Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis

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 extrace...

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Autores principales:	Timothy D. Hoffmann, Kevin Paine, Susanne Gebhard
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Microbially induced calcite precipitation MICP Calcite Biomineralisation Urease Biofilm Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/6fcf476349f94a7fafed55bac6110190
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spelling	oai:doaj.org-article:6fcf476349f94a7fafed55bac61101902021-11-21T12:40:42ZGenetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis10.1186/s12934-021-01704-11475-2859https://doaj.org/article/6fcf476349f94a7fafed55bac61101902021-11-01T00:00:00Zhttps://doi.org/10.1186/s12934-021-01704-1https://doaj.org/toc/1475-2859Abstract 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.Timothy D. HoffmannKevin PaineSusanne GebhardBMCarticleMicrobially induced calcite precipitationMICPCalciteBiomineralisationUreaseBiofilmMicrobiologyQR1-502ENMicrobial Cell Factories, Vol 20, Iss 1, Pp 1-19 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbially induced calcite precipitation MICP Calcite Biomineralisation Urease Biofilm Microbiology QR1-502
spellingShingle	Microbially induced calcite precipitation MICP Calcite Biomineralisation Urease Biofilm Microbiology QR1-502 Timothy D. Hoffmann Kevin Paine Susanne Gebhard Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis
description	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.
format	article
author	Timothy D. Hoffmann Kevin Paine Susanne Gebhard
author_facet	Timothy D. Hoffmann Kevin Paine Susanne Gebhard
author_sort	Timothy D. Hoffmann
title	Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis
title_short	Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis
title_full	Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis
title_fullStr	Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis
title_full_unstemmed	Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis
title_sort	genetic optimisation of bacteria-induced calcite precipitation in bacillus subtilis
publisher	BMC
publishDate	2021
url	https://doaj.org/article/6fcf476349f94a7fafed55bac6110190
work_keys_str_mv	AT timothydhoffmann geneticoptimisationofbacteriainducedcalciteprecipitationinbacillussubtilis AT kevinpaine geneticoptimisationofbacteriainducedcalciteprecipitationinbacillussubtilis AT susannegebhard geneticoptimisationofbacteriainducedcalciteprecipitationinbacillussubtilis
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Genetic optimisation of bacteria-induced calcite precipitation in Bacillus subtilis

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