Detection of urease and carbonic anhydrase activity using a rapid and economical field test to assess microbially-induced carbonate precipitation

Abstract

Microbial precipitation of calcium carbonate has diverse engineering applications, from building and soil restoration, to carbon sequestration. Urease-mediated ureolysis and CO2 (de)hydration by carbonic anhydrase (CA) are known for their potential to precipitate carbonate minerals, yet many microbial community studies rely on marker gene or metagenomic approaches that are unable to determine in situ activity. Here, we developed fast and cost-effective tests for the field detection of urease and CA activity using pH-sensitive strips inside microcentrifuge tubes that change color in response to the reaction products of urease (NH3) and CA (CO2). Samples from a saline lake, a series of calcareous fens, and ferrous springs were assayed in the field, finding relatively high urease activity in lake samples, whereas CA activity was only detected in a ferrous spring. Incubations of lake microbes with urea resulted in significantly higher CaCO3 precipitation compared to incubations with a urease inhibitor. Therefore, the rapid assay indicated an on-site active metabolism potentially mediating carbonate mineralization. Field urease and CA activity assays complement molecular approaches and facilitate the search for carbonate-precipitating microbes and their in situ activity, which could be applied toward agriculture, engineering and carbon sequestration technologies.

High level of calcium carbonate precipitation achieved by mixed culture containing ureolytic and nonureolytic bacterial strains

“…Cellular urease can allow the strain to hydrolyse urea, whereas the presence of carbonic anhydrase facilitates rapid carbon transport into the cell via the CO 2 /HCO 3 interconversion pathway (Ferrer et al . 2020).…”

“…Interestingly, TBRC 5949 possesses both urease and carbonic anhydrase activities and the MICP process exhibited by this strain probably involves both urease and carbonic anhydrase enzymes. Cellular urease can allow the strain to hydrolyse urea, whereas the presence of carbonic anhydrase facilitates rapid carbon transport into the cell via the CO 2 / HCO 3 interconversion pathway (Ferrer et al 2020).…”