- glucan and vital

gluten – isolation,
properties and
applications
Jyoti Jha
(L-2006 -A-9-D)
 What is beta glucan?
β-Glucans (or beta-glucans) are natural
gum polysaccharides occurring in the bran
of cereal grains, most abundantly in barley
and oats and to a much lesser degree in rye
and wheat. ...
 Structure unit of -

• Structural unit
glucan

• Molecular configuration:
1-Polysaccharide
2-Chain of glucose molecules.
3-Glucosidic bond among monomer as -1-4 and -1-3 fashion.
4-The “native” beta-glucans tend to have a higher MW (up to 5400 kDa)

Sources-
Beta Betaglocon.org
Glucan Research
Organization
Sources - glucan
Sources percentage
Yeast > 10
Mushrooms 10-12

Barley 7-10
Oats 3-7
Rye 2-3
Wheat <1

Ref. Betaglucan.com
Are barley beta-glucans
better?
1-Barley is the richest cereal source of beta-
glucans.
2- grain kernels containing about 3-11% by
weight, than oat(3-7%) and wheat(<1%).
3-beta-glucans from high-producing strains
of barley have been shown to be more
effective at lowering blood glucose than
oat beta-glucans.
 Isolation and purification
of beta
Protocol first glucans
Flour(1g)
For isolation
40 ml MES/TRIS buffer (pH 8.2)
-amylase
Sequential hydrolysis with protease

Amyto-glucosidase

Hydrolysis

10,000 RPM
Centrifugation
For 15 mint

Filtering the supernatant (~ 50 ml)

Cont….
Concentration of supernatant using a rotary evaporator (~10 ml)

Passing the concentrated sample through a Sep-Pak C18

Passing through a bio-gel P-6 column (1.5-80 cm)

(mobile phase-degassed distilled water, flow rate:0.33 ml/min, 7min/tube)

Collecting soluble dietary fiber (Beta glucan) Hurray.

.
Freeze-drying

Ref- Goosong et al 2000

How do beta-glucans
work ?
 Strengthens immune system.
Enhancing the ability of –
- macrophages

- neutrophils

- natural killer cells

 Fight a wide range of challenges such as,

Bacteria viruses, fungi and parasites.

Cont…..
Slows down the absorption of -

-Cholesterol

-Fatty acids Reduces glucose in blood

Result
Reduces insulin level
-Bile acids

-Glucose

Cholesterol-lowering
(in the small intestine )

Cont…
  In the large intestine-
 beta-glucan is a prebiotic

stimulate

limits the growth of potentially

growth of desirable bacteria harmful organisms

bacteria ferment beta-glucan to produce volatile fatty acids

protect against bowel disease.

 Properties of - glucan
 -glucan content increased with temperature but not with pH.
 Viscosity increased with pH, maximum at pH 9.0.
 -glucan increased non-specific host resistance to a variety of bacterial,
viral, fungal and parasitic infections.
 Viscosity and molecular weight of -glucan was reduced at acidic
condition.
 Sucrose increased the viscosity of -glucan at low concentration.
 -glucan reducing the blood pressure and inducing the satiety that can
lead to weight loss.
 Cooking at higher temperature and higher moisture levels influenced the
water solubility of -glucan at room temperature.
 -glucan from brewer’s yeast can be used in food products as
thickening, water holding or binding agent and emulsifying stabilizer

Ref- www.eurasyp.org
Application of - glucan
 Clinical applications.
- Cancer: used as an immuunoadjuvant therapy for
cancer (primary in Japan, 1980)
- Prevention of infection:

- surgery:
-Wound healing: 1-fewer infection
2- reduced mortality
3-stronge tensile strength of scar tissue

Cont…
 In immune system

Absorption
 Function in the Body.
Dietary sources

 Medical applications.
- Beta glucan found in baker’s yeast and certain fungi have
anticancer properties.
-Mushroom derived beta glucan used as adjuncts to chemotherapy.
- beta D-glucan important role in the diagnosis of toxic mycosis caused
by fungi.
- Beta glucan is also promoted as dietary supplement for weight loss.
Now magic of gluten
What is gluten ?
A protein group found in wheat and other
flours that forms the structure of the bread
dough. Gluten holds the carbon dioxide
(CO2) produced by the yeast and expands
during fermentation, and provides the
elasticity and extensibility (stretch) in
bread dough. ...

www.hodgsonmill.com
Vital Wheat Gluten
Vital wheat gluten is the gluten proteins
extracted from wheat flour. This is usually done by
making dough, then washing the starch from
around the ...

