What does "Undenatured" or "Non-Denatured" Protein mean?

This article aims to explain some commonly used terms related to whey protein, specifically "undenatured" or "non-denatured" protein. When whey protein is denatured, it means its natural structure has been altered.

Denaturation can occur when proteins are exposed to certain elements such as high or low temperatures, extreme pH levels, sunlight radiation, or mechanical agitation.

If you're looking for the best grass-fed whey protein, you might come across labels claiming "Undenatured Whey Protein" or "Non-denatured Whey Protein." These terms have become popular marketing points, similar to "Cold Processed" and "Grass-Fed."

The idea behind this marketing claim is that the protein has been carefully sourced, manufactured, and handled to ensure its structure remains intact. The use of this adjective suggests that the nutrients in the protein can be effectively transferred to you. It also implies that other brands not making this claim may offer a whey protein that is less bioavailable or not bioavailable at all.

To understand what "undenatured" means, let's explore the various stages of whey protein manufacturing where it can lose its nutritional effectiveness. Terms like "damaged protein," "denatured peptides," and "altered whey protein fractions" all refer to the same concept: whey protein and its specific fractions are delicate and prone to deformation.

When the second, third, or fourth protein structures are permanently deformed, your body may struggle to recognize the nutritional value of the protein. The enzymes in your digestive system that handle proteins rely on your gut health and the extent of protein denaturation.

If the primary protein structure is permanently damaged, the individual amino acids may also become deformed, greatly reducing the chances of the protein being utilized by your body.

The extent of denaturation depends on the specific element causing it, with the force applied and the duration of exposure being crucial factors for the whey protein's structural integrity.

It's important to note that undenatured whey protein powder can become denatured or further denatured even after it's packaged and shipped to your doorstep.

Denaturation of whey protein isn't exclusive to the manufacturing process but can happen at any time. "Non-denatured whey protein" is another term that means the same as "undenatured whey protein."

If you want to learn more about protein fractions, here are some examples:

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

 

What Does Denatured Mean?

When we refer to protein denaturation, we mean the process of proteins breaking down and reorganizing their structure. Denaturation can happen through various methods, including heat, acid, mechanical action and during digestion. It is a natural occurrence for proteins, but the extent of denaturation and its impact on bioavailability vary.

The severity of denaturation determines whether it affects the protein's bioavailability. If even a slight denaturation negatively impacted bioavailability, it would be a significant concern. However, the scientific community has not reached a definitive conclusion on the impact of protein denaturation on bioavailability. While many experts believe that less denaturation is ideal, the sensitivity to denaturation varies among individuals.

 

What Is Bioavailability?

Bioavailability refers to the body's ability to absorb and utilize a particular nutrient, in this case, protein. In the context of protein powders, bioavailability measures how well the protein can be absorbed and used by the body compared to the amount consumed. It is important to consider an individual's nutritional status and physiological state when assessing bioavailability.

The health of your digestive system plays a crucial role in determining how much protein from a protein powder can be effectively absorbed by your body. When your gut is healthy, it contains various enzymes that are responsible for breaking down the protein you consume and organizing the amino acids in a way that allows your body to absorb them.

 

The Importance of Gut Health and Bioavailability

To illustrate the significance of gut health in relation to bioavailability, consider the following scenarios:

Optimal gut function:

• High-quality undenatured protein: Highly bioavailable
• Low-quality denatured protein: Partially bioavailable

Sub-optimal gut function:

• High-quality undenatured protein: Partially bioavailable
• Low-quality denatured protein: Not bioavailable

The enzymes in your body that break down protein can either attempt to piece together a heavily damaged protein and make it usable, or they may reject it because it is unrecognizable and cannot be processed effectively.

By focusing on factors that you can control, such as selecting a less damaged protein when given the option, you can increase the likelihood of your body absorbing the protein and, consequently, improve its bioavailability. Put simply, quality matters.

 

What Causes Denaturation of Whey Protein?

