Engineered Hydrophobin for Biomimetic Mineralization of Functional Calcium Carbonate Microparticles

Hanna Heinonen, Päivi Laaksonen, Markus B. Linder, Hans-Peter Hentze

VTT—Technical Research Centre of Finland, Espoo, Finland.

DOI: 10.4236/jbnb.2014.51001                

Abstract

In this study, the modified hydrophobin, engineered for biomimetic mineralization, has been employed as a structure-directing agent for mineralization of calcium carbonate. For the first time amphiphilic calcium carbonate particles have been obtained, using engineered proteins. The mineral microparticles have been characterized by optical microscopy, scanning electron microscopy (SEM) and X-ray diffraction (XRD). While mineralization in the presence of non-modified hydrophobin results in polymorph mineral structures, uniform microspheres with an average particle diameter of one micron are obtained by employing hydrophobin which has been modified with an additional ceramophilic protein sequence. Owing to the tri-functionality of the modified hydrophobin (hydrophilic, hydrophobic and ceramophilic), the obtained mineral microparticles exhibit amphiphilic properties. Potential applications are in the areas of functional fillers and pigments, like biomedical and composite materials. Pickering emulsions have been prepared as a demonstration of the emulsion-stabilizing properties of the obtained amphiphilic mineral microspheres. The structure-directing effects of the studied engineered hydrophobins are compared with those of synthetic polymers (i.e. polycarboxylates) used as crystallization and scaling inhibitors in industrial applications.

Keywords

Biomimetic Mineralization; Engineered Proteins; Hydrophobin; Calcium Carbonate; Functional Microparticles

