Advertisement

European Food Research and Technology

, Volume 242, Issue 4, pp 585–597 | Cite as

Production and application of barley malt extract with high peptidase activity for the degradation of gluten in wort

  • Verena Knorr
  • Roland Kerpes
  • Herbert Wieser
  • Martin Zarnkow
  • Thomas Becker
  • Peter KoehlerEmail author
Original Paper

Abstract

Special malt with high gluten-specific peptidase activity was prepared by germinating barley grain for 8 days at 18 °C and 48 % humidity. Endogenous malt peptidases were characterized and used to degrade gluten in barley wort. Peptidases of aqueous malt extracts degraded celiac-active peptides by cleaving peptide bonds involving proline residues. The peptidase activity was not affected by cultivar, area of cultivation, and high temperatures up to 80 °C during kiln drying. Temperatures up to 50 °C were tolerated without loss of full activity in aqueous solution. The presence of ethanol decreased the activity significantly. Cross-flow filtration, freeze-drying, and evaporation under reduced pressure at 50 °C were used to produce a concentrated enzyme-active malt extract. The latter method yielded the best results, and the obtained concentrate was used to study gluten degradation in barley wort as affected by concentration of the extract, incubation time, and temperature. The gluten content was determined by a competitive R5-ELISA. The addition of 10 % concentrated extract (grist-to-water ratio 1:2.5; 40.4 °Brix) to wort and incubation for 24 h at 50 °C resulted in a gluten-free wort.

Keywords

Celiac disease Gluten Peptidase Barley Malt extract Beer 

Notes

Acknowledgments

This research project was supported by the German Ministry of Economics and Technology (via AiF) and the FEI (Forschungskreis der Ernährungsindustrie e.V., Bonn), project number AiF 16791N. The authors would furthermore like to thank Florian Gschwendtner for excellent assistance and Dr. Jean Titze from Doehler Group for providing the facilities and knowledge for the upscale.

Compliance with ethical statements

Conflict of interest

None.

Compliance with ethics requirements

This article does not contain any studies with human or animal subjects.

