Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis.

Microb Cell Fact

Department of Biology and Biochemistry, Milner Centre for Evolution, University of Bath, Claverton Down, Bath, BA2 7AY, UK.

Published: November 2021

AI Article Synopsis

  • Microbially induced calcite precipitation (MICP) is a bacterial process that has potential biotechnological applications, but is not fully understood due to unclear molecular mechanisms and a lack of insight into the contributing factors.
  • A study engineered Bacillus subtilis to induce MICP through the production of urease, which raises pH levels, and by enhancing this process with accessory genes for nutrient uptake and biofilm conditions.
  • The findings reveal key genetic elements and environmental interactions that influence MICP, providing a foundation for future engineering of bacteria for enhanced biomineralisation applications.

Article Abstract

Background: Microbially induced calcite precipitation (MICP) is an ancient property of bacteria, which has recently gained considerable attention for biotechnological applications. It occurs as a by-product of bacterial metabolism and involves a combination of chemical changes in the extracellular environment, e.g. pH increase, and presence of nucleation sites on the cell surface or extracellular substances produced by the bacteria. However, the molecular mechanisms underpinning MICP and the interplay between the contributing factors remain poorly understood, thus placing barriers to the full biotechnological and synthetic biology exploitation of bacterial biomineralisation.

Results: In this study, we adopted a bottom-up approach of systematically engineering Bacillus subtilis, which has no detectable intrinsic MICP activity, for biomineralisation. We showed that heterologous production of urease can induce MICP by local increases in extracellular pH, and this can be enhanced by co-expression of urease accessory genes for urea and nickel uptake, depending on environmental conditions. MICP can be strongly enhanced by biofilm-promoting conditions, which appeared to be mainly driven by production of exopolysaccharide, while the protein component of the biofilm matrix was dispensable. Attempts to modulate the cell surface charge of B. subtilis had surprisingly minor effects, and our results suggest this organism may intrinsically have a very negative cell surface, potentially predisposing it for MICP activity.

Conclusions: Our findings give insights into the molecular mechanisms driving MICP in an application-relevant chassis organism and the genetic elements that can be used to engineer de novo or enhanced biomineralisation. This study also highlights mutual influences between the genetic drivers and the chemical composition of the surrounding environment in determining the speed, spatial distribution and resulting mineral crystals of MICP. Taken together, these data pave the way for future rational design of synthetic precipitator strains optimised for specific applications.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8600894PMC
http://dx.doi.org/10.1186/s12934-021-01704-1DOI Listing

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