Biomineralization induced by microbial enzymes, which catalyse CaCO3 precipitation, is a promising field of research for various applications in building eco-materials. Especially, this could provide an eco-friendly process for protection of coastal areas against erosion. In the present investigation, fourteen bacterial strains were isolated and characterized from both natural seawater and calcareous deposits formed on a cathodically protected steel mesh in marine environment. All of them induced calcium carbonate precipitation in various media by producing urease and/or carbonic anhydrase enzymes. The calcium carbonate minerals produced by bacteria were identified by microscopy and µ-Raman spectroscopy. In parallel, an experimental set-up, based on a column reactor, was developed to study biomineralization and microbial capacity of Sporosarcina pasteurii to form sandy agglomerate. These well-known calcifying bacteria degraded the urea present in liquid medium circulating through the column to produce calcium carbonate, which acted as cement between sand particles. The bio-bricks obtained after 3 weeks had a compressive strength of 4.2 MPa. 20% of the inter-granular voids were filled by calcite and corresponded to 13% of the total mass. We successfully showed that bio-column system can be used to evaluate the bacterial ability to agglomerate a sandy matrix with CaCO3.
Marine bacterial biomineralisation by CaCO3 precipitation provides natural limestone structures, like beachrocks and stromatolites. Calcareous deposits can also be abiotically formed in seawater at the surface of steel grids under cathodic polarisation. In this work, we showed that this mineral-rich alkaline environment harbours bacteria belonging to different genera able to induce CaCO3 precipitation. We previously isolated 14 biocalcifying marine bacteria from electrochemically formed calcareous deposits and their immediate environment. By microscopy and µ-Raman spectroscopy, these bacterial strains were shown to produce calcite-type CaCO3. Identification by 16S rDNA sequencing provided between 98.5 and 100% identity with genera Pseudoalteromonas, Pseudidiomarina, Epibacterium, Virgibacillus, Planococcus, and Bhargavaea. All 14 strains produced carbonic anhydrase, and six were urease positive. Both proteins are major enzymes involved in the biocalcification process. However, this does not preclude that one or more other metabolisms could also be involved in the process. In the presence of urea, Virgibacillus halodenitrificans CD6 exhibited the most efficient precipitation of CaCO3. However, the urease pathway has the disadvantage of producing ammonia, a toxic molecule. We showed herein that different marine bacteria could induce CaCO3 precipitation without urea. These bacteria could then be used for eco-friendly applications, e.g., the formation of bio-cements to strengthen dikes and delay coastal erosion.
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