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

BMC Biotechnology

Azumi Hirata³  Yuki Hori³  Yuichi Koga²  Jun Okada³  Akikazu Sakudo¹  Kazuyoshi Ikuta⁴  Shigenori Kanaya²  Kazufumi Takano³ 

[1] Faculty of Medicine, University of the Ryukyus, Nishihara, Japan

[2] Department of Material and Life Science, Graduate School of Engineering, Osaka University, Suita, Japan

[3] Laboratory of Biological Chemistry, Department of Biomolecular Chemistry, Kyoto Prefectural University, Kyoto, Japan

[4] Department of Virology, Center for Infectious Disease Control, Research Institute for Microbial Diseases, Osaka University, Suita, Japan

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

Others  :  1123231
DOI  :  10.1186/1472-6750-13-19

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

【 摘 要 】

Background

Tk-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 (PrP^Sc), a protease-resistant isoform of normal prion protein (PrP^C).

Results

Tk-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 PrP^Sc to a level undetectable by western-blot analysis.

Conclusions

Our 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】

   
2013 Hirata et al; licensee BioMed Central Ltd.

【 图 表 】

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