Targeted modification of wheat grain protein to reduce the content of celiac causing epitopes

Functional and Integrative Genomics

Volumn 12, Issue 3, 2012, Pages 417-438

  Osorio, C ^a   Wen, N ^a   Gemini, R ^a   Zemetra, R ^b   Von Wettstein, D ^a   Rustgi, S ^a  

^a WASHINGTON STATE UNIVERSITY   (United States)

^b OREGON STATE UNIVERSITY   (United States)

Author keywords

Combined enzyme therapy; Cysteine endoprotease; Gluten; Prolyl endopeptidase; Wheat

Indexed keywords

ACYLTRANSFERASE INHIBITOR; CYSTEINE PROTEINASE; EPITOPE; GLUTEN; GRAIN PROTEIN; HLA DQ2 ANTIGEN; LARAZOTIDE; NEXVAX2; PROLAMIN; PROLYL ENDOPEPTIDASE; PROTEIN GLUTAMINE GAMMA GLUTAMYLTRANSFERASE INHIBITOR; RECOMBINANT ENDOPROTEINASE B2; RECOMBINANT ENZYME; UNCLASSIFIED DRUG; VACCINE; VEGETABLE PROTEIN;

ADAPTIVE IMMUNITY; AMINO ACID SEQUENCE; BARLEY; CELIAC DISEASE; CEREAL; CHRYSEOBACTERIUM MENINGOSEPTICUM; CLINICAL TRIAL (TOPIC); CONFORMATIONAL TRANSITION; CYTOKINE PRODUCTION; DRUG EFFICACY; DRUG HALF LIFE; DRUG MECHANISM; DRUG TOLERABILITY; ENZYME ACTIVATION; ENZYME ACTIVE SITE; ENZYME ACTIVITY; ENZYME SPECIFICITY; ENZYME STRUCTURE; ENZYME SYNTHESIS; ENZYME THERAPY; GENE INTERACTION; GENETIC ASSOCIATION; GENETIC PREDISPOSITION; GENOTYPE ENVIRONMENT INTERACTION; GLUTEN FREE DIET; HUMAN; IMMUNE RESPONSE; IMMUNOGENICITY; IMMUNOLOGICAL TOLERANCE; NONHUMAN; PATHOGENESIS; PLANT GENOME; PRIORITY JOURNAL; PROTEIN MODIFICATION; REVIEW; RYE; SINGLE NUCLEOTIDE POLYMORPHISM; VIRTUAL REALITY; WHEAT;

ANTIGENS, PLANT; CELIAC DISEASE; CYSTEINE ENDOPEPTIDASES; DRUG THERAPY, COMBINATION; ENDOPEPTIDASES; EPITOPES; FLAVOBACTERIACEAE; GLUTENS; HLA-DQ ANTIGENS; HORDEUM; HUMANS; SEED STORAGE PROTEINS; SEEDS; TRITICUM;

ELIZABETHKINGIA MENINGOSEPTICA; HORDEUM; SECALE CEREALE; TRITICUM AESTIVUM;

EID: 84870299936     PISSN: 1438793X     EISSN: 14387948     Source Type: Journal    
DOI: 10.1007/s10142-012-0287-y     Document Type: Review

References (171)

