How much protein are you really getting?

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How much protein are you really getting?

This is the study that the supplement industry doesn't want you to see. With the current proteins tested, here is what your body is actually able to absorb:

  • Milk Protein Absorption: 3.5g/hour
  • Pea Protein Absorption: 3.4g/hour
  • Soy Protein Isolate Absorption: 3.9g/hour
  • Casein Isolate Absorption: 6.1g/hour
  • Whey Isolate Absorption: 8g-10g/hour

What does this mean?

Most athletes look to consume 1.5g of protein per pound of bodyweight per day; this may be more or less dependent on your goals. So, if you weigh 200 pounds, your target is 300g of protein per day. This is usually divided into an average of 6 meals, which means you need 50g of protein per meal. What this study shows is that your body can not absorb that amount of protein in a powdered supplement. Whey Isolate has the best absorption rate of up to 10g an hour at 31% absorption. Therefore, you would need 32.5g of powder to get the 10g of usable protein.

Here is where GENEPRO GEN2 comes in; because of the small molecular size of their protein, it has a 97.8% absorption rate. What this means to you is that a 1 TBSP serving of Genepro is equivalent to 32.5g of regular whey protein. GENEPRO's flavorless protein can be mixed into any beverage (hot or cold), in any food item (even baking and cooking) without changing the taste or consistency. Truly the most convenient protein powderh that can be used by anyone, anytime and anywhere! Link to full study: It is pretty lengthy but the most interesting findings are on page 8.

Genepro Gen2 with Prohydrolase® Clinical Study

Background

Whey protein is one of the richest sources of all of the nine essential amino acids which your body is unable to produce. However, whey protein must first be processed into a useable form for building and recovery of muscle (peptides and amino acids) and it must do so in less than two hours to be absorbed. This is the time from the stomach through the small intestine in which digestion/absorption primarily occurs (Kim SK 522-24). Undigested whey will simply be excreted (wasted amino acids).

The use of SDS gel electrophoresis for protease hydrolysis of proteins has been well documented and chosen as the in vitro method to evaluate whey hydrolysis by proteases. A five amino acid peptide created with the help of New England PeptidesTM to mark bioavailability was mixed with a low molecular weight marker to visualize and determine the size of the peptide fragments created by each protease blend on whey.

The in vitro conditions were designed to mimic the processes that occur in the digestive tract when whey is consumed, including buffering capacity of the drink, pH of the stomach and small intestine, salt conditions, bile salts and transient time. Drinks show a transient time of approximately 30 minutes in the stomach and less than two hours in the small intestine (Kim SK 522-24).

Your body produces proteases of its own including pepsin in the stomach and pancreatic enzymes in the small intestine. Most protein drinks buffer the stomach between 4-6 pH, which greatly reduces the acidity needed to unfold/hydrolyze the proteins and the efficiency of pepsin (Fordtran 645-57;Schaafsma G 1865S-7S). This leaves the bile salts (unfold) and pancreatic enzymes (hydrolyze) as the main means to convert proteins into absorbable peptides and amino acids.

Processing of the whey protein before it is dried can create partially aggregated protein that is more difficult to breakdown by proteases than the native protein. This was confirmed by mixing whey with pancreatin using USP small intestinal conditions resulting in very little break down of the whey (Figures 1 and 2). The pancreatic proteases alone were not enough; further processing of whey by a slight drop in pH and introduction of bile salts resulted in better degradation of whey but still not complete breakdown (Figure 3). However, the Genepro Gen2 w/Prohydrolase protease formulation was able to hydrolyze the whey protein with no additional help and in the time needed for absorption in the small intestine.

Although whey has a high protein digestibility corrected amino acid score (1.0), that score can be misleading. Protein Digestibility Corrected Amino Acid Score (PDCAAS) is a method of evaluating the protein quality based on both the amino acid requirements of humans and their ability to digest it. The formula for calculating the PDCAAS percentage is: (mg of limiting amino acid in 1 g of test protein / mg of same amino acid in 1 g of reference protein) x fecal true digestibility percentage (Schaafsma G 1865S-7S), which means that the protein can be broken down in the large intestine where very little absorption (less than 10%)

occurs. Whey also has a high Biological Value (BV), which is a measure of the proportion of absorbed protein from a food which becomes incorporated into the proteins of the organism’s body. Biological value is determined based on this formula: BV = (Nr / Na) * 100.

