation and metabolic response. The results exhibited that appropriate Ca²⁺ concentration (4.5 mM) can promote denitrification and the production of ATP, EPSs, and SMPs. Genome-wide analysis showed that the nitrate-dependent MICP was accomplished through heterotrophic denitrification and CO₂ capture. During this process, EPS biosynthesis and Ca²⁺ signaling regulation were involved in the nucleation template supply and Ca²⁺ homeostasis balance. Untargeted transcriptome- and metabolome-association analyses revealed that the addition of Ca²⁺ triggered the significant up-regulation in several key pathways, such as transmembrane transporter and channel activities, amino acid metabolism, fatty acid biosynthesis, and carbon metabolism, which played a momentous role in the mineral nucleation and energy provision. The detailed information provided novel insights for understanding the active control of bacteria on MICP, and has great significance for deepening the cognition of groundwater remediation using nitrate-dependent MICP technique.