cristina m. rosell (iata-csic) - crosell@iata.csic.es - @iata.csic.escristina m. rosell (iata-csic)...

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Cristina M. Rosell (IATA-CSIC) - crosell@iata.csic.es - www.iata.csic.es

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Prof. Cristina M. Rosell

Enzymes in gluten-free bread making

Enzymes in gluten-free bread making by Cristina M. Rosell is licensed under a Creative Commons Reconocimiento-CompartirIgual 4.0 Internacional License.

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OUTLINE:

1. Role of enzymes in bakery

2. Gluten free products

3. Enzymes with structuring action

4. Enzymes with degrading activity

5. Enzymes for nutritionally improving gluten free breads

6. Concluding remarks

7. Acknowledgments

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Enzymes baking

industries

Starch-degrading enzymes

NSP degrading enzymes

Protein enzymes

Lipid enzymes

Other enzymes

α-amylase β-amylase

Glucoamylase

Debranching enzymes:

Isoamylase Pullulanase

Transferases: Amylomaltase

CGTase

Most commonly used

Less commonly used

Pentosanases: Hemicellulase

Xylanase

Proteases: Exopeptidase

Endopeptidases

Transglutaminase

Asparaginase

Phytase Glucose oxidase

Lipoxygenase Laccase

Lipases Phospholipase A

Rosell & Dura (2014). Enzymes in baking industries. In: Enzymes in Food and Beverage. Ed E. Chandrasekeran. CRC Press

1. Role of enzymes in bakery

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Enzymes Dough Bread Staling α-amylases Improve fermentation Improve volume and

crumb texture Anti-staling effect

CGT-ases Improve volume Decrease firmness

Slower staling kinetics

Pentosanases Improve handling Improve volume and crumb texture

Hemicellulases Reduce processing time Increase volume Xylanases Increase gluten strength

Improve handling and fermentative characteristics

Increase shelf-life Reduce bread-staling Increase shelf life

Proteases Easy mixing May compromise gas -holding ability

Tyrosinase

Increase hardness Decrease extensibility

Uneven crumbs and large pore size Soft texture and increase volume

Transglutaminase Decrease water absorption Increase stability Favors tiny gas cells

Increase volume and improve structure

Lipases Improve rheological properties Dough strengthener

Improve crumb structure Flavor enhancement Increase specific volume

Antistaling effect when combined with α-amylase

Phospholipase A Improve handling and suppress stickiness

Increase volume

Phytase Shorten fermentation Increase specific volume and soften crumbs Improve nutritional values

Glucose oxidase Dough strengthener Improve quality Lipoxygenase Improve visco-elasticity

Bleaching agent

Laccase Improve stabilization and machinability Dough strengthener

Soften crumb

Rosell & Dura (2014). Enzymes in baking industries. In: Enzymes in Food and Beverage. Ed E. Chandrasekeran. CRC Press

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Gluten forms a viscoelastic matrix necessary for holding the

CO2 released during the fermentation

1st Challenge: Design gluten free products

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Building an internal network:• Hydrocolloids • Proteins • Enzymes: GO, Laccase, PFO, TG

Rosell (2009). Enzymatic manipulation of gluten-free bread. In: Gluten-free Food Science and Technology. Ed E. Gallagher.

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β-D-glucose + O2 Gluconic Acid + H2O2

Mechanism of GO

Mechanism of TG

Acyl – transfer reaction

Crosslinking reaction

Deamidation

Rosell (2009). Enzymatic manipulation of gluten-free bread. In: Gluten-free Food Science and Technology. Ed E. Gallagher.

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Enzyme concentration (%,w/w)

0,00 0,01 0,02 0,030,00 0,01 0,02 0,030,00 0,01 0,02 0,030,00 0,01 0,02 0,03

0 % HPMC 2 % HPMC

Enzyme concentration (%,w/w)

0,00 0,01 0,02 0,030,00 0,01 0,02 0,030,00 0,01 0,02 0,030,00 0,01 0,02 0,03C

0 % HPMC 2 % HPMC

Glucose Oxidase // Rice bread

Gujral and Rosell (2004) Food Res Technol 37, 75-81.

Rosell (2009). Enzymatic manipulation of gluten-free bread. In: Gluten-free Food Science and Technology. Ed E. Gallagher

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Glucose Oxidase // Corn bread

Control GO 0.01 GO 0.020

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Transglutaminase // Rice bread

Gujral, Haros, Rosell (2004) J Cereal Sci 39, 225-230

Enzyme concentration (%, w/w)

0,0 0,5 1,0 1,5

Enzyme concentration (%, w/w)

0,0 0,5 1,0 1,50,0 0,5 1,0 1,5

0% HPMC2% HPMC

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Proteins: pea, soybean, albumen, casein Viscoelastic parameters

