Undenatured Grassfed Whey Protein Powder

What Does Undenatured Mean?

What is “Undenatured” or “Non-Denatured” Protein?

This article aims to clarify a few trending buzz words used to both describe and too often market whey protein. Whey protein is said to be "denatured" the moment its structure deviates from its native conformation.

Denaturing occurs as a function of the proteins' over-exposure to specific elements capable of altering any of the four protein structures. Factors capable of denaturing proteins include but are not limited to:

• Hot & Cold Temperatures
• pH Extremes
• Ultra-Violet Exposure (Sunlight Radiation)
• Mechanical Agitation

In searching storefronts in hopes of finding the best grass-fed whey protein, you will most likely spot a label that states something to the effect, "Undenatured Whey Protein" or even "Non-denatured Whey Protein." This claim is becoming as popular as "Cold Processed" & "Grass-Fed."

The idea behind the marketing point is a suggestion that careful sourcing, manufacturing, and handling is given to the protein to ensure the protein structures are entirely intact. This single adjective markets the product’s ability to transfer its nutrients to you. This claim also leads to an inference that the other competing brands who are not claiming this front & center may be providing a whey protein that is less bio-available or not bio-available at all.

Learn More: What is "Cold Processed" or "Cold Pressed?"
Learn More: Does Cooking with Whey Protein Denature it?

What Does Undenatured mean?

As we examine the various points in the whey protein manufacturing process where whey protein has the potential to be rendered nutritionally ineffective, we’ll use several descriptive words & phrases.

Phrases like, “damaged protein,” “denatured peptides,” and “altered whey protein fractions” for the sake of this article, all refer to the same concept; whey protein and specific protein fractions are fragile and susceptible to being deformed.

When the second, third, or fourth protein structures become permanently deformed, the body may have difficulty recognizing its nutritional value. The proteins in the digestive system known as enzymes, and their ability to navigate these situations depend a lot on your gut health paired with the severity of denaturation of the protein consumed.

If the 1st structure, known as the primary protein structure is permanently damaged, the individual amino acids may easily be deformed as well, thus drastically reducing the chances of the protein being utilized by the body in any capacity.  

The degree or severity of the “denaturing” depends solely on the specific denaturing element, with the magnitude of the force and its exposure time being the critical factors to the whey protein’s structural integrity. 

Note: It’s essential to understand that undenatured whey protein powder can quickly become denatured or further denatured long after it’s packaged and shipped via Amazon Prime to your residents.

Denaturing whey protein isolate isn’t exclusive to the manufacturing process but rather can occur at any time. “Non-denatured whey protein” is another term that means “Undenatured whey protein.”

Learn More: What Are Protein Fractions 

• Bovine Serum Albumin (BSA)
• Immunoglobulins (Ig)
• Alpha Lactalbumin (α-La)
• Beta Lactoglobulin (β-Lg)
• Lactoferrin
• Glutathione

What does Denatured Mean?

"Denaturing" when referring to proteins breaking down and reorganizing a protein's structure is what we call protein denaturation and can occur through various methods. As mentioned, the heat, acid, and mechanical action of your digestive system all denature proteins.

Denaturation is sort of a natural part of life for a protein and is somewhat unavoidable. But, does it make your protein worthless from the point of view of bioavailability?

The short answer is, it depends entirely on the severity of the structure change and or damage. If the slightest denaturation of protein did cause bioavailability issues, humanity would be in a pickle.

The scientific community has yet to pass judgment based on immutable evidence on protein denaturation impacts and the sensitivities that matter most; however, the consensus with critical mass believes less is ideal. To explore this further, let's discuss "Bioavailability".

What Is Bioavailability?

Bioavailability is a buzzword like many others in the nutrition and dietary supplement industry that lacks a clear and well-defined scientific definition. The term originates in the pharmacology community, defined as the rate at which a drug can reach the targeted action site.

For protein powders, however, the pharmacology definition of bioavailability falls short of meaningful. The pharmacology term is ineffective when applying it outside the industry due to the measurement not considering the function of the user's nutritional status and physiological state.