Where:

Na = nitrogen absorbed in proteins on the test diet

Nr = nitrogen incorporated into the body on the test diet

However, direct measurement of Nr is essentially impossible. While both PDCAA and BV give you a measurement of the nutritional quality of the protein, they have limits to how much of the protein is actually absorbed and used for protein synthesis. The best way to determine if a whey supplement is being processed so your body can use it is by conducting in vitro and in vivo tests. The following is an example of the testing of whey protein supplement and whether or not it is processed enough to provide benefit to the user.

In Vitro Study:

- Whey hydrolysis visualized by silver stain gel electrophoresis under USP physiological conditions (37°C, pH6.8) of the small intestine for 90 minutes with either 2mg Genepro Gen2 w/Prohydrolase protein or USP pancreatin standard (1%)and a combination of both.

- Whey hydrolysis visualized by silver stain gel electrophoresis densitometry under USP physiological conditions (37°C, pH6.8) of the small intestine for 90 minutes with either 2mg Genepro Gen2 w/ Prohydrolase protein or USP pancreatin standard (1%).

- When whey protein is not broken down into the smallest composition, it creates large peptides that can cause discomfort such as bloating, nausea, cramping, pain, etc. When depending solely on the body’s own pancreatic enzymes, large peptides can be present for long periods of time in the digestive tract. Under the conditions found in the small intestine in the laboratory, Genepro Gen2 w/Prohydrolase breaks down these large peptides (Figure 2) reducing the potential for discomfort that often comes from consuming large amounts of proteins such as whey. This can be shown in vitro using immunoassays that quantify these peptides down to 1ppm (Rosendal A 2200-10). Whey, hydrolyzed by either Genepro Gen2 w/Prohydrolase or Pancreatin and an untreated sports digestive aid alone were examined using this method (Figure 4). The higher the bar, the more “bad” peptides are present.

When whey protein is not broken down into the smallest composition, it creates large peptides that can cause discomfort such as bloating, nausea, cramping, pain, etc. When depending solely on the body’s own pancreatic enzymes, large peptides can be present for long periods of time in the digestive tract. Under the conditions found in the small intestine in the laboratory, Genepro Gen2 w/Prohydrolase breaks down these large peptides (Figure 2) reducing the potential for discomfort that often comes from consuming large amounts of proteins such as whey. This can be shown in vitro using immunoassays that quantify these peptides down to 1ppm (Rosendal A 2200-10). Whey, hydrolyzed by either Genepro Gen2 w/Prohydrolase or Pancreatin and an untreated sports digestive aid alone were examined using this method (Figure 4). The higher the bar, the more “bad” peptides are present.

Pre-measured packet of whey protein isolate without Genepro Gen2 w/Prohydrolase. The entire contents of each individual serving packet were emptied into 0.5 L of distilled water, vigorously shaken and consumed. Blood samples were collected at 0 hr (baseline, immediately prior to ingestion) 0.5 hr, 1.5 hr, 2.5 hr, 3.5 hr and 4.5 hr. The blood samples were sent to Europhins and tested for amino acids that are essential to the body and those that play a significant role in muscle protein synthesis (12 total).

Test group:

Following five days and an overnight fast, the participants returned to the laboratory for Day 2 of the study. Each participant received 500mg Genepro Gen2 w/Prohydrolase, pre-blended in 15g whey powder isolate. The entire contents of each individual serving packet containing Genepro Gen2 w/Prohydrolase were emptied into 0.5 L of distilled water, vigorously shaken and consumed. Blood samples were collected at 0 hr (baseline, immediately prior to ingestion) 0.5 hr, 1.5 hr, 2.5 hr, 3.5 hr and 4.5 hr. The blood samples were sent to Europhins and tested for twelve amino acids that are essential to the body and those that play a significant role in muscle protein synthesis. Following the study, the control data from each participant was compared to the data from the corresponding patient in either of the two test groups.