1. Adamovic S, Amundsen SS, Lie BA, Gudjonsdottir AH, Ascher H, Ek J, van Heel DA, Nilsson S, Sollid LM, Torinsson Naluai A (2008) Association study of IL2/IL21 and FcgRIIa: significant association with the IL2/IL21 region in Scandinavian coeliac disease families. Genes Immun 9:364-367
2. Arentz-Hansen H, Körner R, Molberg Ø, Quarsten H, Vader W, Kooy Y, Lundin K, Koning F, Roepstorff P, Sollid L, McAdam S (2000) The intestinal T cell response to α-gliadin in adult celiac disease is focused on a single deamidated glutamine targeted by tissue transglutaminase. J Exp Med 191:603-612
3. Arentz-Hansen H, McAdam SN, Molberg Ø, Fleckenstein B, Lundin KEA, Jørgensen TJD, Jung G, Roepstorff P, Sollid LM (2002) Celiac lesion T cells recognize epitopes that cluster in regions of gliadins rich in proline residues. Gastroenterology 123:803-809
4. Becker D, Folck A, Knies P, Lörz H, Wieser H (2006) Silencing the α- gliadins in hexaploid bread wheat. In: Lookhart GL, Ng PKW (eds) Gluten proteins. AACC, St Paul
5. Bethune MT, Strop P, Tang Y, Sollid LM, Khosla C (2006) Heterologous expression, purification, refolding, and structuralfunctional characterization of EP-B2, a self-activating barley cysteine endoprotease. Chem Biol 13:637-647
6. Bourgey M, Calcagno G, Tinto N, Gennarelli D, Margaritte-Jeannin P, Greco L, Limongelli MG, Esposito O, Marano C, Troncone R, Spampanato A, Clerget-Darpoux F, Sacchetti L (2007) HLA related genetic risk for coeliac disease. Gut 56:1054-1059
7. Brandt I, De Vriendt K, Devreese B, Van Beeumen J, Van Dongen W, Augustyns K, De Meester I, Scharpe S, Lambeir AM (2005) Search for substrates for prolyl oligopeptidase in porcine brain. Peptides 26:2536-2546
8. Branlard G, Dardevet M, Saccomano R, Lagoutte F, Gourdon J (2001) Genetic diversity of wheat storage proteins and bread wheat quality. Euphytica 119:59-67
9. Brew-Appiah RAT, Rustgi S, Claar M, Langen G, Kogel K-H, Weigel D, von Wettstein D (2010) Artificial microRNAs for silencing wheat proteins causing celiac disease. Paper presented at the Joint Annual Meeting American Society of Plant Biologists and Canadian Society of Plant Physiology 2010, Montreal, Canada
10. Cerf-Bensussan N, Matysiak-Budnik T, Cellier C, Heyman M (2007) Oral proteases: a new approach to managing coeliac disease. Gut 56:157-160
11. Cermak T, Doyle EL, Christian M, Wang L, Zhang Y, Schmidt C, Baller JA, Somia NV, Bogdanove AJ, Voytas DF (2011) Efficient design and assembly of custom TALEN and other TAL effectorbased constructs for DNA targeting. Nucl Acids Res 39:e82
12. Collado MC, Calabuig M, Sanz Y (2007) Differences between the fecal microbiota of coeliac infants and healthy controls. Curr Issues Intest Microbiol 8:9-14
13. Collado M, Donat E, Ribes-Koninckx C, Calabuig M, Sanz Y (2008) Imbalances in faecal and duodenal Bifidobacterium species composition in active and non-active coeliac disease. BMC Microbiol 8:232
14. Collado MC, Donat E, Ribes-Koninckx C, Calabuig M, Sanz Y (2009) Specific duodenal and faecal bacterial groups associated with paediatric coeliac disease. J Clin Pathol 62:264-269
15. Cornell HJ, Macrae FA, Melny J, Pizzey CJ, Cook F, Mason S, Bhathal PS, Stelmasiak T (2005) Enzyme therapy for management of coeliac disease. Scand J Gastroenterol 40:1304-1312
16. Darewicz M, Dziuba J, Minkiewicz P (2007) Computational characterisation and identification of peptides for in silico detection of potentially celiac-toxic proteins. Food Sci Tech Int 13:125-133
17. D'Argenio G, Petrosino S, Gianfrani C, Valenti M, Scaglione G, Grandone I, Nigam S, Sorrentini I, Mazzarella G, Di Marzo V (2007) Overactivity of the intestinal endocannabinoid system in celiac disease and in methotrexate-treated rats. J Mol Med (Berl) 85:523-530
18. Darkoh C, El-Bouhssini M, Baum M, Clack B (2010) Characterization of a prolyl endoprotease from Eurygaster integriceps puton (Sunn pest) infested wheat. Arch Insect Biochem Physiol 74:163-178
19. Davy A, Svendsen I, Sorensen SO, Sorensen MB, Rouster J, Meldal M, Simpson DJ, Cameron-Mills V (1998a) Substrate specificity of barley cysteine endoproteases EP-A and EP-B. Plant Physiol 117:255-261
20. Davy A, Svendsen I, Sørensen SO, Sørensen MB, Rouster J, Meldal M, Simpson DJ, Cameron-Mills V (1998b) Substrate Specificity of Barley Cysteine Endoproteases EP-A and EP-B1. Plant Physiol 117:255-261
21. Davy A, Sorensen MB, Svendsen I, Cameron-Mills V, Simpson DJ (2000) Prediction of protein cleavage sites by the barley cysteine endoproteases EP-A and EP-B based on the kinetics of synthetic peptide hydrolysis. Plant Physiol 122:137-145
22. De Paolo RW, Abadie V, Tang F, Fehlner-Peach H, Hall JA, Wang W, Marietta EV, Kasarda DD, Waldmann TA, Murray JA, Semrad C, Kupfer SS, Belkaid Y, Guandalini S, Jabri B (2011) Co-adjuvant effects of retinoic acid and IL-15 induce inflammatory immunity to dietary antigens. Nature 471:220-224
23. Dema B, Martinez A, Fernandez-Arquero M, Maluenda C, Polanco I, de la Concha EG, Urcelay E, Nunez C (2009) Association of IL18RAP and CCR3 with coeliac disease in the Spanish population. J Med Genet 46:617-619
24. Dewar D, Pereira SP, Ciclitira PJ (2004) The pathogenesis of coeliac disease. Int J Biochem Cell Biol 36:17-24
25. Dieterich W, Ehnis T, Bauer M, Donner P, Volta U, Riecken EO, Schuppan D (1997) Identification of tissue transglutaminase as the autoantigen of celiac disease. Nat Med 3:797-801
26. Djilali-Saiah I, Schmitz J, Harfouch-Hammoud E, Mougenot J-F, Bach J-F, Caillat-Zucman S (1998) CTLA-4 gene polymorphism is associated with predisposition to coeliac disease. Gut 43:187-189