• A unique water-insoluble vegetable protein with bulking and elastic

properties.
• Creamy tan colored, free-flowing powder.
• Minimum 75% protein content.
• Primarily used in the baking industry.
• Other uses - meat binder or substitute, stabilizer, thickener and
emulsifier.

www.ndmill.com
 Gluten
Gluten is made up of:
- primarily glutenins and gliadins (~80%)
- starch, lipid and fiber are minor components

- Gluten is formed when water and mechanical energy (as mixing

or sheeting) is added to the flour

- Gluten gives dough its visco-elasticity and provides texture to

end products
Chemical Modification of wheat gluten

I. Gluten becomes soluble in a variety of chemicals: soaps,

detergents, hydrochloric acid, sodium hydroxide, 70% ethanol are
just a few possibilities.

II. Solubilization by deamidation is also used; this can be in acidic or

basic solutions. Removal of the amide group of glutamine results
in a change in the potential ionic charge on the protein.

III. Enzymatic modification is a process that hydrolyzes peptide

bonds and increases the solubility of gluten.
 How to separate the gluten from
wheat flour ?
1. Mix a small amount of flour with just enough water to form a stiff
ball of dough

2. Soak the ball of dough in water for about 30 minutes.

3. Over a fine mesh sieve, and under running water from a faucet,
wash out all the starch.

4. If you bake the ball of wet gluten at about 400 degrees F. until all
the moisture has evaporated and cut the dry gluten in half, you will
see how gluten forms the frame work (structure) of baked products.

Cont..
 Wheat Flour Proteins

Dough

Wash out starch granules

Gluten

Mix with alcohol/water

Insoluble Soluble

Glutenins Gliadins
Physical properties of
gluten
Gliadin (a prolamin)

 Gluten complex
Glutenin (a glutlin)

Gliadins -
-Large group of protein with similar properties.
- Molecular weight of about 40,000.
- Single chain and sticky when hydrated.
- Little or no resistance to extension.
- Responsible for the dough cohesiveness.

Cont…
 Glutenin-

- Heterogeneous group of proteins.

- Very in molecular weight from about 100,000 to million.

- Average of molecular weight is about 3 million.
- Protein is resilient and rubbery but prone to rupture.
- Responsible resistance to extension.
 Glutenin Genetics
Wheat is hexaploid and contains
three related genomes: A, B, & D
each with 7 pairs of
chromosomes numbered 1-7

HMW-GS encoded at Glu-1 loci of

chromosomes 1A, 1B and 1D
- Glu-1A, Glu-1B, Glu-1D

Each locus has multiple alleles

- Alleles not genetically linked
- Any combinations possible
 Glutenin Genetics

- Some combinations of HMW-GS

are associated with stronger
dough and better bread making
quality

- Specific sub-unit effects in

noodles not known
 Key Techniques
- SDS PAGE
- identify high molecular weight glutenin subunits
- Size Exclusion HPLC
- Determine molecular weight distribution
- Protein Content
- Quantity analysis
- Mixograph
- recording dough mixer
- Noodle Textural Analysis
- Physical parameters
Vital wheat gluten can be applied:
 As protein source and binder in pet food &
fish food
 In vegetarian meat replacing products
 For special bread
 In breakfast foods which should be rich in
proteins like gluten bread or biscuits
 In breakfasts cereals
 In snacks like spring roll
Cont….
 In soups & sauces
 In dough products
 As exchangers for milk protein e.g. in
cheese products
 In protein foam products