In other words, bioavailability for nutrition is very specific to the individual processing of the particular macro (carbohydrate, protein, fat). There are incredible custom dieting services available that can tailor a diet and then evolve it taking into account your body's physiological response. Your body's response is a reflection of the bioavailability of the nutrients you are taking on.

Applying "Bioavailability" to Whey Protein Powder

A definition most widely used in discussions around protein powder is that bioavailability is a measure of the macro's capacity of being absorbed for the body's immediate use or storage, as a proportion of its mass consumed.

The Connection of Gut Health and Bioavailability

The health of the consumer's digestive tract plays a vital role in determining a protein powder's bioavailability. When the gut is healthy, various enzymes thrive.

These enzymes are responsible for deconstructing the consumed protein and sorting the amino acids such that your body can absorb them. Below are two gut health scenarios to consider to help drive home why gut health is crucial in the bioavailability conversation.

1.) Your gut is functioning optimally -

• High-quality undenatured protein - Bioavailable
• Low-quality denatured protein - Partially Bioavailable

• High-quality undenatured protein - Partially Bioavailable
• Low-quality denatured protein - Not Bioavailable

In Summary, the body's protein cleavers (enzymes responsible for sorting amino acids) will either allow the body to piece together a highly deformed protein making it work, or reject it as unrecognizable.

By focusing on things easily controlled and choosing a less damaged protein if given the option, will increase your body's chances of absorption and, thus, bioavailability.

Learn More: Whey Isolate Digestive Enzymes & Bioavailability
Learn More: Protein Bioavailability - Explained 

What Causes Denaturation of Whey Protein? 

The term “denatured protein” refers to a protein compound (made up of several amino acids connected by peptide bonds) whose spatial arrangement is no longer natural.

This alteration process that physically changes the shape of the protein compound is known as “denaturation.” Many forces have the potential to denature your whey protein. Some of these forces exist during the manufacturing process, while others, incidentally applied after the protein is in the consumers’ possession. 

The following elements comprise the primary forces that cause protein structures to change form (denature):

• High Heat / Temperature (161°F is the limit to be safe) 
• Pasteurization Method (Non-denatured whey protein must be pasteurized via the Vat or HTST method only)
• Mechanical Shear (Aggressive shaking or blending)
• Chemicals (Acids used in “Ion Exchange Method” of making WPI) 
• Ultra Violate rays (Sunlight)

What Part of the Protein Powder Becomes Denatured?

High Temperatures or Ultra Pasteurization damage the tertiary and quaternary protein structures -

Mechanically Shearing (Blending) your protein shake to a frothy foam air smoothie almost guarantees tertiary & quaternary denaturing.

Chemicals used in the Ion Exchange Method aren’t as harmful as most “cold-processed” (marketing) focused whey protein brands would have you believe.

Most studies found that of the major whey proteins, α-lactalbumin, and β-lactoglobulin, both mostly go unaffected by the pH adjustment needed for the Ion Exchange method to work [3].

Ultra Violet rays (Sunlight) primarily damages the tertiary structure. 

What Are the Whey Protein Structures that Become Denatured?

At What Temperature Does Whey Protein Become Denatured -

Answer: Milk pasteurization standards (IDFA – International Dairy Foods Association) across the globe call for a minimum temperature for Vat & HTST Pasteurization.

• Vat Pasteurization requires heating the milk for 30 min @ 145°F.
• HTST (High-temperature short-time) Pasteurization requires heating the milk for 15 seconds @ 161°F.

We consider both Vat & HTST “cold-processing” pasteurization methods. Each method optimizes the milk for both the inactivation of bacteria (pathogens) and the preservation of the protein structure (protein fractions remain intact). 

Science shows that proteins, in general, begin to denature at 40°C (104°F). When milk undergoes pasteurization, denaturation does indeed occur, albeit reversible denaturation. When the milk cools, and the heating element is no longer a denaturing force, the protein structures reform as if the denaturation never happened.

