Synthesis of Calcium Carbonate Biological Materials: How Many Proteins are Needed?


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In Nature, calcium carbonate biomineralizations are the most abundant mineralized structures of biological origin. Because many exhibit remarkable characteristics, several attempts have been made to use them as substitution materials for bone reconstruction or as models for generating biomimetic composites that exhibit tailored properties. CaCO3 biomineralizations contain small amounts of amalgamate of proteins and polysaccharides that are secreted during the calcification process. They contribute to control the morphology of the crystallites and to spatially organize them in well-defined microstructures. These macromolecules, collectively defined as the skeletal matrix, have been the focus of a large number of studies aiming at synthesizing in vitro biomimetic materials, according to a bottom-up approach. However, recent proteomic investigations performed on the organic matrices associated to mollusc shells or to coral skeletons have quashed our hopes to generate, with only few macromolecular ingredients, biomimetic materials with properties approaching to those of natural biominerals. As a mean value, each matrix comprises a minimum of few tens of different proteins that seem to be strictly associated to calcium carbonate biominerals. Among the proteins that are currently detected, one finds RLCDs-containing proteins (Repetitive-Low-Complexity Domains), enzymes, proteins with protease inhibitors domains and at last, proteins that contains typical ECM (ExtraCellular Matrix) domains. Today, we still do not understand how the skeletal matrix works, and unveiling its complex functioning is one of the challenges for the coming decade, both from fundamental and applied viewpoints. Is it realistic to attempt generating abiotically, in a test tube at room temperature, biomimetic composites that mimic natural biomineralizations in their properties? If so, and by supposing that we know the individual functions of all the components of the matrix, is there a minimal number of proteins required for producing in vitro calcium carbonate biomaterials that approximate natural biominerals? These issues are of importance for the future research directions in biomaterials science.



Edited by:

Iulian Antoniac, Simona Cavalu and Teodor Traistaru




F. Marin et al., "Synthesis of Calcium Carbonate Biological Materials: How Many Proteins are Needed?", Key Engineering Materials, Vol. 614, pp. 52-61, 2014

Online since:

June 2014




* - Corresponding Author

[1] H.A. Lowenstam, S. Weiner, On Biomineralization, Oxford University Press, New-York, (1989).

[2] A.P. Jackson, J.F.V. Vincent, R.M. Turner, The mechanical design of nacre, Proc. R. Soc. Lond. B Biol. Sci. 234 (1988) 415-440.

[3] J. Aizenberg, A. Tkachenko, S. Weiner, L. Addadi, G. Hendler, Calcitic microlenses as part of the photoreceptor system in brittlestars, Nature, 412 (2001) 819-822.


[4] P. Westbroek, F. Marin, A marriage of bone and nacre, Nature 392 (1998) 861-862.

[5] S. Berland, O. Delattre, S. Borzeix, Y. Catonné, E. Lopez, Nacre/bone interface changes in durable nacre endosseous implants in sheep, Biomater. 26 (2005) 2767-2773.


[6] C. Tamerler, M. Sarikaya, Molecular biomimetics: utilizing nature's molecular ways in practical engineering. Acta Biomater. 3 (2007) 289-299.


[7] U. Ripamonti, J. Crooks, L. Khoali, L. Roden, The induction of bone formation by coral-derived calcium carbonate/hydroxyapatite constructs, Biomater. 30 (2009) 1428-1439.


[8] S. Mann, Biomineralization – Principles and Concepts in Bioinorganic Materials Chemistry, Oxford Chemistry Masters, (2001).

[9] J.W. Dunlop, P. Fratzl, Biological composites, Annu. Rev. Mater. Res. 40 (2010) 1-24.

[10] H. Nagasawa, M. Suzuki, Mechanism of nacre formation in mollusk shells : structure and function of organic matrices, in : S. Watabe, K. Maeyama, H. Nagasawa (Eds. ), Recent Advances in Pearl Research, Terrapub, Tokyo, 2013, pp.137-147.

[11] H. Nakahara, Nacre formation in bivalve and gastropod molluscs, in: S. Suga, H. Nakahara (Eds. ), Mechanisms and phylogeny of Mineralization in Biological Systems Springer-Verlag, Tokyo, 1991, pp.343-350.


[12] M. Rousseau, E. Lopez, P. Stempflé, M. Brendlé, L. Franke, A. Guette, R. Naslain, X. Bourrat, Multiscale structure of sheet nacre, Biomaterials 26 (2005) 6254-6262.


