Prolyl-specific peptidases for applications in food protein hydrolysis

Mika, Nicole; Zorn, Holger; Rühl, Martin

doi:10.1007/s00253-015-6838-0

Cited by 25 publications

(16 citation statements)

References 74 publications

(87 reference statements)

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“…S5 ). At 29 kDa, the enzyme is considerably smaller than any known prolyl endoproteases, and does not appear to have the substrate length restrictions observed with prolyl endopeptidases 43 . We have named this newly-discovered proteolytic enzyme neprosin (Npr1).…”

Section: Resultsmentioning

confidence: 82%

Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease

Rey

Yang

Lee

et al. 2016

Sci Rep

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Celiac disease is triggered by partially digested gluten proteins. Enzyme therapies that complete protein digestion in vivo could support a gluten-free diet, but the barrier to completeness is high. Current options require enzyme amounts on the same order as the protein meal itself. In this study, we evaluated proteolytic components of the carnivorous pitcher plant (Nepenthes spp.) for use in this context. Remarkably low doses enhance gliadin solubilization rates, and degrade gliadin slurries within the pH and temporal constraints of human gastric digestion. Potencies in excess of 1200:1 (substrate-to-enzyme) are achieved. Digestion generates small peptides through nepenthesin and neprosin, the latter a novel enzyme defining a previously-unknown class of prolyl endoprotease. The digests also exhibit reduced TG2 conversion rates in the immunogenic regions of gliadin, providing a twin mechanism for evading T-cell recognition. When sensitized and dosed with enzyme-treated gliadin, NOD/DQ8 mice did not show intestinal inflammation, when compared to mice challenged with only pepsin-treated gliadin. The low enzyme load needed for effective digestion suggests that gluten detoxification can be achieved in a meal setting, using metered dosing based on meal size. We demonstrate this by showing efficient antigen processing at total substrate-to-enzyme ratios exceeding 12,000:1.

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Section: Resultsmentioning

confidence: 82%

Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease

Rey

Yang

Lee

et al. 2016

Sci Rep

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“…This indicates that Clostridium sporogenes KAR95 produces mostly prolyl endopeptidase. The activity of prolyl endopeptidase is often restricted to substrates shorter than 30 amino acids and hydrolysis at a central position (Mika et al 2015 ). In conclusion, bacterial strains that originate from vegetable roots produce proteases with very different efficiency and target specificity.…”

Section: Discussionmentioning

confidence: 99%

Microbiome of root vegetables—a source of gluten-degrading bacteria

Kõiv

Adamberg

et al. 2020

Appl Microbiol Biotechnol

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Gluten is a cereal protein that is incompletely digested by human proteolytic enzymes that create immunogenic peptides that accumulate in the gastrointestinal tract (GIT). Although both environmental and human bacteria have been shown to expedite gluten hydrolysis, gluten intolerance is a growing concern. Here we hypothesize that together with food, we acquire environmental bacteria that could impact our GIT with gluten-degrading bacteria. Using in vitro gastrointestinal simulation conditions, we evaluated the capacity of endophytic bacteria that inhabit root vegetables, potato (Solanum tuberosum), carrot (Daucus sativus), beet (Beta vulgaris), and topinambur (Jerusalem artichoke) (Helianthus tuberosus), to resist these conditions and degrade gluten. By 16S rDNA sequencing, we discovered that bacteria from the families Enterobacteriaceae, Bacillaceae, and Clostridiaceae most effectively multiply in conditions similar to the human GIT (microoxic conditions, 37 °C) while utilizing vegetable material and gluten as nutrients. Additionally, we used stomach simulation (1 h, pH 3) and intestinal simulation (1 h, bile salts 0.4%) treatments. The bacteria that survived this treatment retained the ability to degrade gluten epitopes but at lower levels. Four bacterial strains belonging to species Bacillus pumilus, Clostridium subterminale, and Clostridium sporogenes isolated from vegetable roots produced proteases with postproline cleaving activity that successfully neutralized the toxic immunogenic epitopes. Key points • Bacteria from root vegetables can degrade gluten. • Some of these bacteria can resist conditions mimicking gastrointestinal tract.

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“…1 % der Weltbevölkerung leidet, hergestellt werden. Da Getreide‐ und Vorratsschädlinge wie der Getreideplattkäfer ( Oryzaephilus surinamensis ), der Getreidekapuziner ( Rhizopertha dominica ), der Kornkäfer ( Sitophilus granarius ) oder der Mehlkäfer ( Tenebrio molitor ) sich von Getreide, und damit Gluten, ernähren, müssen sie über Peptidasen zur Hydrolyse dieser für manche Menschen schwer verdaulichen Proteine verfügen .…”

Section: Gelbe Biotechnologie (Insektenbiotechnologie)unclassified

“…Im nächsten Schritt werden die Peptidasen entweder direkt mit einem sterilen Skalpell aus den Zymogrammen ausgeschnitten oder, um eine bessere Abtrennung von anderen Proteinen zu erreichen, chromatographisch (mittels fast protein liquid chromatography ; FPLC) gereinigt . Mit Hilfe massenspektrometrischer Verfahren werden Peptidsequenzen der Enzyme ermittelt und diese mit dem Transkriptom des betreffenden Käfers (Infokasten 2) abgeglichen (Abbildung ).…”

Section: Gelbe Biotechnologie (Insektenbiotechnologie)unclassified

“…Als Substrat zur selektiven Detektion von Endopeptidaseaktivität dient die Modell verbindung Benzyloxycarbonylglycyllprolinpnitroanilid (Abbildung 2). Durch die Peptidasekatalysierte Hydrolyse wird pNitroanilin freigesetzt, dessen Absorption bei 410 nm photometrisch verfolgt wird.Im nächsten Schritt werden die Peptidasen entweder direkt mit einem sterilen Skalpell aus den Zymogrammen ausgeschnitten oder, um eine bessere Abtrennung von ande ren Proteinen zu erreichen, chromatographisch (mittels fast protein liquid chromatography; FPLC) gereinigt[4,5].…”

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Insekten und Schimmelpilze als Bioressource

Rühl¹,

Pietzner²,

Vilcinskas³

et al. 2019

Chemie in nserer Zeit

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Zusammenfassung Mikroorganismen werden seit Jahrtausenden zur Herstellung von Lebensmitteln genutzt. Mittlerweile sind diese biotechnologischen Verfahren nicht mehr auf die Verwendung einiger Organismen beschränkt. Die Möglichkeit, Enzyme unabhängig von ihrem Wirtsorganismus zu produzieren, erweiterte den Einsatzbereich dieser technischen Hilfsmittel ungemein. Seither werden vermehrt neue Bioressourcen erschlossen, um neuartige Enzyme und Peptide für die Lebensmittelproduktion verfügbar zu machen: Peptidasen aus Getreideschädlingen vermögen Gluten zu hydrolysieren und antimikrobiell wirksame Peptide aus Insekten könnten zur Haltbarmachung von Lebensmitteln eingesetzt werden. Aber wie findet man solche Katalysatoren? Beispiele aus eigenen Arbeiten zeigen es.

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Prolyl-specific peptidases for applications in food protein hydrolysis

Cited by 25 publications

References 74 publications

Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease

Addressing proteolytic efficiency in enzymatic degradation therapy for celiac disease

Microbiome of root vegetables—a source of gluten-degrading bacteria

Insekten und Schimmelpilze als Bioressource

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