1887

Abstract

Prolyl endopeptidases are key enzymes in the digestion of proline-rich proteins. Fungal extracts rich in prolyl endopeptidases produced by a species such as used in food fermentation would be of particular interest for the development of an oral enzyme therapy product in patients affected by intolerance to gluten. Two major secreted prolyl endopeptidases of the MEROPS S28 peptidase family, AoS28A and AoS28B, were identified when this fungus was grown at acidic pH in a medium containing soy meal protein or wheat gliadin as the sole source of nitrogen. AoS28B was produced by 12 reference strains used in food fermentation. AoS28A was secreted by six of these 12 strains. This protease is the orthologue of the previously characterized (AfuS28) and (AN-PEP) prolyl endopeptidases which are encoded by genes with a similar intron–exon structure. Large amounts of secreted AoS28A and AoS28B were obtained by gene overexpression in . AoS28A and AoS28B are endoproteases able to cleave N-terminally blocked proline substrates. Both enzymes very efficiently digested the proline-rich 33-mer of gliadin, the most representative immunotoxic peptide deriving from gliadin, with some differences in terms of specificity and optimal pH. Digestion of the gliadin peptide in short peptides with both enzymes was found to occur from its N terminus.

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2015-12-01
2021-09-23
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References

  1. Beauvais A., Monod M., Wyniger J., Debeaupuis J. P., Grouzmann E., Brakch N., Svab J., Hovanessian A. G., Latgé J. P. (1997). Dipeptidyl-peptidase IV secreted by Aspergillus fumigatus, a fungus pathogenic to humansInfect Immun 6530423047[PubMed]. [Google Scholar]
  2. Bethune M. T., Strop P., Tang Y., Sollid L. M., Khosla C. (2006). Heterologous expression, purification, refolding, and structural-functional characterization of EP-B2, a self-activating barley cysteine endoproteaseChem Biol 13637647 [View Article][PubMed]. [Google Scholar]
  3. Blinkovsky A. M., Byun T., Brown K. M., Golightly E. J., Klotz A. V. (2000). A non-specific aminopeptidase from Aspergillus Biochim Biophys Acta 1480171181 [View Article][PubMed]. [Google Scholar]
  4. Borg-von Zepelin M., Beggah S., Boggian K., Sanglard D., Monod M. (1998). The expression of the secreted aspartyl proteinases Sap4 to Sap6 from Candida albicans in murine macrophagesMol Microbiol 28543554 [View Article][PubMed]. [Google Scholar]
  5. Byun T., Kofod L., Blinkovsky A. (2001). Synergistic action of an X-prolyl dipeptidyl aminopeptidase and a non-specific aminopeptidase in protein hydrolysisJ Agric Food Chem 4920612063 [View Article][PubMed]. [Google Scholar]
  6. Cawley N. X., Olsen V., Zhang C. F., Chen H. C., Tan M., Loh Y. P. (1998). Activation and processing of non-anchored yapsin 1 (Yap3p)J Biol Chem 273584591 [View Article][PubMed]. [Google Scholar]
  7. Chakraborty B. N., Patterson N. A., Kapoor M. (1991). An electroporation-based system for high-efficiency transformation of germinated conidia of filamentous fungiCan J Microbiol 37858863 [View Article][PubMed]. [Google Scholar]
  8. Chien H.-C. R., Lin L.-L., Chao S.-H., Chen C.-C., Wang W.-C., Shaw C.-Y., Tsai Y.-C., Hu H.-Y., Hsu W.-H. (2002). Purification, characterization, and genetic analysis of a leucine aminopeptidase from Aspergillus sojae Biochim Biophys Acta 1576119126 [View Article][PubMed]. [Google Scholar]
  9. Cove D. J. (1966). The induction and repression of nitrate reductase in the fungus Aspergillus nidulans Biochim Biophys Acta 1135156 [View Article][PubMed]. [Google Scholar]
  10. Doumas A., van den Broek P., Affolter M., Monod M. (1998). Characterization of the prolyl dipeptidyl peptidase gene (dppIV) from the koji mold Aspergillus oryzae Appl Environ Microbiol 6448094815[PubMed]. [Google Scholar]
  11. Edens L., Dekker P., van der Hoeven R., Deen F., de Roos A., Floris R. (2005). Extracellular prolyl endoprotease from Aspergillus niger and its use in the debittering of protein hydrolysatesJ Agric Food Chem 5379507957 [View Article][PubMed]. [Google Scholar]
