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Biochemical modifications of gliadins induced by microbial transglutaminase on wheat flour.
Biochim Biophys Acta. 2013 Nov; 1830(11):5166-74.BB

Abstract

BACKGROUND

Celiac disease (CD) is an immune-mediated disorder caused by the ingestion of wheat gluten. A lifelong, gluten-free diet is required to normalize the intestinal mucosa. We previously found that transamidation by microbial transglutaminase (mTGase) suppressed the gliadin-specific immune response in intestinal T-cell lines from CD patients and in models of gluten sensitivity.

METHODS

SDS-PAGE, Western blot, ELISA, tissue transglutaminase (tTGase) assay and nano-HPLC-ESI-MS/MS experiments were used to analyze prolamins isolated from treated wheat flour.

RESULTS

Gliadin and glutenin yields decreased to 7.6±0.5% and 7.5±0.3%, respectively, after a two-step transamidation reaction that produced a water-soluble protein fraction (spf). SDS-PAGE, Western blot and ELISA analyses confirmed the loss of immune cross-reactivity with anti-native gliadin antibodies in residual transamidated gliadins (K-gliadins) and spf as well as the occurrence of neo-epitopes. Nano-HPLC-ESI-MS/MS experiments identified some native and transamidated forms of celiacogenic peptides including p31-49 and confirmed that mTGase had similar stereo-specificity of tTGase. Those peptides resulted to be 100% and 57% modified in spf and K-gliadins, respectively. In particular, following transamidation p31-49 lost its ability to increase tTGase activity in Caco-2 cells. Finally, bread manufactured with transamidated flour had only minor changes in baking characteristics.

CONCLUSIONS

The two-step transamidation reaction modified the analyzed gliadin peptides, which are known to trigger CD, without influencing main technological properties.

GENERAL SIGNIFICANCE

Our data shed further light on a detoxification strategy alternative to the gluten free diet and may have important implications for the management of CD patients.

Authors+Show Affiliations

Institute of Food Sciences, CNR, Avellino, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23891939

Citation

Mazzeo, Maria F., et al. "Biochemical Modifications of Gliadins Induced By Microbial Transglutaminase On Wheat Flour." Biochimica Et Biophysica Acta, vol. 1830, no. 11, 2013, pp. 5166-74.
Mazzeo MF, Bonavita R, Maurano F, et al. Biochemical modifications of gliadins induced by microbial transglutaminase on wheat flour. Biochim Biophys Acta. 2013;1830(11):5166-74.
Mazzeo, M. F., Bonavita, R., Maurano, F., Bergamo, P., Siciliano, R. A., & Rossi, M. (2013). Biochemical modifications of gliadins induced by microbial transglutaminase on wheat flour. Biochimica Et Biophysica Acta, 1830(11), 5166-74. https://doi.org/10.1016/j.bbagen.2013.07.021
Mazzeo MF, et al. Biochemical Modifications of Gliadins Induced By Microbial Transglutaminase On Wheat Flour. Biochim Biophys Acta. 2013;1830(11):5166-74. PubMed PMID: 23891939.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Biochemical modifications of gliadins induced by microbial transglutaminase on wheat flour. AU - Mazzeo,Maria F, AU - Bonavita,Roberta, AU - Maurano,Francesco, AU - Bergamo,Paolo, AU - Siciliano,Rosa A, AU - Rossi,Mauro, Y1 - 2013/07/25/ PY - 2013/02/17/received PY - 2013/06/24/revised PY - 2013/07/18/accepted PY - 2013/7/30/entrez PY - 2013/7/31/pubmed PY - 2014/3/22/medline KW - CD KW - Celiac disease KW - GFD KW - Gliadin KW - K-C(2)H(5) KW - K-gliadins KW - Microbial transglutaminase KW - PEPs KW - Transamidation KW - gluten-free diet KW - insoluble transamidated gliadin KW - lysine ethyl ester KW - mTGase KW - microbial transglutaminase KW - nano-HPLC–ESI-MS/MS KW - prolyl endopeptidases KW - spf KW - tTGase KW - tandem mass spectrometry coupled with nano-reverse phase liquid chromatography KW - tissue transglutaminase KW - water-soluble protein fraction SP - 5166 EP - 74 JF - Biochimica et biophysica acta JO - Biochim Biophys Acta VL - 1830 IS - 11 N2 - BACKGROUND: Celiac disease (CD) is an immune-mediated disorder caused by the ingestion of wheat gluten. A lifelong, gluten-free diet is required to normalize the intestinal mucosa. We previously found that transamidation by microbial transglutaminase (mTGase) suppressed the gliadin-specific immune response in intestinal T-cell lines from CD patients and in models of gluten sensitivity. METHODS: SDS-PAGE, Western blot, ELISA, tissue transglutaminase (tTGase) assay and nano-HPLC-ESI-MS/MS experiments were used to analyze prolamins isolated from treated wheat flour. RESULTS: Gliadin and glutenin yields decreased to 7.6±0.5% and 7.5±0.3%, respectively, after a two-step transamidation reaction that produced a water-soluble protein fraction (spf). SDS-PAGE, Western blot and ELISA analyses confirmed the loss of immune cross-reactivity with anti-native gliadin antibodies in residual transamidated gliadins (K-gliadins) and spf as well as the occurrence of neo-epitopes. Nano-HPLC-ESI-MS/MS experiments identified some native and transamidated forms of celiacogenic peptides including p31-49 and confirmed that mTGase had similar stereo-specificity of tTGase. Those peptides resulted to be 100% and 57% modified in spf and K-gliadins, respectively. In particular, following transamidation p31-49 lost its ability to increase tTGase activity in Caco-2 cells. Finally, bread manufactured with transamidated flour had only minor changes in baking characteristics. CONCLUSIONS: The two-step transamidation reaction modified the analyzed gliadin peptides, which are known to trigger CD, without influencing main technological properties. GENERAL SIGNIFICANCE: Our data shed further light on a detoxification strategy alternative to the gluten free diet and may have important implications for the management of CD patients. SN - 0006-3002 UR - https://www.unboundmedicine.com/medline/citation/23891939/Biochemical_modifications_of_gliadins_induced_by_microbial_transglutaminase_on_wheat_flour_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0304-4165(13)00324-3 DB - PRIME DP - Unbound Medicine ER -