- glucan and vital - glucan and vital gluten – isolation, gluten – isolation, properties and properties and applicationsapplications

Jyoti Jha

(L-2006 -A-9-D)

What is beta What is beta glucan?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 Structure unit of - -

glucanglucan• Structural unit

• 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)

Beta Glucan Research Organization

Sources- Betaglocon.org

SourcesSources - glucan- 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-Are barley beta-glucans better?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 Isolation and purification of beta purification of beta

glucansglucans Flour(1g)

40 ml MES/TRIS buffer (pH 8.2)

Sequential hydrolysis with

-amylase

protease

Amyto-glucosidase

Hydrolysis

Centrifugation10,000 RPMFor 15 mint

Filtering the supernatant (~ 50 ml)

Cont….

Protocol first For isolation

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)

Freeze-drying

Ref- Goosong et al 2000

Hurray..

How do beta-How do beta-glucans work ?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

-Bile acids

-Glucose

ResultReduces glucose in blood

Reduces insulin level

Cholesterol-lowering

Cont…

(in the small intestine )

In the large intestine- beta-glucan is a prebiotic

stimulate

growth of desirable bacteria limits the growth of potentially harmful organisms

bacteria ferment beta-glucan to produce volatile fatty acids

protect against bowel disease.

Properties ofProperties of - - glucanglucan -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 Application of - - glucanglucan 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…

Function in the Body.

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.

In immune system

Absorption

Dietary sources

Now magic of gluten

What is 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

• 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.

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 ...

www.ndmill.com

GluteGlutenn

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 How to separate the gluten from wheat flour ?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 ProteinsWheat Flour Proteins

Dough

Gluten

Glutenins Gliadins

Wash out starch granules

Mix with alcohol/water

SolubleInsoluble

Physical Physical properties of properties of glutenglutenGluten complex

Gliadin (a prolamin)

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 GeneticsGlutenin 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 GeneticsGlutenin 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 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: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