Identification of an alkaliphilic actinomycetes that produces a PrPSc-degrading enzyme

Mitsuiki, Shinji; Takasugi, Masayuki; Moriyama, Yasushi; Futagami, Taiki; Goto, Minoru; Kanouchi, Hiroaki; Oka, Tatsuzo

doi:10.1007/s13213-010-0049-9

Cited by 9 publications

(5 citation statements)

References 27 publications

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“…The vector was described in a paper by Piening et al [48]. RK13 VRQ.G9 cells have similar level of sensitivity as Rov9 cells [49] to SSBP/1 infection (Neale, unpublished data) and were used in the standard scrapie cell assay (SSCA). The cells were maintained in Eagle’s minimal essential medium (EMEM, Gibco) supplemented with 10% fetal calf serum and 2% HEPES.…”

Section: Methodsmentioning

confidence: 99%

Enzymatic Formulation Capable of Degrading Scrapie Prion under Mild Digestion Conditions

et al. 2013

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The prion agent is notoriously resistant to common proteases and conventional sterilisation procedures. The current methods known to destroy prion infectivity such as incineration, alkaline and thermal hydrolysis are harsh, destructive, environmentally polluting and potentially hazardous, thus limit their applications for decontamination of delicate medical and laboratory devices, remediation of prion contaminated environment and for processing animal by-products including specified risk materials and carcases. Therefore, an environmentally friendly, non-destructive enzymatic degradation approach is highly desirable. A feather-degrading Bacillus licheniformis N22 keratinase has been isolated which degraded scrapie prion to undetectable level of PrPSc signals as determined by Western Blot analysis. Prion infectivity was verified by ex vivo cell-based assay. An enzymatic formulation combining N22 keratinase and biosurfactant derived from Pseudomonas aeruginosa degraded PrPSc at 65°C in 10 min to undetectable level -. A time-course degradation analysis carried out at 50°C over 2 h revealed the progressive attenuation of PrPSc intensity. Test of residual infectivity by standard cell culture assay confirmed that the enzymatic formulation reduced PrPSc infectivity to undetectable levels as compared to cells challenged with untreated standard scrapie sheep prion (SSBP/1) (p-value = 0.008 at 95% confidence interval). This novel enzymatic formulation has significant potential application for prion decontamination in various environmentally friendly systems under mild treatment conditions.

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Section: Methodsmentioning

confidence: 99%

Enzymatic Formulation Capable of Degrading Scrapie Prion under Mild Digestion Conditions

et al. 2013

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“…When it was used in the paper industry, the quality of paper could be improved (Jagannathan et al 2021;Angural et al 2020). In addition, alkaline enzymes produced by alkaliphilic Actinomycetes could also be used in sewage treatment, food, textiles, and other areas (Mitsuiki et al 2010). Therefore, the discovery of alkaliphilic Actinomycetes in extreme environments plays an important role in nding new bacterial resources and synthesizing new lead compounds.…”

Section: Introductionmentioning

confidence: 99%

Characterization and identification of filamentous bacterium XJ-16 and analysis of its bioactive components

Jiang

Bai

Luo

et al. 2023

Preprint

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Actinomycetes, which can produce a variety of bioactive compounds in the metabolic process, is one of the important sources of novel drugs, enzymes, anti-tumor drugs and enzyme inhibitors. It has been the focus of researchers to find and develop Actinomycetes with special characters. Strain XJ-16 is a blue alkali-resistant filamentous bacterium with high antimicrobial activity isolated from saline-alkali land of Xinjiang. Based on the classification, the enzyme production, metabolite antibacterial activity, and antibacterial substance isolation of XJ-16 were explored. which showed that XJ-16 belongs to the blue group of Streptomyces sp, and it can secrete cellulase, lipase, urease, protease, catalase and oxidase during metabolism. In addition, the bacteriostatic substance secreted by the strain XJ-16 showed inhibitory effects against both Gram-positive and Gram-negative bacteria, as well as the yeast Candida albicans. Then it was found that the bacteriostasis produced by XJ-16 has strong tolerance to acid, weak tolerance to alkali, and easy to be inactivated. After tested by HPLC, the retention time of antimicrobial substance was 13.261 min. This study provides a theoretical basis for the search for new antibacterial compounds and the further development of blue alkaline Actinomycetes XJ-16.

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“…Microbial keratinases are proteases that effectively degrade sturdy proteins such as keratin [20]. Keratinases are produced by several microbial species including fungi [21,22], actinomycetes [23] especially genus Streptomyces [24,25], Bacillus [26,27], and archaea [28]. A keratinase purified from Bacillus licheniformis PWD‐1 degraded prions efficiently [29].…”

Section: Introductionmentioning

confidence: 99%

In vitro degradation of β‐amyloid fibrils by microbial keratinase

Ningthoujam

Mukherjee

Devi

et al. 2019

A&D Transl Res & Clin Interv

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Introduction Amyloid fibrils are misfolded, protease-resistant forms of normal proteins. They are infectious such as prions or noninfectious such as β-amyloid (Aβ) fibrils causing Alzheimer's disease (AD). Prions and amyloids are structurally similar, possessing cross β-pleated sheet-like structures. As microbial keratinase could degrade prions, we tested keratinase activity on Aβ fibrils. Methods Lysozyme treated with urea generates Aβ fibrils demonstrated by immunoblotting with anti-Aβ antibody, high-performance liquid chromatography, and Congo red absorption spectroscopy. Two keratinases, Ker1 and Ker2, were purified from an actinomycete Amycolatopsis sp. MBRL 40 and incubated with Aβ fibrils. Results Soluble Ker1 and Ker1 reconstituted on neutral/cationic liposomes degraded Aβ fibrils efficiently. Ker 2 was less potent. Discussion Drugs that target AD inhibit acetylcholinesterase or formation of Aβ fibrils and downstream effects. These drugs have side effects and do not benefit globally in cognition. Keratinases are novel molecules for drug development against AD.

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Identification of an alkaliphilic actinomycetes that produces a PrPSc-degrading enzyme

Cited by 9 publications

References 27 publications

Enzymatic Formulation Capable of Degrading Scrapie Prion under Mild Digestion Conditions

Enzymatic Formulation Capable of Degrading Scrapie Prion under Mild Digestion Conditions

Characterization and identification of filamentous bacterium XJ-16 and analysis of its bioactive components

In vitro degradation of β‐amyloid fibrils by microbial keratinase

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