Studies over the years have shown that β-lactoglobulin (LG) is one of the most sensitive fractions to temperature-related denaturing. The research concludes that either of the “cold-processing” pasteurization methods will leave the protein structures unaltered due to temperature [1]. The study referenced looks at both β-lactoglobulin (LG) & α-lactalbumin (LA) -

• Temperatures < 70°C (158°F) - Results in no Denaturation of (LG) & (LA)
• 70°C (158°F) < Temperature < 80°C (176°F) - Denaturation of (LG) & (LA) begin, however the severity is time-dependent.
• Temperatures > 80°C (176°F) - Results in Denaturation of (LG) & (LA) after only 15 seconds

What Pasteurization Methods Assure an Undenatured or Non-Denatured Whey?

Although the cold-processing pasteurization methods (Vat & HTST) will ensure your whey protein powder is flush with nutrients able to be absorbed, these are generally less common practices. Across the industry, the current methods trend towards the HHST (Higher-Heat Shorter Time) methods, which allow for even shorter exposure times down to .01 seconds but at temperatures that immediately disrupt the whey protein structures rendering the end product, stable but “denatured” to say the least.

The overwhelming majority of whey protein and supplement brands have limited visibility to supply chain elements upstream of their ingredient company to whom they are in direct contact. These brands have no idea where their whey protein powder originates, let alone if it’s from grassfed farms. It’s a good practice to have that correspondence with any brand making such detailed claims. These claims include Grassfed, Native, Raw, Naked, Undenatured, etc.

To Learn More about Native Whey Protein - What is Native Whey Protein?

What Pasteurization Methods Denature Whey Protein -

As detailed above, if the pasteurization method exposes the milk to sustained temperatures above 161°F, the protein fractions will begin to denature. The ways that coincide with higher temperatures than 161°F fall into the HHST (Higher Temp Short Time) and the UP (Ultra-Pasteurization) category.

These HHST methods expose these fragile proteins to temperatures as harsh as 100°C (212°F). As you can see from the data above, it doesn’t take much heat to begin the denaturation process.  

If you are relying on whey protein powder for specific macro targeting, you may not be hitting the numbers needed to satisfy your body’s needs. Study after study on HHST pasteurization methods, suggests structural modifications (denaturing) occur in the proteins, especially those belonging to the casein micelles.

For whey protein, the attraction for many consumers resides in the branched-chain amino acid concentrations. Where a consumer may value nutrient density and bioavailability, a manufacturer may value shelf life and stability. Unlike cold-processing methodologies that are optimized to balance microbial lethality points (safety) and nutrition bioavailability (non-denaturing). The alternative methods of pasteurization available to maximize shelf life and product stability, sacrifice nutrient content. The alternative techniques fall in the HHST category but also include the UP (Ultra High-Temperature Pasteurization) process.

The UP method exposes the raw milk to temperatures upward of 280°F. This process allows continuous flow during the pasteurization process and optimizes the output of a factory required to run around the clock - 24/7. These drivers are endemic to countries like New Zealand, whose dairy production serves global demand (Mostly China) as the largest exporter in the world. The faster companies like Fonterra routinely process, then ship dairy across the ocean; the longer the product stays stable (shelf life), the better for all involved, except the consumer.

“Raw” Whey Protein is a Myth -

Although the “degree” of structural alterations can matter if you are using a high-end grassfed whey protein product loaded with macro-nutrients, consuming “raw” milk can be wildly dangerous with severe consequences.  Subsequently, consuming whey protein derived from unpasteurized milk can be deadly. 

Inducing Mechanical Denaturalization -

Whey proteins, in general, are amphiphilic. They contain some elements that like water and other features that prefer oil.  Many brands add an emulsifier to reduce the surface tension of the protein in solution. This practice allows for easy (no clumps) stirring or mixing with water or other choice liquids.  Another way to reduce surface tension is to shear it with a blender or similar mechanical means.

Whey Protein with sunflower oil (Sunflower Lecithin) like AGN Roots Grass-fed Whey Isolate due to its hydrophilic nature (ability to bind efficiently to water) is said to be “instantized” (ready-to-mix).  It will perform well in terms of mixing (solubility) into solutions over a full window of pH values, which is why whey protein recipe applications can cover everything from ice cream to pizza.

Note: Whey protein that is instantized using an emulsifier such as lecithin is more susceptible to mechanical denaturation due to the reduced surface tension. Single-ingredient whey protein, also vulnerable to being denatured mechanically, does require more agitation than instantized whey due to its natural proclivity to resist dispersing in water.

