PMID- 34794448
OWN - NLM
STAT- MEDLINE
DCOM- 20220131
LR  - 20220131
IS  - 1475-2859 (Electronic)
IS  - 1475-2859 (Linking)
VI  - 20
IP  - 1
DP  - 2021 Nov 18
TI  - Genetic optimisation of bacteria-induced calcite precipitation in Bacillus 
      subtilis.
PG  - 214
LID - 10.1186/s12934-021-01704-1 [doi]
LID - 214
AB  - 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.
CI  - (c) 2021. The Author(s).
FAU - Hoffmann, Timothy D
AU  - Hoffmann TD
AD  - Department of Biology and Biochemistry, Milner Centre for Evolution, University 
      of Bath, Claverton Down, Bath, BA2 7AY, UK.
FAU - Paine, Kevin
AU  - Paine K
AD  - Department of Architecture and Civil Engineering, BRE Centre for Innovative 
      Construction Materials, University of Bath, Bath, BA2 7AY, United Kingdom.
FAU - Gebhard, Susanne
AU  - Gebhard S
AUID- ORCID: 0000-0003-4783-6115
AD  - Department of Biology and Biochemistry, Milner Centre for Evolution, University 
      of Bath, Claverton Down, Bath, BA2 7AY, UK. s.gebhard@bath.ac.uk.
LA  - eng
GR  - EP/PO2081X/1/Engineering and Physical Sciences Research Council/
PT  - Journal Article
DEP - 20211118
PL  - England
TA  - Microb Cell Fact
JT  - Microbial cell factories
JID - 101139812
RN  - 0 (DNA, Bacterial)
RN  - 0 (Polysaccharides, Bacterial)
RN  - 0 (exopolysaccharide, Bacillus)
RN  - 7OV03QG267 (Nickel)
RN  - 8W8T17847W (Urea)
RN  - EC 3.5.1.5 (Urease)
RN  - H0G9379FGK (Calcium Carbonate)
SB  - IM
MH  - Bacillus subtilis/*genetics/*metabolism
MH  - Biofilms
MH  - *Biomineralization
MH  - Calcium Carbonate/chemistry/*metabolism
MH  - Chemical Precipitation
MH  - DNA, Bacterial
MH  - Genetic Engineering
MH  - Hydrogen-Ion Concentration
MH  - Industrial Microbiology/methods
MH  - Nickel/metabolism
MH  - Polysaccharides, Bacterial/*metabolism
MH  - Sequence Deletion
MH  - Urea/metabolism
MH  - Urease/*metabolism
PMC - PMC8600894
OTO - NOTNLM
OT  - Biofilm
OT  - Biomineralisation
OT  - Calcite
OT  - Genetic engineering
OT  - MICP
OT  - Microbially induced calcite precipitation
OT  - Synthetic biology
OT  - Urease
COIS- The authors declare that they have no competing interests.
EDAT- 2021/11/20 06:00
MHDA- 2022/02/01 06:00
PMCR- 2021/11/18
CRDT- 2021/11/19 05:36
PHST- 2021/08/18 00:00 [received]
PHST- 2021/11/06 00:00 [accepted]
PHST- 2021/11/19 05:36 [entrez]
PHST- 2021/11/20 06:00 [pubmed]
PHST- 2022/02/01 06:00 [medline]
PHST- 2021/11/18 00:00 [pmc-release]
AID - 10.1186/s12934-021-01704-1 [pii]
AID - 1704 [pii]
AID - 10.1186/s12934-021-01704-1 [doi]
PST - epublish
SO  - Microb Cell Fact. 2021 Nov 18;20(1):214. doi: 10.1186/s12934-021-01704-1.