27. Domenico L, Donald K (1985) One and two dimensional (two pH) polyacrilamide gel electrophoresis in a single gel: separation of wheat proteins. Cereal Chem 62:314-319
28. Drago S, El Asmar R, Di Pierro M, Grazia Clemente M, Tripathi A, Sapone A, Thakar M, Iacono G, Carroccio A, D'Agate C, Not T, Zampini L, Catassi C, Fasano A (2006) Gliadin, zonulin and gut permeability: effects on celiac and non-celiac intestinal mucosa and intestinal cell lines. Scand J Gastroenterol 41:408-419
29. Dubois PC, Trynka G, Franke L, Hunt KA, Romanos J, Curtotti A, Zhernakova A, Heap GA, Adany R, Aromaa A, Bardella MT, van den Berg LH, Bockett NA, de la Concha EG, Dema B, Fehrmann RS, Fernandez-Arquero M, Fiatal S, Grandone E, Green PM, Groen HJ, Gwilliam R, Houwen RH, Hunt SE, Kaukinen K, Kelleher D, Korponay-Szabo I, Kurppa K, MacMathuna P, Maki M, Mazzilli MC, McCann OT, Mearin ML, Mein CA, Mirza MM, Mistry V, Mora B, Morley KI, Mulder CJ, Murray JA, Nunez C, Oosterom E, Ophoff RA, Polanco I, Peltonen L, Platteel M, Rybak A, Salomaa V, Schweizer JJ, Sperandeo MP, Tack GJ, Turner G, Veldink JH, Verbeek WH, Weersma RK, Wolters VM, Urcelay E, Cukrowska B, Greco L, Neuhausen SL, McManus R, Barisani D, Deloukas P, Barrett JC, Saavalainen P, Wijmenga C, van Heel DA (2010) Multiple common variants for celiac disease influencing immune gene expression. Nat Genet 42:295-302
30. Farrel R, Kelly C (2002) Celiac sprue. N Engl J Med 346:180-188
31. Fleckenstein B, Molberg O, Qiao SW, Schmid DG, von der Mulbe F, Elgstoen K, Jung G, Sollid LM (2002) Gliadin T cell epitope selection by tissue transglutaminase in celiac disease. Role of enzyme specificity and pH influence on the transamidation versus deamidation process. J Biol Chem 277:34109-34116
32. Fukuda M, Satoh-Cruz M, Wen L, Crofts AJ, Sugino A, Washida H, Okita TW, Ogawa M, Kawagoe Y, Maeshima M, Kumamaru T (2011) The small GTPase Rab5a is essential for intracellular transport of proglutelin from the Golgi apparatus to the protein storage vacuole and endosomal membrane organization in developing rice endosperm. Plant Physiol 157:632-644
33. Fülöp V, Böcskei Z, Polgár L (1998) Prolyl oligopeptidase: an unusual beta-propeller domain regulates proteolysis. Cell 94:161-170
34. Fülöp V, Szeltner Z, Polgar L (2000) Catalysis of serine oligopeptidases is controlled by a gating filter mechanism. Embo Reports 1:277-281
35. Garner CP, Murray JA, Ding YC, Tien Z, van Heel DA, Neuhausen SL (2009) Replication of celiac disease UK genome-wide association study results in a US population. Hum Mol Genet 18:4219-4225
36. Gass J, Khosla C (2007) Prolyl endopeptidases. Cell Mol Life Sci 64:345-355
37. Gass J, Ehren J, Strohmeier G, Isaacs I, Khosla C (2005) Fermentation, purification, formulation, and pharmacological evaluation of a prolyl endopeptidase from Myxococcus xanthus: implications for celiac sprue therapy. Biotechnol Bioeng 92:674-684
38. Gass J, Vora H, Bethune MT, Gray GM, Khosla C (2006) Effect of barley endoprotease EP-B2 on gluten digestion in the intact rat. J Pharmacol Exp Ther 318:1178-1186
39. Gass J, Bethune MT, Siegel M, Spencer A, Khosla C (2007) Combination enzyme therapy for gastric digestion of dietary gluten in patients with celiac sprue. Gastroenterology 133:472-480
40. Gianfrani C, Siciliano RA, Facchiano AM, Camarca A, Mazzeo MF, Costantini S, Salvati VM, Maurano F, Mazzarella G, Iaquinto G, Bergamo P, Rossi M (2007) Transamidation of wheat flour inhibits the response to gliadin of intestinal T cells in celiac disease. Gastroenterology 133:780-789
41. Gil-Humanes J, Piston F, Hernando A, Alvarez JB, Shewry PR, Barro F (2008) Silencing of gamma-gliadins by RNA interference (RNAi) in bread wheat. J Cereal Sci 48:565-568
42. Gil-Humanes J, Piston F, Tollefsen S, Sollid LM, Barro F (2010) Effective shutdown in the expression of celiac disease-related wheat gliadin T-cell epitopes by RNA interference. Proc Nat Acad Sci USA 107:17023-17028
43. Gobbetti M, Rizzello CG, Di Cagno R, De Angelis M (2007) Sourdough lactobacilli and celiac disease. Food Microbiol 24:187-196
44. Gorrao SS, Hemerly JP, Lima AR, Melo RL, Szeltner Z, Polgar L, Juliano MA, Juliano L (2007) Fluorescence resonance energy transfer (FRET) peptides and cycloretro-inverso peptides derived from bradykinin as substrates and inhibitors of prolyl oligopeptidase. Peptides 28:2146-2154
45. Green P, Cellier C (2007) Celiac disease. N Engl J Med 357:1731- 1743
46. Gupta RB, Macritchie F (1994) Allelic variation at glutenin subunit and gliadin loci, Glu-1, Glu-3 and Gli-1 of common wheats 2. Biochemical basis of the allelic effects on dough properties. J Cereal Sci 19:19-29
47. Gupta PK, Mir RR, Mohan A, Kumar J (2008) Wheat genomics: present status and future prospects. Int J Plant Genomics 2008:896451
48. Haimila K, Einarsdottir E, de Kauwe A, Koskinen LL, Pan- Hammarstrom Q, Kaartinen T, Kurppa K, Ziberna F, Not T, Vatta S, Ventura A, Korponay-Szabo IR, Adany R, Pocsai Z, Szeles G, Dukes E, Kaukinen K, Maki M, Koskinen S, Partanen J, Hammarstrom L, Saavalainen P (2009) The shared CTLA4- ICOS risk locus in celiac disease, IgA deficiency and common variable immunodeficiency. Genes Immun 10:151-161
49. Halstensen TS, Scott H, Fausa O, Brandtzaeg P (1993) Gluten stimulation of celiac mucosa in-vitro induces activation (Cd25) of lamina propria Cd4(+) T-cells and macrophages but no cryptcell hyperplasia. Scand J Immunol 38:581-590
