Dr John WilliamsThe Wheat Protein & Coeliac Consortium

http://commodityinstitute.com/latest-news/136-the-wheat-gluten-peptide-miracle-and-coeliac-beast

REFERENCE PAPER

Dr John Williams - Executive officer

Professor Rudi Appels – Chairman International Wheat Genome Sequencing Consortium

Dr Angela Juhasz, Murdoch University Agricultural Biotechnology Centre

Dr Geoffrey Annison – Health/Nutrition at the Australian Food & Grocery Council

Mark Laucke – Laucke Flour Mills

Ken Dods – Principal Food Scientist, WA ChemCentre

Dr Kristian Ronacher – food nutritionist (previously with CSIRO Food Division)

Professor Dean Diepeveen, Murdoch University Agricultural Biotechnology Centre

Associate Professor Chris Florides, Murdoch University Agricultural Biotechnology Centre

Hugo Kruik, Lesaffre and Australian Society of Bakers

Stephen Noonan, traditional bakers group; Dr Alistair Watson, wheat industry economist

CONSORTIUM TEAM:

True: Wheat has been feeding much of the world for 6-7000 years

- despite having offending gluten peptides

False: Wheat has been consumed for 6-7000 years

True – Wheaten products have been consumed but only after hydrolysis with water + enzyme activity

RAPID DOUGH NO-TIME DOUGH LESS TIME METHOD METHOD BUSINESS CULTURE

BAKERS’ OBJECTIVE: TO DECREASE TIME & COST

WHAT IS DIFFERENT TODAY:

1. LESS FERMENTATION TIME

PLANT MILLERS/BAKERS FOOD BREEDERS MANUFACTURERS

WHAT IS DIFFERENT TODAY:

2. ADDED GLUTEN (SINCE 1970) Source: Kasarda, USDA, 2013

IMPROVE RHEOLOGICAL PROPERTIES

INCREASE FOOD FUNCTIONALITY

ENHANCE RESISTANCE AGAINST PESTS & DISEASES

WHAT IS DIFFERENT TODAY:

3. MORE NON-FERMENTATED WHEAT FOODS

High incidence of coeliac disease in Mediterranean countries

Unleavened bread, cous-cous, spaghettis, pastas, biscuits, noodles, batters

Medical impact:• Chronic inflammatory coeliac disease or gluten intolerance• Coeliac disease is latent with triggering factors (eg inadequate microbia)• 40% of Caucasians have gene predisposition• No remission has been medically recorded - chronic• 3.5 million European and 2 million US people have coeliac disease• Diagnosis is costly and intrusive making un-diagnosis high • 22 percent of US consumers were following a gluten-free diet in 2014_________________________________________________

Market impact:• Flat wheat prices compared to other grains• Negative protein grade spreads – penalties for higher protein• Less value for protein wheat• Grain sorghum prices have been higher than milling wheat prices

WE SUSPECT THE REAL PROBLEM IS UNHYDROLYSED GLUTEN OVERLOAD CAUSING DIS-EASE

(> 20-50 grams of gluten per day)

Consortium Objective:

To make wheaten bread safer for consumersby hydrolysing the gluten

within normal commercial processesand within the confines of time and cost

SOLUTION LIMITS

Cannot simply remove wheat proteins a. complex gluten peptides - 16,000 fragmentsb. plant nutritional & diseases/pest benefitsc. visco-elastic-extensibility rheological propertiesd. issues with genetically-modified foods

Cannot simply remove added glutena. problems with dough variability & dough inconsistencyb. decreased dough elasticity when bran/grain are addedc. food manufacturing would have less functionalityd. many foods would be less saleable

Cannot simply remove non-fermented wheaten foodsa. there needs to be consumer choiceb. foods need to meet lifestyle demandsc. wheat products are nutritious

The International Wheat Genome Sequencing ConsortiumSource: Rudi Appels, 2015

RESEARCH BREAK-THROUGH1. THE BASIC GENOME (DNA HELIX) OF WHEAT

DNA molecules for optical mapping

Building a complete picture of the genome provides access to every gene coding for a protein

Source: Rudi Appels, 2015

RESEARCH BREAK-THROUGH2. ProPepper – database of cereal prolamin epitopes,

peptides, and proteins (Source: Angela Juhasz, 2015)

• 2,484 cereal proteins • 37,914 cereal peptides• 833 epitopes (antigens that trigger immune response)

• 94,924 protein-epitope connections• 119,069 peptide-epitope connections• 667,402 digestions by enzymes

MALDI-TOF EQUIPMENT IS USED TO IDENTIFY PROTEINS BASED ON MOLECULAR WEIGHT

28998 31359 33720 36081 38442Mass (m/z)

0

10

20

30

40

50

60

70

80

90

100

% Intensit

y

Voyager Spec #1=>AdvBC(256,0.1,0.1)=>NR(4.00)=>SM15[BP = 31590.1, 174]

31604

3139131220

31979

3209533768

31781

34000

3110030691

3058432307

3024835260

33474

30081

3909135057

34703 35560

3857236938

29137.0 32159.8 35182.6 38205.4Mass (m/z)

0

10

20

30

40

50

60

70

80

90

% Intensit

y

3206139142

31380

35272

3404531866

32291

30571

3124033485

337993070739352

30246

3545634242

3508030059

399236053 36979

Normal bread from shorter fermentation

Bread from longer fermentation

Source: Rudi Appels, 2015

- Then determine amino acid sequences through chromatography

- Then identify linkages to the coeliac epitopes

- Then have proof of concept throughhuman clinical trials

1. Test human reaction to different fermentation methods

2. Discover enzymes to denature offending peptides

3. Develop a commercial rapid test for peptide denaturing

4. Promote more appropriate food production techniques

5. Create the research basis for a sustainable health claim

RESEARCH DIRECTION

PROOF OF CONCEPTANALYTICS ZERO UNHYDROLYSED GLUTEN

1. This is an industry problem

2. There are serious limits to possible solutions

3. Need to test peptides-epitopes before & after using different fermentation methods

4. Need to identify the best fermentation-hydrolysis technique within commercial confines

5. Need to obtain proof of concept through human clinical trials

AIMS

A. To develop appropriate industry guidelines, protocols, and labelling

B. To improve dough fermentation effectiveness

C. To decrease effective dough fermentation time

D. To find a simple cheap quick gluten residue measurement test for bakers & medics

E. To apply solutions and appropriate health labelling for food manufacturers

CONCLUSIONS