Sample collection

Whole blood samples (approximately 5 mL) were collected by a phlebotomist from multiple venous punctures, and transferred to plain Vacutainer® tubes. Serum was prepared by centrifugation and stored in 200 μL aliquots at -20°C until needed for analyses.

Analytical analyses

All serum samples were submitted to the laboratory blind to remove any analytical bias. Amino acid analyses consist of quantification of twelve individual serum amino acids for each patient at each time point. Amino acid analyses consisted of quantification of twelve individual serum amino acids for each patient over the 270 minute time period. Analyses were performed on an AA analyzer using ion exchange chromatography and a post column derivatazation with ninhydrin and UV detection.

Results

Amino acid levels increased greatly from 0 hr and the first time period taken (0.5 hr) and were fluid on return to baseline between control and test group. The shape of the curve and the quantity of leucine in the blood correlates well with leucine blood levels when taking whey hydrolysates (Tang JE 987-92). This is not a surprise since whey hydrolysate is protease treated to very small peptides, and then dried vs. whey protein that is hydrolyzed by Genepro Gen2 w/Prohydrolase at the point of consumption. This required the amino acid quantity to be calculated from area under the curve (AUC) over the entire time period and compared (270 minutes) instead of individual time points. Quantification was done versus reference standard mixtures and control mixtures of known quantities of all twelve amino acids (Sigma, St. Louis, MO). Amino acid levels are reported in μg/L. Total Serum Amino Acids (TSAA) levels were reported as the total sum of all twelve amino acids. The percent AUC was reported as the amount each amino acid contributed to the total. AUC was calculated using the KaleidaGraph software (Synergy Software, Reading, PA). Most of the samples were bell shape curves, with the lower Y limit set at the time 0 value. Results are expressed as means ± SEM.

Genepro Gen2 w/Prohydrolase had a profound increase in these amino acids. The first is the branch chain amino acids (BCAA). BCAA is a combination of essential branched chain amino acids such as leucine, isoleucine, and valine that are of special importance for bodybuilders and athletes because these amino acids are used by the body to build up protein for muscle synthesis, muscle repair, etc (Eva Blomstrand 269-73). Because BCAAs are essential amino acids, not produced by the body, they must be acquired from the diet or from nutritional supplements. BCAA is metabolized in the muscle rather than the liver; consequently the effect of these branched chain amino acids is much quicker and efficient than of any other amino acid. After BCAAs are digested, protein breaks down into individual amino acids that can either be used to build new proteins or used as energy for the body. If the diet is balanced, branched chain amino acids will be used for protein synthesis, essential for endurance athletes and strenuous workouts. BCAAs are also used to reduce fatigue in both anaerobic and endurance sports. Because of its anticatabolic properties and vital role in protein synthesis, leucine is considered to be one of the most critical BCAAs. Leucine is the strongest of the BCAAs and is responsible for the regulation of blood-sugar levels, the growth and repair of tissues in skin, bones and of course skeletal muscle. It’s a strong potentiator to Human Growth Hormone. It helps in healing wounds, regulating energy and assists in the preventing the breakdown of muscle tissue. Levels increased by 5%. Valine contributes to repair and growth of muscle tissue, as commonly attributed to BCAAs. It is not processed by the liver; rather actively taken up by muscle. It maintains the nitrogen balance and preserves the use of glucose. Levels increased by 11% . Threonine is an essential amino acid that is never manufactured within the body. Since its main sources are animal (dairy and meat) this doesn’t bode well to vegans. It’s found in heart, skeletal muscle and nerve tissue in the central nervous system. Threonine is used to form the body’s two most important binding substances, collagen and elastin. Threonine is involved in liver functioning, lipotropic functions (when combined with aspartic acid and methionine) and in the maintenance of the immune system by helping in the production of antibodies and promoting growth and activity of the thymus. Perhaps its most useful property is that it allows better absorption of other nutrients; therefore protein sources containing threonine are more bio-available than others. Levels increased by 6%. Arginine has amazing nitrogen retention ability. Nitrogen is one of the key elements in muscle protein synthesis. Some plants can absorb nitrogen, but mammals have to make do with what we make ourselves. Arginine enhances the immune system and stimulates the size and activity of the Thymus gland, which makes it a prime choice for anyone in less than optimal health, such as people recovering from injury and HIV patients. Arginine is also a precursor of very important molecules such as creatine and gamma amino butric acid (GABA, a neurotransmitter in the brain). The hormonal release properties include releasing insulin from the pancreas and a massive stimulator in the manufacture of GH (Growth Hormone) from the anterior pituitary. It increases blood flow. It also improves the health of the liver, skin and connective tissues and may lower cholesterol. But mostly it facilitates muscle mass gain while limiting fat storage, because it keeps fat alive in the system and uses it. It’s key in weight control. Levels increased by 60%, more than 2.5 times the level obtained from using whey isolate alone. Glutamine is a non-essential amino acid that is present in the body in large amounts. At some times it forms 60 percent of the total amino acid pool. Because it passes through the blood-brain barrier rather easily, it’s often called “brain-food”. In the brain it converts to glutamic acid, which is essential for brain functioning and increase GABA (gamma-amino-butyric-acid, another popular supplemented amino) needed for mental activities. It is used in synthesis of muscle tissue. It is a nontoxic nitrogen carrier. Perhaps most importantly, it balances the acid/alkaline level, so it reduces lactic acid. Levels increased by 28%. Phenylalanine, the major precursor of tyrosine, enhances learning, memory, mood and alertness. Is a major element in the production of collagen and suppresses appetite. Levels increased by 9%. Methionine is the precursor of cystine and creatine, and may increase antioxidant levels (glutathione) and reduce blood cholesterol levels. It also helps remove toxic wastes from the liver and assists in the regeneration of liver and kidney tissue. Levels increased by 6%. Total amino acid concentrations increased 60mg in the blood over the 270 minutes after ingestion of whey protein isolate with Genepro Gen2 w/Prohydrolase when compared to taking whey protein isolate alone. This correlates well with published results showing amino acids increases when taking whey hydrolysate vs. whey protein unprocessed.