  12. Kang C., Yu X.-W., Xu Y. (2015). Cloning and expression of a novel prolyl endopeptidase from Aspergillus oryzae and its application in beer stabilizationJ Ind Microbiol Biotechnol 42263272 [View Article][PubMed]. [Google Scholar]
  13. Kim C.-Y., Quarsten H., Bergseng E., Khosla C., Sollid L. M. (2004). Structural basis for HLA-DQ2-mediated presentation of gluten epitopes in celiac diseaseProc Natl Acad Sci U S A 10141754179 [View Article][PubMed]. [Google Scholar]
  14. Kitamoto N., Matsui J., Kawai Y., Kato A., Yoshino S., Ohmiya K., Tsukagoshi N. (1998). Utilization of the TEF1-alpha gene (TEF1) promoter for expression of polygalacturonase genes, pgaA and pgaB, in Aspergillus oryzae Appl Microbiol Biotechnol 508592 [View Article][PubMed]. [Google Scholar]
  15. Machida M., Asai K., Sano M., Tanaka T., Kumagai T., Terai G., Kusumoto K., Arima T., Akita O., other authors. (2005). Genome sequencing and analysis of Aspergillus oryzae Nature 43811571161 [View Article][PubMed]. [Google Scholar]
  16. MacKay V. L., Welch S. K., Insley M. Y., Manney T. R., Holly J., Saari G. C., Parker M. L. (1988). The Saccharomyces cerevisiae BAR1 gene encodes an exported protein with homology to pepsinProc Natl Acad Sci U S A 855559 [View Article][PubMed]. [Google Scholar]
  17. Monod M., Jousson O., Reichard U. (2009). Aspergillus fumigatus secreted proteases. In Aspergillus fumigatus and Aspergillosis, pp. 87106. Edited by Latgé J.-P., Steinbach W. J. Washington, DCAmerican Society for Microbiology.[CrossRef] [Google Scholar]
  18. Nikkuni S., Nakajima H., Hoshina S. I., Ohno M., Suzuki C., Kashiwagi Y., Mori K. (1998). Evolutionary relationships among Aspergillus oryzae and related species based on the sequences of 18S rRNA genes and internal transcribed spacersJ Gen Appl Microbiol 44225230 [View Article][PubMed]. [Google Scholar]
  19. Reichard U., Léchenne B., Asif A. R., Streit F., Grouzmann E., Jousson O., Monod M. (2006). Sedolisins, a new class of secreted proteases from Aspergillus fumigatus with endoprotease or tripeptidyl-peptidase activity at acidic pHsAppl Environ Microbiol 7217391748 [View Article][PubMed]. [Google Scholar]
  20. Sambrook J., Fritsch E. F., Maniatis T. (1989). Molecular Cloning: A Laboratory Manual2nd ednCold Spring Harbor, NYCold Spring Harbor Laboratory. [Google Scholar]
  21. Sarfati J., Monod M., Recco P., Sulahian A., Pinel C., Candolfi E., Fontaine T., Debeaupuis J. P., Tabouret M., Latgé J. P. (2006). Recombinant antigens as diagnostic markers for aspergillosisDiagn Microbiol Infect Dis 55279291 [View Article][PubMed]. [Google Scholar]
  22. Shan L., Molberg Ø., Parrot I., Hausch F., Filiz F., Gray G. M., Sollid L. M., Khosla C. (2002). Structural basis for gluten intolerance in celiac sprueScience 29722752279 [View Article][PubMed]. [Google Scholar]
  23. Siegel M., Bethune M. T., Gass J., Ehren J., Xia J., Johannsen A., Stuge T. B., Gray G. M., Lee P. P., Khosla C. (2006). Rational design of combination enzyme therapy for celiac sprueChem Biol 13649658 [View Article][PubMed]. [Google Scholar]
  24. Sollid L. M., Qiao S.-W. W., Anderson R. P., Gianfrani C., Koning F. (2012). Nomenclature and listing of celiac disease relevant gluten T-cell epitopes restricted by HLA-DQ moleculesImmunogenetics 64455460 [View Article][PubMed]. [Google Scholar]
  25. Sriranganadane D., Waridel P., Salamin K., Reichard U., Grouzmann E., Neuhaus J. M., Quadroni M., Monod M. (2010). Aspergillus protein degradation pathways with different secreted protease sets at neutral and acidic pHJ Proteome Res 935113519 [View Article][PubMed]. [Google Scholar]
  26. Staib P., Lermann U., Blass-Warmuth J., Degel B., Würzner R., Monod M., Schirmeister T., Morschhäuser J. (2008). Tetracycline-inducible expression of individual secreted aspartic proteases in Candida albicans allows isoenzyme-specific inhibitor screeningAntimicrob Agents Chemother 52146156 [View Article][PubMed]. [Google Scholar]
  27. Stepniak D., Spaenij-Dekking L., Mitea C., Moester M., de Ru A., Baak-Pablo R., van Veelen P., Edens L., Koning F. (2006). Highly efficient gluten degradation with a newly identified prolyl endoprotease: implications for celiac diseaseAm J Physiol Gastrointest Liver Physiol 291G621G629 [View Article][PubMed]. [Google Scholar]
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