# Casein-assisted biomineralization of calcium carbonate microspheres for enhanced surface and adsorption properties

**Authors:** Aniket Gade, Julia Nadrowska, Joanna Trzcińska-Wencel, Marek Wiśniewski, Rajesh Raut, Mahendra Rai, Patrycja Golińska

PMC · DOI: 10.3389/fbioe.2025.1654712 · Frontiers in Bioengineering and Biotechnology · 2025-10-07

## TL;DR

This study shows that adding casein during the production of calcium carbonate microspheres improves their surface and adsorption properties, making them better for use as carrier materials.

## Contribution

The novel use of casein in biomineralization enhances microsphere properties for practical applications.

## Key findings

- Casein-assisted microspheres had a higher specific surface area (65 m²/g) compared to non-casein ones (47 m²/g).
- Casein improved morphology, pore volume, surface energy, and water adsorption capacity of the microspheres.
- Both microsphere types showed similar adsorption-desorption kinetics despite structural differences.

## Abstract

Biomineralization is a key biological process by which organisms form mineralized structures, with calcium carbonate being one of the most abundant naturally occurring biominerals. The development of synthetic analogs, particularly calcium carbonate microspheres (CaCO3-MS), holds potential for various applications, including as carrier materials.

In this study, CaCO3-MS were synthesized using a precipitation method, both with and without casein. Ammonium, sodium, and potassium carbonate were evaluated as precipitating agents to optimize microsphere formation. The physical properties of the resulting microspheres were characterized using nitrogen adsorption analysis, Brunauer-Emmett-Teller (BET) analysis, diffuse reflectance infrared Fourier transform spectroscopy (DRIFT), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray diffraction (XRD) analysis.

Ammonium carbonate was the most effective precipitating agent, yielding well-formed microspheres. Casein-assisted CaCO3-MS exhibited a higher specific surface area (65 m2/g) than CaCO3-MS synthesized without casein (47 m2/g). The casein-containing microspheres also demonstrated a more uniform spherical morphology, increased pore volume, higher surface energy, enhanced hydrophilicity, and approximately double the water adsorption capacity. However, both variants showed similar adsorption-desorption kinetics.

The presence of casein significantly improved the structural and functional properties of CaCO3-MS, making them more suitable for use as carrier materials. Furthermore, the described method enables the large-scale, surfactant-free synthesis of uniformly sized microspheres, enhancing its practical applicability.

## Linked entities

- **Proteins:** LOC105090951 (alpha-S2-casein)
- **Chemicals:** calcium carbonate (PubChem CID 10112), ammonium carbonate (PubChem CID 517111), sodium carbonate (PubChem CID 10340), potassium carbonate (PubChem CID 11430)

## Full-text entities

- **Chemicals:** Ammonium (MESH:D064751), Ammonium carbonate (MESH:C040502), calcium carbonate (MESH:D002119), water (MESH:D014867), nitrogen (MESH:D009584), potassium carbonate (MESH:C037593), sodium (MESH:D012964), CaCO3-MS (-)

## Full text

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## Figures

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## References

28 references — full list in the complete paper: https://tomesphere.com/paper/PMC12537877/full.md

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Source: https://tomesphere.com/paper/PMC12537877