Denaturation refers to the process of altering the natural shape of a protein compound, which is made up of amino acids connected by peptide bonds.

There are various factors that can denature whey protein. Some of these factors occur during the manufacturing process, while others may occur unintentionally after the protein is purchased by consumers.

The primary forces that cause denaturation of protein structures are:

• High Heat/Temperature: When whey protein is exposed to high temperatures (above 161°F), its tertiary and quaternary structures are affected. The natural coils and folds of the tertiary structure begin to change and unfold as the temperature increases. Vibrations within the molecular bonds holding the quaternary structure together become more intense. Ultimately, these bonds fail due to the stress from the vibrations, leading to protein denaturation. Pasteurization, which involves applying heat for a specific duration, can cause similar damage to the tertiary and quaternary structures.
• Mechanical Shear: Aggressive shaking or blending, such as when making a frothy protein shake, can cause denaturation of the tertiary and quaternary structures.
• Chemicals: Some whey protein brands use acids in the "Ion Exchange Method" of production. However, the chemicals used in this method are not as harmful as certain brands may suggest. Research shows that the pH adjustment required for the Ion Exchange method does not significantly affect the major whey proteins, such as α-lactalbumin and β-lactoglobulin.
• Ultraviolet (UV) Rays (Sunlight): Exposure to UV rays primarily damages the tertiary structure of whey protein.

Factors like high heat, mechanical shear, chemicals, and UV rays can cause denaturation of whey protein. High temperatures affect the tertiary and quaternary structures, mechanical shear disrupts the tertiary and quaternary structures, chemicals used in certain methods have minimal impact on the protein, and UV rays primarily affect the tertiary structure.

 

What happens to whey protein structures when they become denatured?

When milk is pasteurized, it is heated to specific temperatures to kill bacteria and preserve its protein structure. There are two common pasteurization methods: VAT Pasteurization, which involves heating the milk for 30 minutes at 145°F, and HTST (High-temperature short-time) Pasteurization, which requires heating the milk for 15 seconds at 161°F. Both methods are considered "cold-processing" because they aim to inactivate bacteria while keeping the protein intact.

Proteins generally start to denature at around 40°C (104°F). During pasteurization, denaturation does occur, but it is reversible. Once the milk cools down and is no longer subjected to heat, the protein structures reform as if denaturation never happened.

Studies have found that one of the most sensitive protein fractions to temperature-related denaturing is β-lactoglobulin (LG). Research shows that both Vat Pasteurization and HTST Pasteurization, with their specific temperature ranges, do not alter the protein structures due to temperature. Here are the findings:

• Temperatures below 70°C (158°F) do not result in denaturation of β-lactoglobulin (LG) and α-lactalbumin (LA).
• When the temperature is between 70°C (158°F) and 80°C (176°F), denaturation of β-lactoglobulin (LG) and α-lactalbumin (LA) begins, but the severity depends on the duration of exposure.
• Temperatures above 80°C (176°F) cause denaturation of β-lactoglobulin (LG) and α-lactalbumin (LA) after only 15 seconds.

 

Which pasteurization methods ensure that whey protein remains undenatured? 

Cold-processing pasteurization methods, such as Vat and HTST, are less commonly used but can maintain the nutrient content of whey protein powder, allowing for easy absorption. However, the industry currently leans towards HHST (Higher-Heat Shorter Time) methods, which involve very short exposure times (as low as 0.01 seconds) at higher temperatures. Although this method stabilizes the end product, it disrupts the structure of whey protein, resulting in a denatured product.

Unlike brands such as Go Good who maintain control over all aspects of the production process most whey protein and supplement brands have limited knowledge about the origins of their whey protein powder - typically selling ‘white labeled’ supplements.

As a result they are often unaware of whether it comes from grass-fed farms or have visibility into their supply chain beyond their direct contact with an ingredient company who manufactures on their behalf.