50. Hartmann G, Koehler P, Wieser H (2006) Rapid degradation of gliadin peptides toxic for coeliac disease patients by proteases from germinating cereals. J Cereal Sci 44:368-371
51. Hausch F, Santiago NA, Khosla C, Gray GM (2002a) Digestive resistance of immunodominant gliadin peptides: implications for enzyme therapy in celiac sprue. Gastroenterology 122:S1192
52. Hausch F, Shan L, Santiago NA, Gray GM, Khosla C (2002b) Intestinal digestive resistance of immunodominant gliadin peptides. Am J Physiol Gastrointest Liver Physiol 283:G996-G1003
53. Henderson KN, Reid HH, Borg NA, Broughton SE, Huyton T, Anderson RP, McCluskey J, Rossjohn J (2007) The production and crystallization of the human leukocyte antigen class II molecules HLA-DQ2 and HLA-DQ8 complexed with deamidated gliadin peptides implicated in coeliac disease. Acta Crystallogr Sect F Struct Biol Cryst Commun 63:1021-1025
54. Heyman M, Menard S (2009) Pathways of gliadin transport in celiac disease. Ann N Y Acad Sci 1165:274-278
55. Horvath H, Huang JT, Wong O, Kohl E, Okita T, Kannangara CG, von Wettstein D (2000) The production of recombinant proteins in transgenic barley grains. Proc Natl Acad Sci USA 97:1914-1919
56. Huibregtse IL, Marietta EV, Rashtak S, Koning F, Rottiers P, David CS, van Deventer SJ, Murray JA (2009) Induction of antigenspecific tolerance by oral administration of Lactococcus lactis delivered immunodominant DQ8-restricted gliadin peptide in sensitized nonobese diabetic Abo Dq8 transgenic mice. J Immunol 183:2390-2396
57. Hunt KA, Zhernakova A, Turner G, Heap GA, Franke L, Bruinenberg M, Romanos J, Dinesen LC, Ryan AW, Panesar D, Gwilliam R, Takeuchi F, McLaren WM, Holmes GK, Howdle PD, Walters JR, Sanders DS, Playford RJ, Trynka G, Mulder CJ, Mearin ML, Verbeek WH, Trimble V, Stevens FM, O'Morain C, Kennedy NP, Kelleher D, Pennington DJ, Strachan DP, McArdle WL, Mein CA, Wapenaar MC, Deloukas P, McGinnis R, McManus R, Wijmenga C, van Heel DA (2008) Newly identified genetic risk variants for celiac disease related to the immune response. Nat Genet 40:395-402
58. Jabri B, Sollid LM (2009) Tissue-mediated control of immunopathology in coeliac disease. Nat Rev Immunol 9:858-870
59. Johannsen B, Gjertsen H, Vartdal F, Buus S, Thorsby E, Lundin K, Sollid L (1996) Binding of peptides from the N-terminal region of α-gliadin to the celiac disease-associated HLA-DQ2 molecule assessed in biochemical and T cell assays. Clin Immunol Immunopathol 79:288-293
60. Juhasz T, Szeltner Z, Polgar L (2006) Properties of the prolyl oligopeptidase homologue from Pyrococcus furiosus. FEBS Lett 580:3493-3497
61. Juse U, van de Wal Y, Koning F, Sollid LM, Fleckenstein B (2010) Design of new high-affinity peptide ligands for human leukocyte antigen-DQ2 using a positional scanning peptide library. Hum Immunol 71:475-481
62. Kagnoff M, Harwood J, Bugawan T, Erlich H (1989) Structural analysis of the HLA-DR, -DQ, and -DP alleles on celiac diseaseassociated HLA-DR3 (DRw17) haplotype. Proc Natl Acad Sci USA 86:6274-6278
63. Keech CL, Dromey J, Chen Z, Anderson RP, McCluskey J (2009) Immune tolerance induced by peptide immunotherapy in an HLADQ2- dependent mouse model of gluten immunity. Gastroenterology 136:A355
64. Keil B (1992) Specificity of proteolysis. Springer- Verlag, 336p
65. Kim CY, Quarsten H, Bergseng E, Khosla C, Sollid LM (2004) Structural basis for HLA-DQ2-mediated presentation of gluten epitopes in celiac disease. Proc Natl Acad Sci USA 101:4175- 4179
66. Koskinen LL, Einarsdottir E, Dukes E, Heap GA, Dubois P, Korponay- Szabo IR, Kaukinen K, Kurppa K, Ziberna F, Vatta S, Not T, Ventura A, Sistonen P, Adany R, Pocsai Z, Szeles G, Maki M, Kere J, Wijmenga C, van Heel DA, Saavalainen P (2009) Association study of the IL18RAP locus in three European populations with coeliac disease. Hum Mol Genet 18:1148- 1155
67. Kumamaru T, Uemura Y, Inoue Y, Takemoto Y, Siddiqui SU, Ogawa M, Hara-Nishimura I, Satoh H (2010) Vacuolar processing enzyme plays an essential role in the crystalline structure of glutelin in rice seed. Plant Cell Physiol 51:38-46
68. Lafiandra D, Kasarda D (1985) One and two dimensional (two pH) polyacrilamide gel electrophoresis in a single gel: separation of wheat proteins. Cereal Chem 62:314-319
69. Lammers KM, Lu R, Brownley J, Lu B, Gerard C, Thomas K, Rallabhandi P, Shea-Donohue T, Tamiz A, Alkan S, Netzel- Arnett S, Antalis T, Vogel SN, Fasano A (2008) Gliadin induces an increase in intestinal permeability and zonulin release by binding to the chemokine receptor CXCR3. Gastroenterology 135:194-204
70. Li M, Chen CA, Davies DR, Chiu TK (2010) Induced-fit mechanism for prolyl endopeptidase. J Biol Chem 285:21487-21495
71. Lorand L, Graham RM (2003) Transglutaminases: crosslinking enzymes with pleiotropic functions. Nat Rev Mol Cell Biol 4:140-156
72. Lundin K, Sollid L, Qvigstad E, Markussen G, Gjertsen H, Ek J, Thorsby E (1990) T lymphocyte recognition of a celiac disease associated cis- or trans-encoded HLA-DQ a/b-heterodimer. J Immunol 145:136-139
73. Lundin KEA, Scott H, Hansen T, Paulsen G, Halstensen TS, Fausa O, Thorsby E, Sollid LM (1993) Gliadin-specitlc, HLA-DQ (crl.*0501, I.*0201) restricted T cells isolated from the small intestinal mucosa of celiac disease patients. J Exp Med 178:187-196
74. Lundin KEA, Gjertsen HA, Scott H, Sollid LM, Thorsby E (1994) Function of Dq2 and Dq8 as HLA susceptibility molecules in celiac-disease. Hum Immunol 41:24-27
75. Mahfouz MM, Li L, Shamimuzzaman M,Wibowo A, Fang X, Zhu J-K (2011) De novo-engineered transcription activator-like effector (TALE) hybrid nuclease with novel DNA binding specificity creates double-strand breaks. Proc Natl Acad Sci USA 108:2623-2628
