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Application of Hypothetical Cathepsin-like Protein from Nematostella vectensis and Its Mutant Silicatein-like Cathepsin for Biosilica Production

Published online by Cambridge University Press:  28 August 2013

Mi-Ran Ki ,
Ki Ha Min  and
Seung Pil Pack
Mi-Ran Ki
Affiliation:
Department of Biotechnology and Bioinformatics, Korea University, 2511 Sejong-Ro, Sejong city 339-700, Korea
Ki Ha Min
Affiliation:
Department of Biotechnology and Bioinformatics, Korea University, 2511 Sejong-Ro, Sejong city 339-700, Korea
Seung Pil Pack
Affiliation:
Department of Biotechnology and Bioinformatics, Korea University, 2511 Sejong-Ro, Sejong city 339-700, Korea
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Abstract

Silicatein is general catalyst for synthesis of silica structure in siliceous sponges. However, the advent of biomimetic silicification by this recombinant version is limited by its poor yield. To overcome this limitation, we employed a cathepsin L as an alternative to silicatein. Cathepsin L has high sequence identity and similarity with silicatein alpha except cysteine other than serine residues at the active site. Here, we expressed recombinant hypothetical cathepsin-like protein (CAT) from Nematostella vectensis, displaying not only protease activity but also silica condensing activity. To increase the silica forming activity, some residues including cysteine in active site were changed into silicatein conserved residues. The mutant silicatein-like cathepsin (SLC) revealed increased protein stability in comparison with that of CAT when expressed in E. coli. The silica forming activity of SLC was comparable to that of SIL. SLC produced silica particles of size less than 50 nm which were increased to 200∼300 nm in the presence of a structure-directing agent, Triton X-100. Protein immobilization by SLC-mediated silicification was performed using bovine carbonic anhydrase under ambient conditions. Immobilized protein retained its enzymatic activity for a longer time and was reused up to several times. In conclusion, CAT from Nematostella vectensis was evolved to a more soluble and available biosilica forming protein that can be applied for various silica-based materials.

Keywords

biomimetic (chemical reaction)biomaterialenvironmentally benign
Type
Articles
Information
MRS Online Proceedings Library (OPL) , Volume 1569: Symposium LL – Advanced Materials for Biological and Biomedical Applications , 2013 , pp. 251 - 258
Copyright
Copyright © Materials Research Society 2013 

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References

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