Level G΄ (Pa) G΄΄ (Pa) Tan

Overall mean 5372 663 0,176

Protein 0 6460b 719b 0,112a

1 8992c 1211c 0,145a

2 10902d 1273c 0,118a

3 382a 77a 0,205b

4 126a 33a 0,298c

TG 0 4412a 611a 0,168a

1 6333b 714a 0,183a

1 Values followed by different letters in the same column within each factor are significantly 1 different (p<0.05). 2 Protein levels: 0 (without protein isolate), 1 (pea), 2 (soybean), 3 (egg albumen), 4 (whey 3 protein). 4 TG levels: 0 (without TG), 1 (1% TG). 5

Marco and Rosell (2008) J Food Eng 84, 132-139

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Marco, Pérez, León, Rosell (2008) Cereal Chem 85, 59-64. Marco, Rosell (2008) Eur Food Research Technol. 227, 1205-1213

Rice-Soybean

+TGTime (min)

0 10 20 30 40 50

SP 25%-110

SP 25%-95

SP 13%-110

SP 13%-95

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Storck et al. (2013). LWT - Food Science and Technology. 53/ 1 (2013) 346–354.

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PROTEASE (Neutrase 1.5 MG, Novozymes; PR)

buckwheat (BW)sorghum (SG)corn (CR)

Renzetti and Arendt. Journal of Cereal Science 50, (2009): 22-28

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4. Enzymes with degrading activityPROTEASES: - protease A and M from Aspergillus oryzae- protease P from Aspergillus melleus- thermoase PC10F from Bacillus stearothermophilus) from Amano Enzyme Inc.

(Nagoya, Japan). - Thermolysin from Bacillus thermoproteolyticus rokko (Type X) from Sigma

Aldrich

Kawamura-Konishi et al. Journal of Cereal Science 58, no. 1 (2013): 45-50.

RICE BREAD

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4. Enzymes with degrading activityPROTEASES: - Alcalase (subtilisine) from Bacillus licheniformis- iZyme (whey protease) from Novozyme (Denmark)- Flavourzyme (aminopeptidase) from Novozyme

CORN BREAD

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PROTEASES: - iZyme (whey protease) from Novozyme

(Denmark)

CORN BREAD

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AMYLASES: - Fungamyl from Novozyme (Denmark)- Powerfresh (Dupont)

RICE BREAD

CONTROL

GFB AM

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FUTURE TRENDS

Enzymes for nutritionally improving gluten free breads

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SOURDOUGHS:

- Acidification

- production of exopolysaccharides,

- activation of enzymes like proteases, amylases and phytases

- production of antimicrobial substances like propionate

Detoxification of the gluten containing cereals

Reducing antinutrients

Improving quality

Moroni, Dal Bello, Arendt. Food microbiology. 2009;26(7):676-84.

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- GERMINATION:

- Activation of enzyme activities

- amaranth, buckwheat, maize, millet, rice, sorghum and

quinoa

Reduce anti-nutrients content

Reduce eGI

Omary, Fong, Rothschild, Finney. Cereal Chemistry. 2012; 89(1):1-14Cornejo & Rosell (2014) Unpublished results.

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PROLYL ENDOPEPTIDASES from different sources.

• Produce gluten-free foods from gluten-containing

raw materials

• Produce an oral therapy for CD

Wieser, Koehler. Journal of Aoac International. 2012, 95(2):356-63.

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FINAL REMARKS

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Enzymes are useful processing aids in gluten free bread making. Enzymes’ action is largely dependent on raw material, thus enzyme type and level must be optimized for each gluten free recipe.

Innovation: re-defining role of enzymes from “processing aids” to “healthy aids”.

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ACKNOWLEDGEMENTS:- Hardeep Singh Gujral (Guru Nanak University, India)

- Cristina Marco (IATA-CSIC)

- Mª Estela Matos (IATA-CSIC)

- Marcia Arocha Gularte (Universidad de Pelotas, Brasil)

- Fabiola Cornejo (ESPOL, Ecuador)

Funding sources:• Proyect UE. CRAFT Proyect (QLK1-2002-72162), 2003-2005• Proyect (AGL2005-05192-C04-01/ALI) by Spanish Ministry of Economy

and Sustainability and CSIC.• Award from Coelic Association of Madrid in 2009• Proyect (AGL2008- 00092 ALI , AGL2011-23802) by Spanish Ministry of

Economy and Sustainability and CSIC.

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For more information: Web site: http://www.iata.csic.ese-mail: crosell@iata.csic.esWebpage: http://www.iata.csic.es/mcristinamolinarosell/Research ID: http://www.researcherid.com/rid/F-4888-2010

Enzymes in gluten-free bread making by Cristina M. Rosell is licensed under a Creative Commons Reconocimiento-CompartirIgual 4.0 Internacional License.

cristina m. rosell (iata-csic) - crosell@iata.csic.es - @iata.csic.escristina m. rosell (iata-csic)...

rosell iatacsic

rosell dura

gluten free products

glutenfree bread

glutenfree food science

gallagher enzymes

structuring action enzymes

design gluten free products2

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