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	H. Cölfen and S. Mann, “Higher-Order Organization by Mesoscale Self-Assembly and Transformation of Hybrid Nanostructures,” Angewandte Chemie International Edition, Vol. 42, No. 21, 2003, pp. 2350-2365. http://dx.doi.org/10.1002/anie.200200562
[2]	S. Mann, “Biomineralization—Principles and Concepts in Bioinorganic Materials Chemistry,” Oxford University Press, New York, 2001.
[3]	F. C. Meldrum, “Calcium Carbonate in Biomineralisation and Biomimetic Chemistry,” International Materials Reviews, Vol. 48, No. 3, 2003, pp. 187-224. http://dx.doi.org/10.1179/095066003225005836
[4]	M. Breulmann, H. Cölfen, H.-P. Hentze, M. Antonietti, D. Walsh and S. Mann, “Elastic Magnets: Template-Controlled Mineralization of Iron Oxide Colloids in a Sponge-Like Gel Matrix,” Advanced Materials, Vol. 10, No. 3, 1998, pp. 237-241. http://dx.doi.org/10.1002/(SICI)1521-4095(199802)10:3<237::AID-ADMA237>3.0.CO;2-6
[5]	P. Laaksonen, G. R. Szilvay and M. B. Linder, “Genetic Engineering in Biomimetic Composites,” Trends in Biotechnology, Vol. 30, No. 4, 2012, pp. 191-197. http://dx.doi.org/10.1016/j.tibtech.2012.01.001
[6]	A. Walther, I. Bjurhager, J.-M. Malho, J. Peere, J. Ruokolainen, L. A. Berglund and O. Ikkala, “Large-Area, Lightweight and Thick Biomimetic Composites with Superior Material Properties via Fast, Economic, and Green Pathways,” Nano Letters, Vol. 10, No. 8, 2010, pp. 2742-2748. http://dx.doi.org/10.1021/nl1003224
[7]	N. Yin, S. Y. Chen, Y. Ouyang, L. Tang, J. X. Jang and H.P. Wang, “Biomimetic Mineralization Synthesis of Hydroxyapatite Bacterial Cellulose Nanocomposites,” Progress in Natural Science: Materials International, Vol. 21, No. 6, 2011, pp. 472-477. http://dx.doi.org/10.1016/S1002-0071(12)60085-9
[8]	P. Fratzl, “Biomimetic Materials Research: What Can We Really Learn from Nature’s Structural Materials?” Journal of the Royal Society Interface, Vol. 4, No. 15, 2007, 637-642. http://dx.doi.org/10.1098/rsif.2007.0218
[9]	M. Li, H. Schnablegger and S. Mann, “Coupled Synthesis and Self-Assembly of Nanoparticles to Give Structures with Controlled Organization,” Nature, Vol. 402, 1999, pp. 393-395. http://dx.doi.org/10.1038/46509
[10]	H. Cölfen, “Precipitation of Carbonates: Recent Progress in Controlled Production of Complex Shapes,” Current Opinion in Colloid & Interface Science, Vol. 8, No. 1, 2003, pp. 23-31. http://dx.doi.org/10.1016/S1359-0294(03)00012-8
[11]	A. Peytcheva, H. Cölfen, H. Schnablegger and M. Antonietti, “Calcium Phosphate Colloids with Hierarchical Structure Controlled by Polyaspartates,” Colloid and Polymer Science, Vol. 280, No. 3, 2002, pp. 218-227. http://dx.doi.org/10.1007/s00396-001-0600-0
[12]	D. Gebauer and H. Cölfen, “Prenucleation Clusters and Non-Classical Nucleation,” Nano Today, Vol. 6, No. 6, 2011, pp. 564-584. http://dx.doi.org/10.1016/j.nantod.2011.10.005
[13]	H. A. B. Wosten, “Hydrophobins: Multipurpose Proteins,” Annual Review of Microbiology, Vol. 55, No. 1, 2001, pp. 625-646. http://dx.doi.org/10.1146/annurev.micro.55.1.625
[14]	M. B. Linder, G. R. Szilvay, T. Nakari-Setälä and M. E. Penttilä, “Hydrophobins: The Protein-Amphiphiles of Filamentous Fungi,” FEMS Microbiology Reviews, Vol. 29, No. 5, 2005, pp. 877-896. http://dx.doi.org/10.1016/j.femsre.2005.01.004
[15]	M. B. Linder, “Hydrophobins: Proteins That Self Assemble at Interfaces,” Current Opinion in Colloid & Interface Science, Vol. 14, No. 5, 2009, pp. 356-363. http://dx.doi.org/10.1016/j.cocis.2009.04.001
[16]	M. L. De Vocht, I. Reviakine, W. P. Ulrich, W. Bergsma-Schutter, H. A. B. Wosten, H. Vogel, A. Brisson, J. G. H. Wessels and G. T. Robillard, “Self-Assembly of the Hydrophobin SC3 Proceeds via Two Structural Intermediates,” Protein Science, Vol. 11, No. 5, 2002, pp. 1199-1205. http://dx.doi.org/10.1110/ps.4540102
[17]	G. R. Szilvay, A. Paananen, K. Laurikainen, E. Vuorimaa, H. Lemmetyinen, J. Peltonen and M. B. Linder, “Self-Assembled Hydrophobin Protein Films at the Air-Water Interface: Structural Analysis and Molecular Engineering,” Biochemistry, Vol. 46, No. 9, 2007, pp. 2345-2354. http://dx.doi.org/10.1021/bi602358h
[18]	P. Laaksonen, J. Kivioja, A. Paananen, M. Kainlauri, K. Kontturi, J. Ahopelto and M. B. Linder, “Selective Nanopatterning Using Citrate-Stabilized Au Nanoparticles and Cystein-Modified Amphiphilic Protein,” Langmuir, Vol. 25, No. 9, 2009, pp. 5185-5192. http://dx.doi.org/10.1021/la804001e
