Strain-Specific Ureolytic Microbial Calcium Carbonate Precipitation

Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, B-9000 Ghent, Belgium.
Applied and Environmental Microbiology (Impact Factor: 3.67). 09/2003; 69(8):4901-9. DOI: 10.1128/AEM.69.8.4901-4909.2003
Source: PubMed


During a study of ureolytic microbial calcium carbonate (CaCO3) precipitation by bacterial isolates collected from different environmental samples, morphological differences were observed
in the large CaCO3 crystal aggregates precipitated within bacterial colonies grown on agar. Based on these differences, 12 isolates were selected
for further study. We hypothesized that the striking differences in crystal morphology were the result of different microbial
species or, alternatively, differences in the functional attributes of the isolates selected. Sequencing of 16S rRNA genes
showed that all of the isolates were phylogenetically closely related to the Bacillus sphaericus group. Urease gene diversity among the isolates was examined by using a novel application of PCR-denaturing gradient gel
electrophoresis (DGGE). This approach revealed significant differences between the isolates. Moreover, for several isolates,
multiple bands appeared on the DGGE gels, suggesting the apparent presence of different urease genes in these isolates. The
substrate affinities (Km) and maximum hydrolysis rates (Vmax) of crude enzyme extracts differed considerably for the different strains. For certain isolates, the urease activity increased
up to 10-fold in the presence of 30 mM calcium, and apparently this contributed to the characteristic crystal formation by
these isolates. We show that strain-specific calcification occurred during ureolytic microbial carbonate precipitation. The
specificity was mainly due to differences in urease expression and the response to calcium.

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    • "The strain of B. lentus CP28 exhibited higher urease activity and more rapid growth and crystallization of calcium carbonate aggregation than strains B. diminuta CP16 and S. soli CP23. This observation coincides with the observations by Hammes et al. (2003), who reported a diversity of urease genes in the genomes of ureolytic bacteria and proposed that their high affinities and specific rates were the basase of rapid crystal formation. Urea is an organic nitrogenous compound present in coastal environments and introduced by the excretion of certain terrestrial and aquatic animals. "
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    ABSTRACT: Biomineralization is a known natural phenomenon associated with a wide range of bacterial species. Bacterial-induced calcium carbonate precipitation by marine isolates was investigated in this study. Three genera of ureolytic bacteria, Sporosarcina sp., Bacillus sp. and Brevundimonas sp. were observed to precipitate calcium carbonate minerals. Of these species, Sporosarcina sp. dominated the cultured isolates. B. lentus CP28 generated higher urease activity and facilitated more efficient precipitation of calcium carbonate at 3.24 ± 0.25 × 10(-4) mg/cell. X-ray diffraction indicated that the dominant calcium carbonate phase was calcite. Scanning electron microscopy showed that morphologies of the minerals were dominated by cubic, rhombic and polygonal plate-like crystals. The dynamic process of microbial calcium carbonate precipitation revealed that B. lentus CP28 precipitated calcite crystals through the enzymatic hydrolysis of urea, and that when ammonium ion concentrations reached 746 mM and the pH reached 9.6, that favored calcite precipitation at a higher level of 96 mg/L. The results of this research provide evidence that a variety of marine bacteria can induce calcium carbonate precipitation, and may influence the marine carbonate cycle in natural environments.
    06/2015; 46(2):455-64. DOI:10.1590/S1517-838246220140533
    • "This reaction is catalyzed by the enzyme urease, which is common in a wide range of soil microorganisms. It can be readily induced by adding inexpensive substrates and is involved in several biotechnological applications (Hammes et al. 2003; Stocks-Fischer, Galinat and Bang 1999). "
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    ABSTRACT: This paper describes a small-scale demonstration of an ex situ mixing technique that uses biocementation as an alternative to existing ground improvement techniques. The main objective of this research is to improve the strength and the stiffness of loose sand. Biocemented sand specimens have been produced by mixing a urease producing bacteria with nutrients and Sydney sand. Triaxial test with bender elements and the physical model test are conducted to investigate calcium carbonate precipitation and the properties of the soil. The model foundation tests have been performed at 1 g in a cylindrical tank with diameter of 600 mm. The tank has been filled with loose Sydney sand and a cemented column of 38 mm in diameter has been created in the center of the tank. A footing of 90 mm diameter has been placed on the sand surface and loaded to large displacements. The ability of the biocemented column made by ex situ mixing to significantly improve the foundation response is well demonstrated. Triaxial test perfo...
    05/2015; 9(1):48-56. DOI:10.1179/1937525515Y.0000000002
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    • "Furthermore, the proteins anchored on the cell wall could provide additional positive or negative sites for carbonate and calcium coordination. Calcite formed when Sporosarcina pasteurii (former Bacillus pasteurii) (Bang et al., 2001; Mitchell and Ferris, 2006) or other ureolytic bacteria were tested (Fujita et al., 2000) Similarly, B. sphaericus stabilized the calcite in solid media (Hammes et al., 2003) and when limestone was immersed in the liquid growth medium (Dick et al., 2006). On the contrary, when the nutrient broth and calcium acetate were substituted with calcium chloride, vaterite with a spherical morphology was identified for both B. sphaericus (De Muynck et al., 2008a) and S. pasteurii (Kim et al., 2013). "
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    ABSTRACT: In this work, Bacillus pumilus ACA-DC 4061 was selected for its increased capability for biomineralization on marble, under different nutrient media concentrations and temperature conditions. The optimum conditions for the CaCO3 bacterially-induced precipitation were determined with the aid of testing based on the Design of Experiments (DoE). Biomineral (vaterite) precipitation was favored in both the temperatures (25 and 30 °C) investigated. Stone loss rate was reduced when the samples were subjected to sonication. Thin sections of the substrate confirmed that vaterite was able to adhere onto the surface. Finally, under non-sterile conditions mimicking an in situ application, B. pumilus ACA-DC 4061 formed a fine layer of calcium carbonate. Therefore, this microorganism showed that vaterite formation may consistently occur under specific conditions and could prove useful as a candidate for on-site applications for stone conservation.
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