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Aims:  The purpose of this paper was to screen candidate bacterial strains for the production of proteases suitable for application to the degradation of pathogenic forms of prion protein (PrPSc). This paper describes the biochemical characteristics and proteolytic activity of the isolated protease. Methods and Results:  After screening more than 200 bacterial proteases for keratinolytic activity, we identified a Bacillus stain that produced a protease exhibiting high‐degradation activity against a scrapie PrPSc. Sequence analysis indicated that this serine‐protease belonged to the Subtilisin family and had optimum pH and temperature ranges of 9–10 and 60–70°C. Western blotting analysis revealed that the protease was also capable of decomposing bovine spongiform encephalopathy‐infected brain homogenate. In addition, the protease was demonstrated to degrade dried PrPSc that had become firmly attached to a plastic surface considerably more effectively than proteinase K or PWD‐1, a previously reported keratinase. Conclusions:  These results indicate that the isolated protease exhibited higher activity for PrPSc degradation compared with other proteases examined. Significance and Impact of the Study:  This protease could be used under moderate conditions for the decontamination of precision instruments that are susceptible to PrPSc contamination.

Background: Congenital hearing loss is one of the most common sensory disorders, with 50-70% of cases attributable to genetic causes. Although recent advances in the identification of deafness genes have resulted in more accurate molecular diagnosis, leading to the better determination of suitable clinical interventions, difficulties remain with regard to clinical applications due to the extreme genetic heterogeneity of deafness. Aim: Toward more effective genetic testing, we adopted Massively Parallel DNA Sequencing (MPS) of target genes using an Ion PGMÔ system and an Ion AmpliSeqÔ panel to diagnose common mutations responsible for deafness and discover rare causative gene mutations. Before its clinical application, we investigated the accuracy of MPS-based genetic testing. Results: We compared the results of Invader assay-based genetic screening, the accuracy of which has already been verified in previous studies, with those of MPS-based genetic testing for a large population of Japanese deafness patients and revealed that over 99.98% of the results were the same for each genetic testing system. Conclusion: The Ion Personal Genome Machine system had sufficient uniformity and accuracy for application to the clinical diagnosis of common causative mutations and efficiently identified rare causative mutations and/or mutation candidates.

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