References

  1. 1.
    Maiuri L, Troncone R, Mayer M, Coletta S, Picarelli A, de Vincenzi M, Pavone V, Auricchio S (1996) In vitro activities of A-gliadin related synthetic peptides. Damaging effect on the atrophic coeliac mucosa and activation of mucosal immune response in treated coeliac mucosa. Scand J Gastroenterol 31:247–253CrossRefGoogle Scholar
  2. 2.
    Ellis HJ, Pollock EL, Engel W, Fraser JS, Rosen-Bronson S, Wieser H, Ciclitira PJ (2003) Investigation of the putative immunodominant T cell epitopes in coeliac disease. Gut 52:212–217CrossRefGoogle Scholar
  3. 3.
    Wieser H, Koehler P (2008) The biochemical basis of celiac disease. Cereal Chem 85:1–13CrossRefGoogle Scholar
  4. 4.
    Codex standard for foods for special dietary use for persons intolerant to gluten. Codex Alimentarius, International food standards. CODEX STAN 118-1979, adopted in 1979; amended 1983; revised 2008. Rome, ItalyGoogle Scholar
  5. 5.
    Wieser H, Koehler P (2012) Detoxification of gluten by means of enzymatic treatment. J AOAC Int 95:356–363CrossRefGoogle Scholar
  6. 6.
    van Landschoot A (2011) Gluten-free barley malt beers. Cerevisia 36:93–97CrossRefGoogle Scholar
  7. 7.
    Dostálek P, Hochel I, Méndez E, Hernando A, Gabrovská D (2006) Immunochemical determination of gluten in malts and beers. Food Addit Contam 23:1074–1078CrossRefGoogle Scholar
  8. 8.
    Hartman G, Koehler P, Wieser H (2006) Rapid degradation of gliadin peptides active for coeliac disease patients by proteases from germinating cereals. J Cereal Sci 44:368–371CrossRefGoogle Scholar
  9. 9.
    Gessendorfer B, Koehler P, Wieser H (2009) Preparation and characterization of enzymatically hydrolyzed prolamins from wheat, rye, and barley as references for the immunochemical quantitation of partially hydrolyzed gluten. Anal Bioanal Chem 395:1721–1728CrossRefGoogle Scholar
  10. 10.
    Kerpes R, Knorr V, Procopio S, Koehler P, Becker T (2015) Gluten-specific peptidase activity of barley as affected by malting and its impact on gluten degradation. J Cereal Sci (submitted) Google Scholar
  11. 11.
    MEBAK (2006) Mitteleuropäische Brautechnische Analysenkommission. Collection of brewing analysis methods of the Mitteleuropäische Brautechnische Analysenkommission (MEBAK). Barley, adjuncts, malt, hops and hop products. MEBAK, Freising, GermanyGoogle Scholar
  12. 12.
    Gessendorfer B, Hartmann G, Wieser H, Koehler P (2011) Determination of celiac disease-specific peptidase activity of germinated cereals. Eur Food Res Technol 232:205–209CrossRefGoogle Scholar
  13. 13.
    Schwalb T, Wieser H, Koehler P (2012) Studies on the gluten-specific peptidase activity of germinated grains from different cereal species and cultivars. Eur Food Res Technol 235:1161–1170CrossRefGoogle Scholar
  14. 14.
    Baenziger PS, Clements RL, McIntosh MS, Yamazaki WT, Starling TM, Sammons DJ, Johnson JW (1985) Effect of cultivar, environment, and their interaction and stability analyses on milling and baking quality of soft red winter wheat. Crop Sci 25:5–8CrossRefGoogle Scholar
  15. 15.
    de Ritis G, Auricchio S, Jones HW, Lew EJ-L, Bernardin JE, Kasarda DD (1988) In-vitro (organ culture) studies of the activity of specific A-gliadin peptides in celiac disease. Gastroenterology 94:41–49Google Scholar
  16. 16.
    Wieser H, Belitz H-D, Ashkenazi A (1984) Amino-acid sequence of the coeliac active gliadin peptide B 3142. Eur Food Res Technol 179:371–376Google Scholar
  17. 17.
    Beal AD, Mottram DS (1993) An evaluation of the aroma characteristics of malted barley by free-choice profiling. J Sci Food Agric 61:17–22CrossRefGoogle Scholar
  18. 18.
    Wieser H, Belitz H-D, Idar D, Ashkenazi A (1986) Coeliac activity of the gliadin peptides CT-1 and CT-2. Eur Food Res Technol 182:115–117Google Scholar
  19. 19.
    Shan L, Molberg Ø, Parrot I, Hausch F, Filiz F, Gray GM, Sollid LM, Khosla C (2002) Structural basis for gluten intolerance in celiac sprue. Science 297:2275–2279CrossRefGoogle Scholar
  20. 20.
    Qiao SW, Bergseng E, Molberg O, Jung G, Fleckenstein B, Sollid LM (2005) Refining the rules of gliadin T cell epitope binding to the disease-associated DQ2 molecule in celiac disease: importance of proline spacing and glutamine deamidation. J Immunol 175:254–261CrossRefGoogle Scholar
  21. 21.
    Fraser JS, Enoel W, Ellis HJ, Moodie SJ, Pollock EL, Wieser H, Ciclitira PJ (2003) Coeliac disease in vivo activity of the putative immunodominant epitope. Gut 52:1698–1702CrossRefGoogle Scholar
  22. 22.
    Arentz-Hansen EH, McAdam SN, Molberg O, Kristiansen C, Sollid LM (2000) Production of a panel of recombinant gliadins for the characterisation of T cell reactivity in coeliac disease. Gut 46:46–51CrossRefGoogle Scholar
  23. 23.
    Arentz-Hansen H, Körner R, Molberg O, Quarsten H, Vader W, Kooy YM, Lundin KE, Koning F, Roepstorff P, Sollid LM, McAdam SN (2000) The intestinal T cell response to gliadin in adult celiac disease is focused on a single deamidated glutamine targeted by tissue transglutaminase. J Exp Med 191:603–612CrossRefGoogle Scholar
  24. 24.
    Arentz-Hansen H, McAdam SN, Molberg Ø, Fleckenstein B, Lundin KE, Jørgensen TJ, Jung G, Roepstorff P, Sollid LM (2002) Celiac lesion T cells recognize epitopes that cluster in regions of gliadins rich in proline residues. Gastroenterology 123:803–809CrossRefGoogle Scholar
  25. 25.
    Vader W, Kooy Y, Van Veelen P, De Ru A, Harris D, Benckhuijsen W, Peña S, Mearin L, Drijfhout JW, Koning F (2002) The gluten response in children with celiac disease is directed toward multiple gliadin and glutenin peptides. Gastroenterology 122:1729–1737CrossRefGoogle Scholar
  26. 26.
    Vader LW, Stepniak DT, Bunnik EM, Kooy YMC, de Haan W, Drijfhout JW, van Veelen PA, Koning F (2003) Characterization of cereal activity for celiac disease patients based on protein homology in grains. Gastroenterology 125:1105–1113CrossRefGoogle Scholar
  27. 27.
    Tye-Din JA, Stewart JA, Dromey JA, Beissbarth T, van Heel DA, Tatham A, Henderson K, Mannering SI, Gianfrani C, Jewell DP, Hill AV, McCluskey J, Rossjohn J, Anderson RP (2010) Comprehensive, quantitative mapping of T cell epitopes in gluten in celiac disease. Sci Transl Med 2:41–51CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Verena Knorr
    • 1
  • Roland Kerpes
    • 2
  • Herbert Wieser
    • 1
  • Martin Zarnkow
    • 3
  • Thomas Becker
    • 2
  • Peter Koehler
    • 1
    Email author
  1. 1.Deutsche Forschungsanstalt für LebensmittelchemieLeibniz InstitutFreisingGermany
  2. 2.Lehrstuhl für Brau- und GetränketechnologieTechnische Universität MünchenFreisingGermany
  3. 3.Forschungszentrum Weihenstephan für Brau- und LebensmittelqualitätTechnische Universität MünchenFreisingGermany

Personalised recommendations