 

How Pasteurization Methods Affect Whey Protein Denaturation

If milk is exposed to sustained temperatures above 161°F during the pasteurization process, the protein fractions, specifically whey protein, will start to undergo denaturation. There are two pasteurization methods that involve higher temperatures than 161°F: HHST (Higher Temp Short Time) and UP (Ultra-Pasteurization).

HHST methods subject the fragile proteins to temperatures as high as 100°C (212°F). As shown in the data above, even a small increase in temperature can initiate the denaturation process.

If you rely on whey protein powder to meet your specific nutritional goals, you may not be getting the desired protein content your body needs. Numerous studies on HHST pasteurization methods indicate that structural changes, or denaturation, occur in the proteins, particularly those found in casein micelles.

Many consumers are drawn to whey protein due to its high concentrations of branched-chain amino acids. While consumers value nutrient density and bioavailability, manufacturers prioritize shelf life and product stability. Cold-processing methods are designed to balance safety (eliminating harmful microbes) and nutrition bioavailability (preserving protein structure). However, alternative pasteurization techniques, including HHST and UP, are employed to extend shelf life and product stability, but at the expense of nutrient content.

 

“Raw” Whey Protein is a Myth

The belief in consuming "raw" whey protein derived from unpasteurized milk is a misconception. While the degree of structural alterations may matter for high-quality grass-fed whey protein products that contain essential macronutrients, consuming raw milk can be extremely dangerous, with severe consequences. Therefore, consuming whey protein obtained from unpasteurized milk can be life-threatening.

 

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 Go Good 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?

Protein renaturation happens when a protein that has been unfolded or altered in structure goes back to its original shape once the factors causing the alteration are removed. A simple example to understand when renaturation is not possible is heating egg white. Once the proteins in the egg white are denatured irreversibly, they cannot return to their original structure.

On the other hand, let's consider mechanical denaturation, like vigorously shaking a whey protein shake until it foams like champagne. If the protein shake sits for some time and the foam settles back into a smooth mixture, it is likely that a significant amount of protein renaturation has occurred.

In a similar way, heating milk above 40°C (104°F) causes denaturation, as we discussed earlier. However, if we keep the temperature below 161°F, this denaturation can be reversed. Once the heat source is removed, the milk cools down and the protein structures reform.

The primary structure of a protein is like its DNA. It contains the blueprints for the secondary, tertiary, and quaternary structures. As long as the primary structure remains undamaged, the protein retains its potential for bioavailability.

 

How to Keep Your Whey Protein in its Natural State:

If your protein powder is already instantized and contains lecithin, follow these steps to avoid damaging its quality. First, blend all the ingredients for your smoothie together. Then, separately mix the protein powder with water or your preferred type of milk using a spoon. Finally, combine the blended smoothie ingredients with the protein solution.

If you typically use a single-serve blender with an airtight seal, like a Magic Bullet or Ninja mixer, minimizing the amount of air in the container will greatly reduce the air exposure during blending.

In all cases, the goal is to minimize the whey protein's exposure to mechanical shearing. Both the speed and duration of the blending process can affect the protein's integrity and quality.

 

The Negative Effects of UV Rays on Whey Protein:

Similar to beer and wine, which are often packaged in dark-colored glass containers (such as green or brown) with UV protection, whey protein also requires protection from UV rays. Exposure to UV rays can cause denaturation, although the impact is generally less severe than that of high temperatures, which often accompany sunlight.

Just as you wouldn't be attracted to a wine bottle made of clear plastic or transparent glass, whey protein should also be protected from light. Brands that package delicate whey protein in large clear plastic tubs and advertise it as "undenatured" or "non-denatured" should raise concerns about their understanding and care in the processing of the product.

Research indicates that UV rays can cause denaturation of the tertiary structure of proteins. The α-lactalbumin fraction is more susceptible to damage than the β-lactoglobulin fraction. However, both fractions are likely to be affected, leading to a loss of their nutritional benefits