76. Mamone G, Ferranti P, Rossi M, Roepstorff P, Fierro O, Malorni A, Addeo F (2007) Identification of a peptide from alpha-gliadin resistant to digestive enzymes: implications for celiac disease. J Chromatogr B Analyt Technol Biomed Life Sci 855:236-241
77. Marti T, Molberg O, Li Q, Gray GM, Khosla C, Sollid LM (2005) Prolyl endopeptidase-mediated destruction of T cell epitopes in whole gluten: chemical and immunological characterization. J Pharmacol Exp Ther 312:19-26
78. Matysiak-Budnik T, Candalh C, Cellier C, Dugave C, Namane A, Vidal-Martinez T, Cerf-Bensussan N, Heyman M (2005) Limited efficiency of prolyl-endopeptidase in the detoxification of gliadin peptides in celiac disease. Gastroenterology 129:786-796
79. Matysiak-Budnik T, Malamut G, de Serre NPM, Grosdidier E, Seguier S, Brousse N, Caillat-Zucman S, Cerf-Bensussan N, Schmitz J, Cellier C (2007) Long-term follow-up of 61 coeliac patients diagnosed in childhood: evolution toward latency is possible on a normal diet. Gut 56:1379-1386
80. Matysiak-Budnik T, Moura IC, Arcos-Fajardo M, Lebreton C, Menard S, Candalh C, Ben-Khalifa K, Dugave C, Tamouza H, van Niel G, Bouhnik Y, Lamarque D, Chaussade S, Malamut G, Cellier C, Cerf-Bensussan N, Monteiro RC, Heyman M (2008) Secretory IgA mediates retrotranscytosis of intact gliadin peptides via the transferrin receptor in celiac disease. J Exp Med 205:143-154
81. Maurano F, Siciliano RA, De Giulio B, Luongo D, Mazzeo MF, Troncone R, Auricchio S, Rossi M (2001) Intranasal administration of one alpha gliadin can downregulate the immune response to whole gliadin in mice. Scand J Immunol 53:290-295
82. Mention J-J, Ben Ahmed M, Bègue B, Barbe U, Verkarre V, Asnafi V, Colombel J-F, Cugnenc P-H, Ruemmele FM, McIntyre E, Brousse N, Cellier C, Cerf-Bensussan N (2003) Interleukin 15: a key to disrupted intraepithelial lymphocyte homeostasis and lymphomagenesis in celiac disease. Gastroenterology 125:730-745
83. Miletic ID, Sattelymiller EA, Schiffman SS, Miletic VD, Stankovic IM (1995) Identification of gliadin presence in pharmaceutical products - appropriateness polyclonal antibodies for detection of gliadin and other toxic prolamins. J Pediatr Gastroenterol Nutr 21:482-483
84. Mitea C, Havenaar R, Drijfhout JW, Edens L, Dekking L, Koning F (2008) Efficient degradation of gluten by a prolyl endoprotease in a gastrointestinal model: implications for coeliac disease. Gut 57:25-32
85. Molberg O, McAdam SN, Korner R, Quarsten H, Kristiansen C, Madsen L, Fugger L, Scott H, Noren O, Roepstorff P, Lundin KEA, Sjostrom H, Sollid LM (1998) Tissue transglutaminase selectively modifies gliadin peptides that are recognized by gut-derived T cells in celiac disease. Nat Med 4:713-717
86. Molberg O, McAdam S, Lundin KEA, Kristiansen C, Arentz-Hansen H, Kett K, Sollid LM (2001) T cells from celiac disease lesion recognize gliadin epitopes deamidated in situ by endogenous tissue transglutaminase. Eur J Immunol 31:1317-1323
87. Molberg O, Uhlen AK, Jensen T, Flaete NS, Fleckenstein B, Arentz- Hansen H, Raki M, Lundin KEA, Sollid LM (2005) Mapping of gluten T-cell epitopes in the bread wheat ancestors: implications for Celiac disease. Gastroenterology 128:393-401
88. Moriyama A, Nakanishi M, Sasaki M (1988a) Porcine muscle prolyl endopeptidase and its endogenous substrates. J Biochem 104:112-117
89. Moriyama A, Nakanishi M, Takenaka O, Sasaki M (1988b) Porcine muscle prolyl endopeptidase - limited proteolysis of tryptic peptides from hemoglobin beta-chains at prolyl and alanyl bonds. Biochim Biophys Acta 956:151-155
90. Nilsen EM, Lundin KEA, Krajci P, Scott H, Sollid LM, Brandtzaeg P (1995) Gluten specific, HLA-DQ restricted T-cells from celiac mucosa produce cytokines with Thl or Th0 profile dominated by interferon γ. Gut 37:766-776
91. Nolte WM, Tagore DM, Lane WS, Saghatelian A (2009) Peptidomics of prolyl endopeptidase in the central nervous system. Biochemistry 48:11971-11981
92. Norris JM, Barriga K, Hoffenberg EJ, Taki I, Miao DM, Haas JE, Emery LM, Sokol RJ, Erlich HA, Eisenbarth GS, Rewers M (2005a) Risk of celiac disease autoimmunity and timing of gluten introduction in the diet of infants at increased risk of disease. JAMA 293:2343-2351
93. Norris JM, Poole JA, Barriga K, Hoffenberg E, Rewers M (2005b) Timing of cereal introduction in the infant diet and risk of islet autoimmunity, celiac disease autoimmunity and wheat allergy: a clue to a common etiology? Diabetologia 48:303
94. Noula C, Kokotos G, Barth T, Tzougraki C (1997) New fluorogenic substrates for the study of secondary specificity of prolyl oligopeptidase. J Pept Res 49:46-51
95. Paterson BM, Lammers KM, Arrieta MC, Fasano A, Meddings JB (2007) The safety, tolerance, pharmacokinetic and pharmacodynamic effects of single doses of AT-1001 in coeliac disease subjects: a proof of concept study. Aliment Pharmacol Ther 26:757-766
96. Pavone P, Nicolini E, Taibi R, Ruggieri M (2007) Rotavirus and celiac disease. Am J Gastroenterol 102:1831
97. Payne PI, Holt LM, Law CN (1981) Structural and genetic-studies on the high-molecular-weight subunits of wheat glutenin 1. Allelic variation in subunits amongst varieties of wheat (Triticum aestivum). Theor Appl Genet 60:229-236
98. Payne PI, Nightingale MA, Krattiger AF, Holt LM (1987) The relationship between HMW glutenin subunit composition and the bread-making quality of British-grown wheat-varieties. J Sci Food Agric 40:51-65