[19]	P. Laaksonen, M. Kainlauri, T. Laaksonen, A. Shchepetov, H. Jiang, J. Ahopelto and M. B. Linder, “Interfacial Engineering by Proteins: Exfoliation and Functionalization of Graphene by Hydrophobins,” Angewandte Chemie International Edition, Vol. 49, No. 29, 2010, pp. 4946-4949. http://dx.doi.org/10.1002/anie.201001806
[20]	M. B. Linder, M. Qiao, F. Laumen, K. Selber, T. Hyytia, T. Nakari-Setala, M. E. Penttila, “Efficient Purification of Recombinant Proteins Using Hydrophobins as Tags in Surfactant-Based Two-Phase Systems,” Biochemistry, Vol. 43, No. 37, 2004, pp. 11873-11882. http://dx.doi.org/10.1021/bi0488202
[21]	S. Varjonen, P. Laaksonen, A. Paananen, H. Valo, H. Hähl, T. Laaksonen and M. B. Linder, “Self-Assembly of Cellulose Nanofibrils by Genetically Engineered Fusion Proteins,” Soft Matter, Vol. 7, No. 6, 2011, pp. 2402-2411. http://dx.doi.org/10.1039/c0sm01114b
[22]	S. S. Behrens, “Synthesis of Inorganic Nanomaterials Mediated by Protein Assemblies,” Journal of Materials Chemistry, Vol. 18, 2008, pp. 3788-3798. http://dx.doi.org/10.1039/b806551a
[23]	D. Santhiya, Z. Burghard, C. Greiner, L. P. H. Jeurgens, T. Subkowski and J. Bill, “Bioinspired Deposition of TiO2 Thin Films Induced by Hydrophobins,” Langmuir, Vol. 26, No. 9, 2010, pp. 6494-6502. http://dx.doi.org/10.1021/la9039557
[24]	A. Schulz, B. M. Liebeck, D. John, A. Heiss, T. Subkowski and A. Boker, “Protein-Mineral Hybrid Capsules from Emulsions Stabilized with an Amphiphilic Protein,” Journal of Materials Chemistry, Vol. 21, 2011, pp. 9731-9736. http://dx.doi.org/10.1039/c1jm10662g
[25]	J. Rieger, E. Hädicke, I. U. Rau and D. Boeckh, “A Rational Approach to the Mechanism of Incrustation Inhibition by Polymeric Additives,” Tenside, Surfactants, Detergents, Vol. 34, 1997, pp. 430-435.
[26]	M. B. Linder, K. Selber, T. Nakari-Setälä, M. Qiao, M.-R. Kula and M. Penttilä, “The Hydrophobins HFBI and HFBII from Trichoderma reesei Showing Efficient Interactions with Nonionic Surfactants in Aqueous Two-Phase Systems,” Biomacromolecules, Vol. 2, No. 2, 2001, pp. 511-517. http://dx.doi.org/10.1021/bm0001493
[27]	K. Zhang, M. R. Diehl and D. A. Tirrell, “Artificial Polypeptide Scaffold for Protein Immobilization,” Journal of the American Chemical Society, Vol. 127, No. 29, 2005, pp. 10136-10137. http://dx.doi.org/10.1021/ja051457h
[28]	D. V. Volodkin, R. von Klitzing and H. Moehwald, “Pure Protein Microspheres by Calcium Carbonate Templating,” Angewandte Chemie International Edition, Vol. 49, No. 48, 2010, pp. 9258-9261. http://dx.doi.org/10.1002/anie.201005089
[29]	H. Imai, Y. Oaki and A. Kotachi, “A Biomimetic Approach for Hierarchically Structured Inorganic Crystals through Self-Organization,” Bulletin of the Chemical Society of Japan, Vol. 79, No. 12, 2006, pp. 1834-1851. http://dx.doi.org/10.1246/bcsj.79.1834
[30]	N. A. J. M. Sommerdijk and G. D. With, “Biomimetic CaCO3 Mineralization Using Designer Molecules and Interfaces,” Chemical Reviews, Vol. 108, No. 11, 2008, pp. 4499-4550. http://dx.doi.org/10.1021/cr078259o
[31]	M. Sedlák, M. Antonietti and H. Cölfen, “Synthesis of a New Class of Double-Hydrophilic Block Copolymers with Calcium Binding Capacity as Builders and for Biomimetic Structure Control of Minerals,” Macromolecular Chemistry and Physics, Vol. 199, No. 2, 1998, pp. 247-254. http://dx.doi.org/10.1002/(SICI)1521-3935(19980201)199:2<247::AID-MACP247>3.0.CO;2-9
[32]	Z.-G. Cui, C.-F. Cui, Y. Zhu and B. P. Binks, “Multiple Phase Inversion of Emulsions Stabilized by in Situ Surface Activation of CaCO3 Nanoparticles via Adsorption of Fatty Acids,” Langmuir, Vol. 28, No. 1, 2012, pp. 314-320. http://dx.doi.org/10.1021/la204021v
[33]	R. Aveyard, B. P. Binks and J. H. Clint, “Emulsions Stabilised Solely by Colloidal Particles,” Advances in Colloid and Interface Science, Vol. 100-102, 2003, pp. 503-546. http://dx.doi.org/10.1016/S0001-8686(02)00069-6
[34]	G. Jutz and A. Böker, “Bionanoparticles as Functional Macromolecular Building Blocks—A New Class of Nanomaterials,” Polymer, Vol. 52, No. 2, 2011, pp. 211-232. http://dx.doi.org/10.1016/j.polymer.2010.11.047
[35]	G. Falini, S. Albeck, S. Weiner and L. Addadi, “Control of Aragonite or Calcite Polymorphism by Mollusk Shell Macromolecules,” Science, Vol. 271, No. 5245, 1996, pp. 67-69. http://dx.doi.org/10.1126/science.271.5245.67
[36]	S. Weiner and L. Addadi, “Crystallization Pathways in Biomineralization,” Annual Review of Materials Research, Vol. 21, 2011, pp. 21-40. http://dx.doi.org/10.1146/annurev-matsci-062910-095803