Enzymatic activity of a subtilisin homolog, Tk-SP, from Thermococcus kodakarensisin detergents and its ability to degrade the abnormal prion protein

BMC Biotechnology

Research Article

Yuichi Koga¹  Shigenori Kanaya¹  Kazuyoshi Ikuta²  Akikazu Sakudo³  Kazufumi Takano⁴  Jun Okada⁴  Azumi Hirata⁴  Yuki Hori⁴ 

[1] Department of Material and Life Science, Graduate School of Engineering, Osaka University, Suita, Japan;Department of Virology, Center for Infectious Disease Control, Research Institute for Microbial Diseases, Osaka University, Suita, Japan;Faculty of Medicine, University of the Ryukyus, Nishihara, Japan;Laboratory of Biological Chemistry, Department of Biomolecular Chemistry, Kyoto Prefectural University, Kyoto, Japan;

关键词: Serine protease;    Hyperthermophilic archaeon;    Subtilisin;    Detergent compatibility;    Prion;    Transmissible spongiform encephalopathies (TSE);    Degradation;    Decontamination;   

DOI  :  10.1186/1472-6750-13-19

 received in 2012-11-19, accepted in 2013-02-26,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundTk-SP is a member of subtilisin-like serine proteases from a hyperthermophilic archaeon Thermococcus kodakarensis. It has been known that the hyper-stable protease, Tk-SP, could exhibit enzymatic activity even at high temperature and in the presence of chemical denaturants. In this work, the enzymatic activity of Tk-SP was measured in the presence of detergents and EDTA. In addition, we focused to demonstrate that Tk-SP could degrade the abnormal prion protein (PrPSc), a protease-resistant isoform of normal prion protein (PrPC).ResultsTk-SP was observed to maintain its proteolytic activity with nonionic surfactants and EDTA at 80°C. We optimized the condition in which Tk-SP functions efficiently, and demonstrated that the enzyme is highly stable in the presence of 0.05% (w/v) nonionic surfactants and 0.01% (w/v) EDTA, retaining up to 80% of its activity. Additionally, we also found that Tk-SP can degrade PrPSc to a level undetectable by western-blot analysis.ConclusionsOur results indicate that Tk-SP has a great potential for technological applications, such as thermo-stable detergent additives. In addition, it is also suggested that Tk-SP-containing detergents can be developed to decrease the secondary infection risks of transmissible spongiform encephalopathies (TSE).

【 授权许可】

Unknown   
© Hirata et al; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.