When blending a whey protein solution (shearing with blades), the speed of the blender is critical to maintaining an undenatured whey protein powder.  Reducing the blender speed as much as possible to achieve dispersion is ideal.

If the solution begins to foam, this is a sign that air has forced its way into the protein structure. Air molecules are not polarized one way or the other, which means they sit adjacent to the protein structure and apply an unnatural force like a puzzle piece that doesn’t fit.  Eventually, if the energy is too high, the puzzle pieces break, this analogy represents mechanical denaturation. 

Specifically, it’s the tertiary and quaternary structure of the whey protein molecule that gives way with the forced insertion of air molecules.

What is Protein Renaturation?

Renaturation occurs when a denatured protein reverts to its original structure after the denaturing force is no longer relevant. An excellent example of renaturation not being possible is applying heat to an egg white. Once the egg goes through irreversible denaturation, there is no going back.

On the other hand, if we think about mechanical denaturation and a whey protein shake foaming like champagne after undergoing a vigorous shaking. If the protein sits long enough and the foam returns to a homogenous solution, chances are that a fair amount of protein renaturation is occurring.

Also, applying heat to milk above 40°C (104°F), as discussed earlier, causes denaturation. Keeping these temperatures below 161°F, however, allows this denaturation to be reversible. When the heating element no longer poses a threat, the milk cools the structures' reform.

The Primary-Structure is like the DNA of proteins; within, the blueprints of the secondary, tertiary, and quaternary structures reside. As long as the Primary-Structure isn't damaged, the protein's bioavailability potential remains.

How to Avoid Mechanically Denaturing your Undenatured / Non-denatured Whey Protein Isolate -

If your protein is already instantized (contains lecithin), blend all smoothie ingredients first, then stir a protein solution on the side (protein powder + water or milk of choice already mixed with a spoon). Finally, mix the blended smoothie ingredients and the protein solution as the final step.

If accustomed to using a single-serve blender with an air-tight seal like a Magic Bullet or Ninja mixer, limiting the amount of air in the container will cut the air supply down to negligible levels and allow you to blend away.

In all cases, the idea is to reduce the amount of exposure the whey protein has to mechanical shearing.  Both the speed of the shearing force (energy) and the time under this tension are critical to ensure your undenatured whey protein stays that way.

Ultra Violet (UV) Rays Will Denature Whey Protein

For the same reason, beer and wine are most often packaged in dark-colored glass (green & browns) with either a UV barrier or other form of UV protection within the glass itself; whey protein needs protection from this destructive element too.  The severity of this denaturation due to UV ray exposure, in general, is less than the associated temperature that may come with this exposure.  Where there is sunlight, there is likely heat. 

Naturally, you wouldn’t have an attraction to a beautiful wine bottle that is clear plastic or transparent, see-through glass.  The same logic applies to whey protein. Brands that house fragile whey protein in giant clear plastic tubs and market with the terms “Undenatured or Non denatured Whey Protein” should make you wonder what else amongst their processing is lacking understanding and care.

Studies have shown that the Tertiary Structure of protein is susceptible to denaturation due to sunlight. The damage UV rays impart impact the α-lactalbumin fraction more severely than the β-lactoglobulin fraction; however, damage to both is likely, and it will render their nutritional bio-availability ineffective [2].

~AGN Roots Grassfed Whey Team

References

[1] β-Lactoglobulin is a Thermal Marker in Processed Milk as Studied by Electrophoresis and Circular Dichroic Spectra
Chen, W.L. et al.
Journal of Dairy Science, Volume 88, Issue 5, 1618 - 1630

[2] Effects of ultraviolet radiation on properties of films from whey protein concentrate treated before or after film formation
Olga Díaz, Davinia Candia, Ángel Cobos
Food Hydrocolloids, April 2016

[3] Effect of pH on the Association of Denatured Whey Proteins with Casein Micelles in Heated Reconstituted Skim Milk
J. Agric. Food Chem.2003,51, 6, 1640-1646
Publication Date: February 13, 2003