The Protein BridgeLinking Protein Structure to Function and Application

Julie Emsing MannMay 6, 2015

Protein and Hershey

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Overview

• Protein Selection• Structure Determines Functionality• Functionality Determines Application• Application Examples• Overview of Current and Emerging Protein Sources

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Protein Selection

• Protein concentration• Protein functionality desired• Protein fortification desired• Protein quality (PDCAAS)• Protein sourcing/availability• Clean label production• GMO status• Sustainability • Consumer Acceptability• Allergenicity• Kosher requirements• Price/lb

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Protein Selection

Protein Primary Functionalities Limiting Attributes

Dairy Emulsification, Gelation Cost volatility, Lactose

Soy Emulsification, Structure Soy flavor

Gelatin/Collagen Gelation, Water-binding Pork/beef derivation

Peanut Flavor, Texture Aflatoxin concerns

Almond Flavor, Texture Cost

Wheat Film Formation, Structure Gluten presence

Egg Aeration Unstable to heat

Muscle Binder, Gelation Meat processing/Safety

Bean Binder, Viscosity Bean flavor

Algal Gelation, Emulsification Consumer Perception Unknown

Canola Binder Consumer Perception Unknown

Pea Emulsification, Binder Pea flavor

Rice Bran Film Formation, Binder Consumer Perception Unknown

Traditional

Emerging

Protein Structure

Protein Types Determine Function in Food and Beverage

Protein Structure Dictates Function in Food and Beverage

Amino acid Basic unit

•Differ by their side chains •Charged

PeptideShort chain of AA’s

•Joined by peptide bond

ProteinPolymers that fold into a 3D structure

http://www.chemicalconnection.org.uk/chemistry/topics/view.php?topic=5&headingno=3

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Amino Acid Map

Food Chemistry, 20059

Peptide bonds

Bond type

H-Bonds

Hydrophobic,Di-sulfideIonic

Hydrophobic

http://www.ucl.ac.uk/~sjjgsca/ProteinStructure.html

Protein Structure Influenced by Treatment/Environment

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Protein Denaturation Affects Structure

DENATURED STATE

• Loss of native conformation• Altered secondary, tertiary or quaternary

structure• May be reversible or irreversible, partial

or complete

• Results• Decrease solubility• Increase viscosity• Altered functional properties• Loss of enzymatic activity

NATIVE STATE

• Usually most stable and soluble• Polar groups usually on the

outside• Hydrophobic groups on inside

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Denaturing Agents

Temperaure pH Shear

High Pressure Processing

Organic Solvents Salts

Oxidizing and Reducing

Agents

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Beyond the Specification Sheet

• Basic spec sheet information is not sufficient to understand protein ingredients.

• Ask suppliers if they can provide you information on:• How the protein was processed

• Heat• Filtration

• Degree of denaturation through processing

Protein Structure Functionality

Knowing the structure and degree of denaturation in selected protein source is fundamental to predicting its functionality. The environment plays a role as well.

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Protein Structure is Highly Dependent on Environment

pH Temperature

Dielectric constant

Ionic strength

Other Molecules Air

Fat Denaturing Agents

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Proteins Strive to Achieve Lowest State of Free Energy

Hydrophillic

Hydrophobic

Protein in Aqueous Solution (Native)

Isolated Protein

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Protein Functionality in Food/Beverage Systems

Protein Ingredient Functionality in Food

Water Binding Viscosity Gelation

Foaming Emulsification Browning

Flavor/texture Protein fortification/nutrition

Protein Functionality Importance

• Shorten development time• Aid scale-up efficiency• Save $$$• Ensure stable products through-

out shelf life• Provide unique function leading

to novel products

Understanding protein

selection and functionality for specific

projects can:

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Interactions Effecting Food Protein Function

1) Protein/Water interactions• Hydration of protein

• Solubility, water absorption, wettability, viscosity

2) Protein/Protein interactions• Within or between proteins

• Network building, precipitation, gelation

3) Protein/Other interactions• Air, fat, carbohydrates

• Surface tension, emulsification, foaming/aeration, film forming

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Protein Solubility

• Functional properties of proteins depend on their solubility

• Charged amino acids have primary role in protein solubility• The proteins are least soluble at their

isoelectric point

• The proteins become increasingly soluble as pH is increased or decreased

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Critical Parameters to Understand Functionality

The Protein1. Protein type/structure

Solubility: Hydrophillic and hydrophobic exposure

2. Protein concentrationMore protein = more water uptake, molecules closer together

3. Protein denaturationHow much protein is denatured pre-process?

The Food/Bev System1. pH

• Acidic, neutral or basic

2. Interactions with other compounds:

• Proteins• Carbohydrates• Salts/Minerals

The Process

1. Heat treatment

2. Shear

3. Pressure

Protein + Environment + Process =Control of Desired Properties

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Function to Application

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Functionality per Application

