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. 2025 May 28;15(1):18706.
doi: 10.1038/s41598-025-01350-2.

In situ mineral formation on a plate for direct and efficient screening of bacteria suitable for biocementation

Kazunori Nakashima  1 , Yuta Sugawara  2 , Masahiro Fujita  2 , Aya Yaegashi  2 , Meiqi Chen  2 , Sivakumar Gowthaman  3 , Thiloththama H K Nawarathna  3 , Wilson Mwandira  4 , Numbi Ramudu Kamini  5 , Parthasarathy Baskaran Sujiritha  5 , Chikara Takano  2 , Satoru Kawasaki  2
Affiliations

In situ mineral formation on a plate for direct and efficient screening of bacteria suitable for biocementation

Kazunori Nakashima et al. Sci Rep. .

Abstract

Microbially-induced carbonate precipitation (MICP) using a urea-hydrolyzing enzyme (urease) is a next-generation, environmentally friendly cementation technique. The isolation of excellent urease-producing bacteria for MICP is essential for establishing this technique. We have demonstrated a highly effective screening method for identifying promising bacteria suitable for MICP. In our selective plate, MICP-positive bacteria formed a CaCO3 halo around the colony on the plate, which was clearly distinguishable. Furthermore, we found that the CaCO3 formation activity of each bacterium was strongly correlated with the strength of the solidified sand samples. This technique is a novel approach for screening bacteria suitable for biocementation or MICP.

Keywords: Biocementation; Biomineralization; Carbonate precipitation; High-throughput screening; Selective plate.

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Conflict of interest statement

Declarations. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
(A) Possible factors on efficiency of MICP-based calcium carbonate precipitation by bacteria. (B) Strategy for direct and efficient screening of MICP-positive bacteria based on in situ mineral formation.
Fig. 2
Fig. 2
(A) Appearance of the colonies of Pararhodobacter sp. and E. coli inoculated on the selective plate M. (B) SEM-EDS analysis of white precipitate (halo) formed around the colony of Pararhodobacter sp.
Fig. 3
Fig. 3
Appearance of the colonies of bacteria inoculated on (A) NH4-YE medium-based plate (plate T-1) and (B) standard medium-based plate (plate T-2).
Fig. 4
Fig. 4
Urease activity of different bacteria species.
Fig. 5
Fig. 5
(A) CaCO3 formation test by the bacteria in a solution containing urea and CaCl2. (B) XRD analysis and SEM image of white precipitate formed in the CaCO3 formation test catalyzed by Pararhodobacter sp. Diamonds indicate the representative peaks of magnesium calcite.
Fig. 6
Fig. 6
Strength of solidified sand (top, middle, and bottom part of the specimen) by each bacterium.
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