99. Pinier M, Verdu EF, Nasser-Eddine M, David CS, Vezina A, Rivard N, Leroux JC (2009) Polymeric binders suppress gliadin-induced toxicity in the intestinal epithelium. Gastroenterology 136:288- 298
100. Piper JL, Gray GM, Khosla C (2002) High selectivity of human tissue transglutaminase for immunoactive gliadin peptides: implications for celiac sprue. Biochemistry 41:386-393
101. Piper JL, Gray GM, Khosla C (2004) Effect of prolyl endopeptidase on digestive-resistant gliadin peptides in vivo. J Pharmacol Exp Ther 311:213-219
102. Polgar L (1992) Unusual secondary specificity of prolyl oligopeptidase and the different reactivities of its 2 forms toward charged substrates. Biochemistry 31:7729-7735
103. Polgar L (2002a) The prolyl oligopeptidase family. Cell Mol Life Sci 59:349-362
104. Polgar L (2002b) Structure-function of prolyl oligopeptidase and its role in neurological disorders. Curr Med Chem Cent Nerv Syst Agents 2:251-257
105. Polgar L, Patthy A (1992) Cleavage of the Lys196-Ser197 bond of prolyl pligopeptidase - enhanced catalytic activity for one of the 2 active enzyme forms. Biochemistry 31:10769-10773
106. Potapov V, Cohen M, Schreiber G (2009) Assessing computational methods for predicting protein stability upon mutation: good on average but not in the details. Protein Eng Des Sel 22:553-560
107. Pyle GG, Paaso B, Anderson BE, Allen DD, Marti T, Li Q, Siegel M, Khosla C, Gray GM (2005) Effect of pretreatment of food gluten with prolyl endopeptidase on gluten-induced malabsorption in celiac sprue. Clin Gastroenterol Hepatol 3:687-694
108. Qiao SW, Bergseng E, Molberg O, Xia J, Fleckenstein B, Khosla C, Sollid LM (2004) Antigen presentation to celiac lesion-derived T cells of a 33-mer gliadin peptide naturally formed by gastrointestinal digestion. J Immunol 173:1757-1762
109. 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 175:254-261
110. Rennex D, Hemmings BA, Hofsteenge J, Stone SR (1991) cDNA cloning of Porcine brain prolyl endopeptidase and identification of the active-site seryl residue. Biochemistry 30:2195-2203
111. Rizzello CG, De Angelis M, Di Cagno R, Camarca A, Silano M, Losito A, De Vincenzi M, De Bari MD, Palmisano F, Maurano F, Gianfrani C, Gobbetti M (2007) Highly efficient gluten degradation by lactobacilli and fungal proteases during food processing: new perspectives for celiac disease. Appl Environ Microbiol 73:4499-4507
112. Romanos J, Barisani D, Trynka G, Zhernakova A, Bardella MT, Wijmenga C (2009) Six new coeliac disease loci replicated in an Italian population confirm association with coeliac disease. J Med Genet 46:60-63
113. Rossi M, Maurano F, Caputo N, Auricchio S, Sette A, Capparelli R, Troncone R (1999) Intravenous or intranasal administration of gliadin is able to down-regulate the specific immune response in mice. Scand J Immunol 50:177-182
114. Rustgi S, Brew-Appiah RAT, Wen N, Ankrah N, Wen S, Gemini R, Brueggeman R, Kannangara CG, Claar M, Langen G, Kogel K-H, Pang J, Liu B, von Wettstein D (2010) Epigenetics for the elimination of celiac-causing epitopes from wheat grains. Paper presented at the Joint Annual Meeting American Society of Plant Biologists and Canadian Society of Plant Physiology 2010, Montreal, Canada
115. Sanger GJ (2007) Endocannabinoids and the gastrointestinal tract: what are the key questions? Br J Pharmacol 152:663-670
116. Schumann M, Richter JF, Wedell I, Moos V, Zimmermann-Kordmann M, Schneider T, Daum S, Zeitz M, Fromm M, Schulzke JD (2008) Mechanisms of epithelial translocation of the alpha(2)- gliadin-33mer in coeliac sprue. Gut 57:747-754
117. Schuppan D (2000) Current concepts of celiac disease pathogenesis. Gastroenterology 119:234-242
118. Schuppan D, Junker Y, Barisani D (2009) Celiac disease: from pathogenesis to novel therapies. Gastroenterology 137:1912-1933
119. Senger S, Luongo D, Maurano F, Mazzeo MF, Siciliano RA, Gianfrani C, David C, Troncone R, Auricchio S, Rossi M (2003) Intranasal administration of a recombinant alpha-gliadin down-regulates the immune response to wheat gliadin in DQ8 transgenic mice. Immunol Lett 88:127-134
120. Shan L, Molberg O, Parrot I, Hausch F, Filiz F, Gray GM, Sollid LM, Khosla C (2002) Structural basis for gluten intolerance in celiac sprue. Science 297:2275-2279
121. Shan L, Martin T, Sollid LM, Gray GM, Khosla C (2004) Comparative biochemical analysis of three bacterial prolyl endopeptidases: implications for coeliac sprue. Biochem J 383:311-318
122. Shan L, Mathews II, Khosla C (2005a) Structural and mechanistic analysis of two prolyl endopeptidases: role of interdomain dynamics in catalysis and specificity. Proc Natl Acad Sci USA 102:3599-3604
123. Shan L, Qiao SW, Arentz-Hansen H, Molberg O, Gray GM, Sollid LM, Khosla C (2005b) Identification and analysis of multivalent proteolytically resistant peptides from gluten: implications for celiac sprue. J Proteome Res 4:1732-1741
124. Siegel M, Bethune MT, Gass J, Ehren J, Xia J, Johannsen A, Stuge TB, Gray GM, Lee PP, Khosla C (2006) Rational design of combination enzyme therapy for celiac sprue. Chem Biol 13:649-658
125. Siegel M, Strnad P, Watts RE, Choi K, Jabri B, Adler G, Ornary B, Khosla C (2008) Extracellular transglutaminase 2 is catalytically inactive, but is transiently activated upon tissue injury in the small intestine. Gastroenterology 134:A151
126. Sollid LM (2002) Coeliac disease: dissecting a complex inflammatory disorder. Nat Rev Immunol 2:647-655
127. Sollid LM, Jabri B (2011) Celiac disease and transglutaminase 2: a model for posttranslational modification of antigens and HLA association in the pathogenesis of autoimmune disorders. Curr Opin Immunol 23:732-738
