title: Enzyme-accelerated and structure-guided crystallization of calcium carbonate: role of the carbonic anhydrase in the homologous system.
abstract: The calcareous spicules from sponges, e.g. from Sycon raphanus, are composed of almost pure calcium carbonate. In order to elucidate the formation of those structural skeletal elements, the function of the enzyme carbonic anhydrase (CA), isolated from this species, during the in vitro calcium carbonate-based spicule formation, was investigated. It is shown that the recombinant sponge CA substantially accelerates calcium carbonate formation in the in vitro diffusion assay. A stoichiometric calculation revealed that the turnover rate of the sponge CA during the calcification process amounts to 25 CO2s(-1) × molecule CA(-1). During this enzymatically driven process, initially pat-like particles are formed that are subsequently transformed to rhomboid/rhombohedroid crystals with a dimension of ~50 μm. The CA-catalyzed particles are smaller than those which are formed in the absence of the enzyme. The Martens hardness of the particles formed is ~4 GPa, a value which had been determined for other biogenic calcites. This conclusion is corroborated by energy-dispersive X-ray spectroscopy, which revealed that the particles synthesized are composed predominantly of the elements calcium, oxygen and carbon. Surprising was the finding, obtained by light and scanning electron microscopy, that the newly formed calcitic crystals associate with the calcareous spicules from S. raphanus in a highly ordered manner; the calcitic crystals almost perfectly arrange in an array orientation along the two opposing planes of the spicules, leaving the other two plane arrays uncovered. It is concluded that the CA is a key enzyme controlling the calcium carbonate biomineralization process, which directs the newly formed particles to existing calcareous spicular structures. It is expected that with the given tools new bioinspired materials can be fabricated.
authors: ERC Advanced Investigator Grant Research Group at Institute for Physiological Chemistry, University Medical Center of the Johannes Gutenberg University Mainz, Duesbergweg 6, D-55128 Mainz, Germany. Electronic address: wmueller@uni-mainz.de.
authors: Werner E G
authors: WE
authors: Müller
authors: Ute
authors: U
authors: Schlossmacher
authors: Heinz C
authors: HC
authors: Schröder
authors: Ingo
authors: I
authors: Lieberwirth
authors: Gunnar
authors: G
authors: Glasser
authors: Michael
authors: M
authors: Korzhev
authors: Meik
authors: M
authors: Neufurth
authors: Xiaohong
authors: X
authors: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002119:Calcium Carbonate / Q000737:chemistry* / Q000378:metabolism*; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000378:metabolism*; D003460:Crystallization; D004602:Elements; D008903:Minerals / Q000737:chemistry; D008969:Molecular Sequence Data; D011161:Porifera / Q000033:anatomy & histology / Q000201:enzymology* / Q000648:ultrastructure; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D004602:Elements; D008903:Minerals; D002256:Carbonic Anhydrases; D002119:Calcium Carbonate
keywords: Calcareous spicules; Carbonic anhydrase; Crystal formation; Sponge; Sycon raphanus