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Solubility- Beverage Example

• Primary proteins utilized: • Dairy, Soy

• Production• Hydration of protein and hydrocolloid, other

ingredients• Heat treatment• Homogenization• Filling

• Chemistry• Stabilization of protein with hydrocolloid

interaction• Homogenization reduces particle size

Beverage Sample Formula

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Gelation- Gummy Bear Example

• Primary Proteins Utilized: • Gelatin

• Production:• Boiling of sugar/corn syrups, addition of

gelatin, removal of water, cooling and forming/depositing in starch molds to remove surface moisture.

• Chemistry:• Collagen molecules are mobile in hot

water, as cooling occurs, they are restricted

• Junction zones form and bond the molecules together, forming a gel network

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Gummy Bear Sample Formula

Ingredients %

Gelatin-Type A 9.00

Water 15.00

Sucrose 32.00

Corn Syrup (43 DE) 42.00

Citric Acid Solution 1.00

Flavor 0.50

Color 0.50

Total 100.00

Procedure: (1) Hydrate gelatin in hot water, cool to form gelatin plug, cut into pieces(2) Combine and boil sucrose, corn syrup to reach X solids (temp)(3) Add pieces of gelatin to cooked syrup, mix well(4) Add remaining flavor, color, acid(5) Deposit into starch molds

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Foaming-Marshmallow Example

• Primary Proteins Utilized: • Egg albumen, gelatin, hydrolyzed soy

or milk• Production

• Frappe produced: aerated/whipped• Syrup produced, cooked, added to

frappe• Gentle mixing, slab or deposit

• Chemistry • Hydrophobic amino acids orient

towards gas phase• Viscous protein solutions create more

stable final foams

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Marshmallow Sample Formula

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Protein SourcesThe Old and The New

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Protein Evolution

Dairy Soy Egg

Wheat

Pea

Canola

Potato

Pulses

AlgaeLemna

RuBisCoInsectsWolffia

Knowledge Building

Dairy

Pros:• Whole food perception• Mixture of casein and

whey• Complete protein• 40-90% protein content• Excellent functionality• Long history of usage

Cons:• Cost volatility• Animal derivation• Consumer concerns

over antibiotics and hormones in milk

Soy

Pros:• Plant derived• Complete protein• Predominately glycinin and β- conglycinin

• 40-90% protein content• Excellent functionality• Heart health claim• Long history of usage

Cons:• Anti-nutritionals• Off-notes/beany• Perception of

hormone/estrogenic

Gelatin

Pros:• Pure protein• Unique gelling

properties• Heat Stable• Intact or hydrolyzed• Health Linkages:

• Joint health linkage• Beauty from within

• Gluten free

Cons:• Animal source• Not Kosher• Not a complete

protein

Pulses

Cons:• Newer science and

technology• Not complete proteins

Pros:• Plant derived whole

food• Contains fiber and

protein• Sustainable• Many regions of world

already rely on this in their diet

Wheat Protein

Cons:• Not a complete protein

• Low lysine

• Gluten sensitivity

Pros:• Unique functionality

from gliaden and glutenin

• Elastic properties can not be duplicated with other proteins

• Bland flavor profile• Low cost

Emerging Protein SourcesSource Forms Available Maximum

ProteinSupply Sufficiency

Algae Flours: high protein or high lipid

55% Medium

Canola Flour, isolate, hydrolysate 90% Small/Medium

Oats Flour, concentrate 55% Small/Medium

Flax Meal, concentrate, crisps 50% Med/Lg

Hemp Seed, flours, concentrates 50% Small

Quinoa Seed, crisps, flours 65% Small

Rice Flour, concentrates, crips 80% Medium

Sunflower Flour, concentrate 70% Small

Lemna Flour, concentrate 65% Small

• Contain highly functional and nutritional protein.• Consumption areas are predominantly outside of the US.

Live Crickets Dried Crickets Live Meal WormsDried Meal

WormsMoisture, % 69 8 61 6Fat, % 5 15 13 22Protein, % 20 60 20 60Fiber, % 3 8 2 10Ash 2 8 2 1Ca, ppm 345 No data available 133 283P, ppm 4238 No data available 3345 2161

Insect protein

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RuBisCo

The most abundant protein found on earth and presentin every green, leafy material.

What is it?• Ribulose-1,5-bisphosphate carboxylase/oxygenase• Enzyme in the first step of carbon fixation for photosynthesis

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