128. Sollid LM, Markussen G, Ek J, Gjerde H, Vartdal F, Thorsby E (1989) Evidence for a primary association of celiac-disease to a particular HLA-DQ alpha-beta heterodimer. J Exp Med 169:345-350
129. Spaenij-Dekking L, Kooy-Winkelaar Y, van Veelen P, Drijfhout JW, Jonker H, van Soest L, Smulders MJ, Bosch D, Gilissen LJ, Koning F (2005) Natural variation in toxicity of wheat: potential for selection of nontoxic varieties for celiac disease patients. Gastroenterology 129:797-806
130. Spurkland A, Celius EG, Knutsen I, Beiske A, Thorsby E, Vartdal F (1997) The HLA-DQ(alpha 1.*0102, beta 1.*0602) heterodimer may confer susceptibility to multiple sclerosis in the absence of the HLA-DR(alpha 1.*01, beta 1.*1501) heterodimer. Tissue Antigens 50:15-22
131. Stene LC, Honeyman MC, Hoffenberg EJ, Haas JE, Sokol RJ, Emery L, Taki I, Norris JM, Erlich HA, Eisenbarth GS, Rewers M (2006) Rotavirus infection frequency and risk of celiac disease autoimmunity in early childhood: a longitudinal study. Am J Gastroenterol 101:2333-2340
132. 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 disease. Am J Physiol Gastrointest Liver Physiol 291:G621-G629
133. Stepniak D, Wiesner M, de Ru AH, Moustakas AK, Drijfhout JW, Papadopoulos GK, van Veelen PA, Koning F (2008) Large-scale characterization of natural ligands explains the unique glutenbinding properties of HLA-DQ2. J Immunol 180:3268-3278
134. Stern M, Ciclitira PJ, van Eckert R, Feighery C, Janssen FW, Mendez E, Mothes T, Troncone R, Wieser H (2001) Analysis and clinical effects of gluten in coeliac disease. Eur J Gastroenterol Hepatol 13:741-747
135. Stone SR, Rennex D, Wikstrom P, Shaw E, Hofsteenge J (1991) Inactivation of prolyl endopeptidase by a peptidylchloromethane - kinetics of inactivation and identification of sites of modification. Biochem J 276:837-840
136. Szeltner Z, Rea D, Juhasz T, Renner V, Fulop V, Polgar L (2004) Concerted structural changes in the peptidase and the propeller domains of prolyl oligopeptidase are required for substrate binding. J Mol Biol 340:627-637
137. Tatham AS, Gilbert SM, Fido RJ, Shewry PR (2000) Extraction, separation, and purification of wheat gluten proteins and related proteins of barley, rye, and oats. Methods Mol Med 41:55-73
138. Tenorio-Laranga J, Valero ML, Mannisto PT, del Pino MS, Garcia- Horsman JA (2009) Combination of snap freezing, differential pH two-dimensional reverse-phase high-performance liquid chromatography, and iTRAQ technology for the peptidomic analysis of the effect of prolyl oligopeptidase inhibition in the rat brain. Anal Biochem 393:80-87
139. Tjon JM, van Bergen J,Koning F (2010) Celiac disease: howcomplicated can it get? Immunogenetics 62:641-651
140. Tollefsen S, Arentz-Hansen H, Fleckenstein B, Molberg O, Lundin KEA, Sollid LM (2006a) DQ2 and DQ8 celiac lesion T-cells recognize differentially deamidated variants of the same gluten peptide; potential relevance for diabetes type I. Tissue Antigens 67:O-01
141. Tollefsen S, Arentz-Hansen H, Fleckenstein B, Molberg O, Raki M, Kwok WW, Jung G, Lundin KEA, Sollid LM (2006b) HLA-DQ2 and-DQ8 signatures of gluten T cell epitopes in celiac disease. J Clin Invest 116:2226-2236
142. Tripathi A, Lammers KM, Goldblum S, Shea-Donohue T, Netzel- Arnett S, Buzza MS, Antalis TM, Vogel SN, Zhao A, Yang S, Arrietta MC, Meddings JB, Fasano A (2009) Identification of human zonulin, a physiological modulator of tight junctions, as prehaptoglobin-2. Proc Natl Acad Sci USA 106:16799-16804
143. Trynka G, Zhernakova A, Romanos J, Franke L, Hunt KA, Turner G, Bruinenberg M, Heap GA, Platteel M, Ryan AW, de Kovel C, Holmes GK, Howdle PD, Walters JR, Sanders DS, Mulder CJ, Mearin ML, Verbeek WH, Trimble V, Stevens FM, Kelleher D, Barisani D, Bardella MT, McManus R, van Heel DA, Wijmenga C (2009) Coeliac disease-associated risk variants in TNFAIP3 and REL implicate altered NF-kappaB signalling. Gut 58:1078-1083
144. Trynka G, Hunt KA, Bockett NA, Romanos J, Mistry V, Szperl A, Bakker SF, Bardella MT, Bhaw-Rosun L, Castillejo G, de la Concha EG, de Almeida RC, Dias K-RM, van Diemen CC, Dubois PCA, Duerr RH, Edkins S, Franke L, Fransen K, Gutierrez J, Heap GAR, Hrdlickova B, Hunt S, Plaza Izurieta L, Izzo V, Joosten LAB, Langford C, Mazzilli MC, Mein CA, Midah V, Mitrovic M, Mora B, Morelli M, Nutland S, Nunez C, Onengut-Gumuscu S, Pearce K, Platteel M, Polanco I, Potter S, Ribes-Koninckx C, Ricano-Ponce I, Rich SS, Rybak A, Luis Santiago J, Senapati S, Sood A, Szajewska H, Troncone R, Varade J, Wallace C, Wolters VM, Zhernakova A, Thelma BK, Cukrowska B, Urcelay E, Ramon Bilbao J, Mearin ML, Barisani D, Barrett JC, Plagnol V, Deloukas P, Wijmenga C, van Heel DA, Spanish Consortium Genetics C, Prevent CDSG, Wtccc (2011) Dense genotyping identifies and localizes multiple common and rare variant association signals in celiac disease. Nat Genet 43:1193-1201
145. Tye-Din JA, Stewart JA, Dromey JA, Beissbarth T, van Heel DA, Tatham A, Henderson K, Mannering SI, Gianfrani C, Jewell DP, Hill AVS, McCluskey J, Rossjohn J, Anderson RP (2010) Comprehensive, quantitative mapping of T cell epitopes in gluten in celiac disease. Sci Transl Med 2:41ra51
146. Vader W (2002) The gluten response in children with celiac disease is directed toward multiple gliadin and glutenin peptides. Gastroenterology 122:1729-1737
147. Vader LW, de Ru A, van der Wal Y, Kooy YMC, Benckhuijsen W, Mearin ML, Drijfhout JW, van Veelen P, Koning F (2002) Specificity of tissue transglutaminase explains cereal toxicity in celiac disease. J Exp Med 195:643-649
