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Applied and Environmental Microbiology, February 2004, p. 1088-1096, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1088-1096.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Sourdough Bread Made from Wheat and Nontoxic Flours and Started with Selected Lactobacilli Is Tolerated in Celiac Sprue Patients

Raffaella Di Cagno,1,{dagger} Maria De Angelis,2,{dagger} Salvatore Auricchio,3 Luigi Greco,3 Charmaine Clarke,4 Massimo De Vincenzi,5 Claudio Giovannini,5 Massimo D'Archivio,5 Francesca Landolfo,3 Giampaolo Parrilli,3 Fabio Minervini,1 Elke Arendt,4 and Marco Gobbetti1*

Department of Plant Protection and Applied Microbiology, University of Bari, 70126 Bari,1 Institute of Sciences of Food Production, CNR, 70100 Bari,2 European Laboratory for Food Induced Disease (ELFID), Department of Pediatrics and Gastroenterology, University of Naples Federico II, 80131 Naples,3 Istituto Superiore di Sanità, Laboratorio di Metabolismo e Biochimica Patologica, I-00161 Rome, Italy,5 Department of Food and Nutritional Sciences, University College Cork, Cork, Ireland4

Received 30 May 2003/ Accepted 10 October 2003

This work was aimed at producing a sourdough bread that is tolerated by celiac sprue (CS) patients. Selected sourdough lactobacilli had specialized peptidases capable of hydrolyzing Pro-rich peptides, including the 33-mer peptide, the most potent inducer of gut-derived human T-cell lines in CS patients. This epitope, the most important in CS, was hydrolyzed completely after treatment with cells and their cytoplasmic extracts (CE). A sourdough made from a mixture of wheat (30%) and nontoxic oat, millet, and buckwheat flours was started with lactobacilli. After 24 h of fermentation, wheat gliadins and low-molecular-mass, alcohol-soluble polypeptides were hydrolyzed almost totally. Proteins were extracted from sourdough and used to produce a peptic-tryptic digest for in vitro agglutination tests on K 562(S) subclone cells of human origin. The minimal agglutinating activity was ca. 250 times higher than that of doughs chemically acidified or started with baker's yeast. Two types of bread, containing ca. 2 g of gluten, were produced with baker's yeast or lactobacilli and CE and used for an in vivo double-blind acute challenge of CS patients. Thirteen of the 17 patients showed a marked alteration of intestinal permeability after ingestion of baker's yeast bread. When fed the sourdough bread, the same 13 patients had values for excreted rhamnose and lactulose that did not differ significantly from the baseline values. The other 4 of the 17 CS patients did not respond to gluten after ingesting the baker's yeast or sourdough bread. These results showed that a bread biotechnology that uses selected lactobacilli, nontoxic flours, and a long fermentation time is a novel tool for decreasing the level of gluten intolerance in humans.

* Corresponding author. Mailing address: Dipartimento di Protezione delle Piante e Microbiologia Applicata, Facoltà di Agraria di Bari, Via G. Amendola 165/a, 70126 Bari, Italy. Phone: 39 080 5442949. Fax: 39 080 5442911. E-mail: gobbetti{at}agr.uniba.it.

{dagger} R. Di Cagno and M. De Angelis contributed equally to this work.

Applied and Environmental Microbiology, February 2004, p. 1088-1096, Vol. 70, No. 2
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.2.1088-1096.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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