[13] D. Vilezeuf, N. Floquet, D. Chatain, F. Bonneté, D. Ferry, J. Garabou, E.M. Stolper, Multilevel modular mesocrystalline organization in red coral, Amer. Mineral. 95 (2010) 242-248.


[14] S. Weiner, Y. Levi-Kalisman, S. Raz, L. Addadi, Biologically formed amorphous calcium carbonate, Connect. Tissue Res. 44 (2003) 214-218.


[15] H. Cölfen, M. Antonietti, Mesocrystals: inorganic superstructures made by highly parallel crystallization and controlled alignment, Angew. Chem. Int. Ed. 44 (2005) 5576-5591.


[16] L.B. Gower, D.J. Odom, Deposition of calcium carbonate films by a polymer-induced liquid-precursor (PILP) process, J. Cryst. Growth 210 (2000) 719-734.


[17] Y.Y. Kim, L.B. Gower, Microcontact printing via a Polymer-Induced Liquid Precursor (PILP) process, Mat. Res. Soc. Symp. Proc. 724 (2002) 201-206.

[18] S.E. Wolf, I. Lieberwirth, F. Natalio, J.F. Bardeau, N. Delorme, F. Emmerling, R. Barrea, M. Kappl, F. Marin, Merging models of biomineralization with concepts of nonclassical crystallization : is a liquid amorphous precursor involved in the formation of the prismatic layer of the Mediterranean fan mussel Pinna nobilis ? Faraday Discussions 159 (2012).


[19] F. Marin, N. Le Roy, B. Marie, The formation and mineralization of mollusk shell, Front. Biosci. (Schol. Ed. ) 4 (2012) 1099-1125.

[20] B.T. Livingston, C.E. Killian, F. Wilt, A. Cameron, M.J. Landrum, O. Ermolaeva, V. Sapojnikov, D.R. Maglott, A.M. Buchanan, C.A. Ettensohn, A genome-wide analysis of biomineralization-related proteins in the sea urchin Strongylocentrotus purpuratus, Dev. Biol. 300 (2006).


[21] P. Westbroek, Life as a Geological Force – Dynamics of the Earth, W.W. Norton & Cie, New-York, (1991).

[22] S. Weiner, W. Traub, Macromolecules in mollusc shells and their function in biomineralization, Phil. Trans. R. Soc. Lond. B304 (1984) 425-434.

[23] Y. Levi-Kalisman, G. Falini, L. Addadi, S. Weiner, Structure of the nacreous organic matrix of a bivalve mollusc shell examined in the hydrated state using cryo-TEM, J. Struct. Biol. 135 (2001) 8-17.


[24] D.J. Jackson, C. McDougall, K. Green, F. Simpson, G. Wörheide, B.M. Degnan, A rapidly evolving secretome builds and patterns a sea shell. BMC Biology 4 (2006) 40-49.

[25] F. Marin, Molluscan shell matrix characterization by preparative SDS-PAGE, Scientif. World J. 3 (2003) 342-347.

[26] F. Marin, B. Marie, S. Benhamada, P. Silva, N. Le Roy, N. Guichard, S. Wolf, C. Montagnani, C. Joubert, D. Piquemal, D. Saulnier, Y. Gueguen, Shellome,: Proteins involved in mollusk shell biomineralization – diversity, functions, in: S. Watabe, K. Maeyama, H. Nagasawa (Eds. ), Recent Advances in Pearl Research, Terrapub, Tokyo, 2013, pp.149-166.

[27] F. Marin, G. Luquet, B. Marie, D. Medakovic, Molluscan shell proteins : primary structure, origin and evolution, Curr. Top. Dev. Biol. 80 (2008) 209-276.


[28] B. Marie, C. Joubert, A. Tayalé, I. Zanella-Cléon, C. Belliard, D. Piquemal, N. Cochennec-Loreau, F. Marin, Y. Gueguen, C. Montagnani, Different secretory repertoires control the biomineralization processes of prisms and nacre deposition of the pearl oyster shell, Proc. Natl. Acad. Sci. USA, 109 (2012).


[29] P. Ramos-Silva, J. Kaandorp, L. Huisman, B. Marie, I. Zanella-Cléon, N. Guichard, D.J. Miller, F. Marin, The skeletal proteome of the coral Acropora millepora: the evolution of calcification by co-option and domain shuffling. Mol. Biol. Evol. 30 (2013).


[30] B. Constantz, S. Weiner, Acidic macromolecules associated with the mineral phase of scleractinian coral skeletons, J. Exp. Zool. 248 (1988) 253-258.