148. van Belzen MJ, Mulder CJ, Zhernakova A, Pearson PL, Houwen RH, Wijmenga C (2004) CTLA4 +49 A/G and CT60 polymorphisms in Dutch coeliac disease patients. Eur J Hum Genet 12:782-785
149. van de Wal Y, Kooy YMC, Drijfhout JW, Amons R, Koning F (1996) Peptide binding characteristics of the coeliac disease-associated DQ(alpha 1.*0501, beta 1.*0201) molecule. Immunogenetics 44:246-253
150. van de Wal Y, Kooy Y, Van Veelen P, Pena S, Mearin L, Molberg O, Lundin K, Sollid L, Mutis T, Benckhuijsen W, Drijfhout JW, Koning F (1998a) T cells from the small intestinal mucosa of coeliac disease patients recognize unique peptide sequence of gliadin. Gastroenterology 114:A1105
151. van de Wal Y, Kooy Y, van Veelen P, Pena S, Mearin L, Papadopoulos G, Koning F (1998b) Cutting edge: selective deamidation by tissue transglutaminase strongly enhances gliadin-specific T cell reactivity. J Immunol 161:1585-1588
152. van de Wouw M, Kik C, van Hintum T, van Treuren R, Visser B (2010a) Genetic erosion in crops: concept, research results and challenges. Plant Genet Resour Charact Util 8:1-15
153. van de Wouw M, van Hintum T, Kik C, van Treuren R, Visser B (2010b) Genetic diversity trends in twentieth century crop cultivars: a meta analysis. Theor Appl Genet 120:1241-1252
154. van den Broeck HC, van Herpen T, Schuit C, Salentijn EMJ, Dekking L, Bosch D, Hamer RJ, Smulders MJM, Gilissen L, van der Meer IM (2009) Removing celiac disease-related gluten proteins from bread wheat while retaining technological properties: a study with Chinese Spring deletion lines. BMC Plant Biol 9:41
155. van den Broeck HC, de Jong HC, Salentijn EM, Dekking L, Bosch D, Hamer RJ, Gilissen LJ, van der Meer IM, Smulders MJ (2010) Presence of celiac disease epitopes in modern and old hexaploid wheat varieties: wheat breeding may have contributed to increased prevalence of celiac disease. Theor Appl Genet 121:1527-1539
156. van Elzen R, Lambeir AM (2011) Structure and function relationship in prolyl oligopeptidase. CNS Neurol Disord Drug Targets 10:297-305
157. van Heel DA, Franke L, Hunt KA, Gwilliam R, Zhernakova A, Inouye M, Wapenaar MC, Barnardo MC, Bethel G, Holmes GK, Feighery C, Jewell D, Kelleher D, Kumar P, Travis S, Walters JR, Sanders DS, Howdle P, Swift J, Playford RJ, McLaren WM, Mearin ML, Mulder CJ, McManus R, McGinnis R, Cardon LR, Deloukas P, Wijmenga C (2007) A genome-wide association study for celiac disease identifies risk variants in the region harboring IL2 and IL21. Nat Genet 39:827-829
158. van Herpen T, Goryunova SV, van der Schoot J, Mitreva M, Salentijn E, Vorst O, Schenk MF, van Veelen PA, Koning F, van Soest LJM, Vosman B, Bosch D, Hamer RJ, Gilissen L, Smulders MJM (2006) Alpha-gliadin genes from the A, B, and D genomes of wheat contain different sets of celiac disease epitopes. BMC Genomics 7:1
159. van Herpen TWJM, Riley M, Sparks C, Jones HD, Gritsch C, Dekking EH, Hamer RJ, Bosch D, Salentijn EMJ, Smulders MJM, Shewry PR, Gilissen LJWJ (2008) Detailed analysis of the expression of an alpha-gliadin promoter and the deposition of alpha-gliadin protein during wheat grain development. Ann Bot 102:331-342
160. Venalainen JI, Juvonen RO, Mannisto PT (2004) Evolutionary relationships of the prolyl oligopeptidase family enzymes. Eur J Biochem 271:2705-2715
161. Vilppula A, Kaukinen K, Luostarinen L, Krekela I, Patrikainen H, Valve R, Maki M, Collin P (2009) Increasing prevalence and high incidence of celiac disease in elderly people: a population-based study. BMC Gastroenterol 9:49
162. Vincentini O, Maialetti F, Gazza L, Silano M, Dessi M, De Vincenzi M, Pogna NE (2007) Environmental factors of celiac disease: cytotoxicity of hulled wheat species Triticum monococcum. T. turgidum ssp. dicoccum and T. aestivum ssp. spelta. J Gastroenterol Hepatol 22:1816-1822
163. von Wettstein D (2007) From analysis of mutants to genetic engineering. Annu Rev Plant Biol 58:1-19
164. von Wettstein D, Mikhaylenko G, Froseth JA, Kannangara CG (2000) Improved barley broiler feed with transgenic malt containing heat-stable (1,3-1,4)-beta-glucanase. Proc Natl Acad Sci USA 97:13512-13517
165. von Wettstein D, Warner J, Kannangara CG (2003) Supplements of transgenic malt or grain containing (1,3-1,4)-beta-glucanase increase the nutritive value of barley-based broiler diets to that of maize. Br Poult Sci 44:438-449
166. Wen S, Kannangara G, Rustgi S, Claar M, Langen G, Pang J, von Wettstein D Epigenetic Regulation of Wheat Gliadin and Low Molecular Weight Glutenins. In: Biologists ASoP (ed) Joint Annual Meeting American Society of Plant Biologists and Canadian Society of Plant Physiologists, Montreal, Canada, 2010. American Society of Plant Biologists
167. Wieser H (1996) Relation between gliadin structure and coeliac toxicity. Acta Paediatrica 85:3-9
168. Wieser H, Koehler P, Folck A, Becker D (2006) Characterization of wheat with strongly reduced α-gliadin content. In: Lookhart GL, Ng PKW (eds) Gluten proteins. AACC, St Paul
169. Wolters VM,Wijmenga C (2008) Genetic background of celiac disease and its clinical implications. Am J Gastroenterol 103:190-195
170. Wolters VM, Verbeek WHM, Zhernakova A, Onland-Moret C, Schreurs MWJ, Monsuur AJ, Verduijn W, Wijmenga C, Mulder CJJ (2008) The MY09B gene is a strong risk factor for the development of refractory coeliac disease. Clin Gastroenterol Hepatol 5:1399-1405
171. Yokoyama K, Nio N, Kikuchi Y (2004) Properties and applications of microbial transglutaminase. Appl Microbiol Biotechnol 64:447-454