Alkaline serine protease produced by Streptomyces sp. degrades PrPSc

Zhao, Hui; Doi, Hiroyasu; Kanouchi, Hiroaki; Matsuura, Yuichi; Mohri, Shiro; Nonomura, Yoshiaki; Oka, Tatsuzo

doi:10.1016/j.bbrc.2004.06.100

Cited by 44 publications

(33 citation statements)

References 22 publications

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“…McKinley et al [11] reported that proteinase K digestion for more than 2 h leads to hydrolysis. In recent publications, thermostable alkaline serine proteases of Thermococcus, Thermosipho, Thermoanaerobacter and Streptomyces were able to degrade PrP Sc under special conditions [7,12,21]. Denaturation leads to a conformation of PrP Sc susceptible to proteolysis.…”

Section: Discussionmentioning

confidence: 99%

“…A 20% homogenate of the hamster brain tissue was prepared in homogenisation buffer (0.32 M sucrose, 0.5% sodium desoxycholate, 0.5% Nonidet P 40, pH 7) • C until use. All experiments were carried out in agreement with the national guidelines in a class 3** laboratory of the institute.…”

Section: Experimental Materialsmentioning

confidence: 99%

“…PrP Sc is resistant towards other proteolytic enzymes like trypsin, V8 protease [11] and pepsin [8]. Recently, data were published indicating that extra cellular proteases of Streptomyces, Bacillus and anaerobic thermophilic prokaryotes are able to decrease the PrP Sc level in infected brain homogenates of mice and hamsters [7,8,11,12]. Thus, TSE agents are sensitive towards proteases, although such proteases rarely appear to occur.…”

Section: Introductionmentioning

confidence: 99%

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Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Scherbel¹,

Pichner²,

Groschup

et al. 2006

Vet. Res.

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-A food-borne origin of the transmission of bovine spongiform encephalopathy (BSE) to cattle is commonly assumed. However, the fate of infectious prion protein during polygastric digestion remains unclear. It is unknown at present, whether infectious prion proteins, considered to be very stable, are degraded or inactivated by microbial processes in the gastrointestinal tract of cattle. In this study, rumen and colon contents from healthy cattle, taken immediately after slaughter, were used to assess the ability of these microbial consortia to degrade PrP Sc . Therefore, the consortia were incubated with brain homogenates of scrapie (strain 263K) infected hamsters under physiological anaerobic conditions at 37• C. Within 20 h, PrP Sc was digested both with ruminal and colonic microbiota up to immunochemically undetectable levels. Especially polymyxin resistant (mainly gram-positive) bacteria expressed PrP Sc degrading activity. These data demonstrate the ability of bovine gastrointestinal microbiota to degrade PrP Sc during digestion.transmissible spongiform encephalopathy / prion / degradation / microbiota / gastrointestinal tract

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Section: Discussionmentioning

confidence: 99%

Section: Experimental Materialsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Scherbel¹,

Pichner²,

Groschup

et al. 2006

Vet. Res.

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“…The subtilisin-enzyme Properase is reported to be able to degrade PrP Sc at high temperatures under alkaline conditions (18), while a keratinase from Bacillus licheniformis requires >100˚C and the presence of a detergent to break down PrP Sc (19). An alkaline protease, E77, derived from Streptmyces species has shown degradation of PrP Sc at 60˚C and pH 11.0 (20). Hamo 100 and Properase were proved to eliminate prion infectivity (18,21).…”

Section: ------------------------------------------------------------mentioning

confidence: 99%

Degradation and destabilization of abnormal prion protein using alkaline detergents and proteases

Sakudo¹

2009

Int J Mol Med

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Abstract. There is a limited number of reports regarding detergents and proteases inactivating, degrading, or destabilizing abnormal prion protein (PrP Sc ). In the present study, the effect of alkaline detergents and proteases on the breakdown of PrP Sc in the absence of proteinase K (PK) (degradation) and the presence of PK (destabilization) was investigated. PrPSc from brain homogenate of terminallydiseased mice infected with the Chandler strain of scrapie was used as a substrate. A surfactant-free alkaline detergent (pH 11.9, 1% aqueous solution) with potassium hydroxide as the main ingredient and an alkaline detergent (pH 11.9, 1% aqueous solution) containing about 1% surfactant as well as two commercially available alkaline proteases had a destabilizing effect on PrP Sc . All these detergents and proteases showed degradative effects on PrP Sc under appropriate conditions. These results demonstrate the usefulness of alkaline detergents and proteases for the degradation or destabilization of PrP Sc .

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“…Protease digestion and elimination of ELISA or Western-blot immunoreactivity has been shown under a variety of conditions (Hui et al, 2004a(Hui et al, , 2004bLangeveld et al, 2003;Tsiroulnikov et al, 2004;Mitsuiki et al, 2006;Rapp et al, 2006;Scherbel et al, 2006;Muller-Hellwig et al, 2006). However, protease destruction of prions below in vitro assay detection limits does not necessarily mean that PrP res has been reduced to levels that are no longer infectious in more sensitive bioassays; therefore, methods investigating decontamination of infectious prions need to be tested in vivo until better in vitro methods are developed and validated (McLeod et al, 2004;Lawson et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Feasibility of infectious prion digestion using mild conditions and commercial subtilisin

Pilon

Nash²,

Arver³

et al. 2009

Journal of Virological Methods

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a b s t r a c tTwo serine protease enzymes, subtilisin 309 and subtilisin 309-v, were used to digest brain homogenates containing high levels of prion infectivity using mildly alkaline conditions to investigate prion decontamination methods. To establish that PrP res infectivity was eliminated, we utilized the Rocky Mountain Laboratory (RML) mouse-adapted scrapie model system for bioassay. Only one digestion condition (subtilisin 309 at 138 mAU/ml, 55• C and 14 h digestion time pH 7.9) was considered to be highly relevant statistically (P < 0.001) compared to control, with 52% of challenged mice surviving until the end of the study period. In contrast, treatment of PrP res by autoclaving at 134• C or treatment with hypochlorite at a concentration of 20,000 ppm completely protected mice from prionosis. Further, in vitro assays suggest that potential proteolytic based PrP res decontamination methods must use high enzyme concentration, pH values >9.0, and elevated temperatures to be a safely efficacious, thereby limiting applicability on delicate surgical instruments and use in the environment.Published by Elsevier B.V.

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Alkaline serine protease produced by Streptomyces sp. degrades PrPSc

Cited by 44 publications

References 22 publications

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Degradation and destabilization of abnormal prion protein using alkaline detergents and proteases

Feasibility of infectious prion digestion using mild conditions and commercial subtilisin

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Alkaline serine protease produced by Streptomyces sp. degrades PrPSc

Cited by 44 publications

References 22 publications

Degradation of scrapie associated prion protein (PrPSc) by the gastrointestinal microbiota of cattle

Degradation of scrapie associated prion protein (PrPSc) by the gastrointestinal microbiota of cattle

Degradation and destabilization of abnormal prion protein using alkaline detergents and proteases

Feasibility of infectious prion digestion using mild conditions and commercial subtilisin

Contact Info

Product

Resources

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Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle

Degradation of scrapie associated prion protein (PrP^Sc) by the gastrointestinal microbiota of cattle