Peptide discomfort is caused by the presence of specific sequences of amino acids, called epitopes, in the native protein. Epitopes are areas on the protein surface which are recognized by the immune system and identified as having between 8-16 amino acids. It is thought that a

discomfort peptide has a minimum of 14 amino acid residues. Trypsin digestion of β-lactoglobulin has shown that there are many epitopes spread over its surface. The molecular weight of a protein also determines whether it will act as a potentially discomfort material or not. Potential discomfort material consists of components with the ability to stimulate antibody production and a component with at least two antibody binding sites. The probability that a component will meet these criteria increases above a molecular weight of 3,000 Daltons(Rosendal A 2200-10). In general, the lower the molecular weight of the protein, the lower its discomfort potential. This means a reduction in the likelihood of discomfort effects. Five out of the six participants complained of discomfort when taking 50 grams of whey isolate during the control period, while no one felt any discomfort when taking 50 grams of whey with Genepro Gen2 w/Prohydrolase in the test period. These results indicate that Genepro Gen2 w/Prohydrolase is able to hydrolyze the peptides in whey that cause discomfort, consistent with the Veratox in vitro results in the laboratory.

Conclusions

The amount of amino acids found in the blood with whey and the increase with Genepro Gen2 w/Prohydrolase is consistent with the study and benefits of whey hydrolysate (Koopman106-15). Genepro Gen2 w/Prohydrolase helps break down whey protein efficiently and completely (97.8% compared to whey alone at 31.7%) so that the body can maximize the absorption of amino acids into the blood stream and reduce the exposure time to whey peptides that can cause discomfort. Consumption of Genepro Gen2 w/Prohydrolase significantly raised the level of amino acids in the blood compared with whey isolate alone. Among those amino acids are the branch chain amino acids, which have been shown to play a vital role muscle synthesis and recovery (Borsheim E648-E657). Based on the success of this clinical study, preparation for an advanced clinical to further investigate the benefits of Genepro Gen2 w/Prohydrolase/WHEY is underway.

MedFit Rx, Inc.
North Carolina – Texas – Virginia – Florida – Indiana - Maryland
300 Ashville Ave Suite 230
Cary, NC

Khmara Laboratory
Chicago, IL
Completed: April 2, 2016

Correspondence to:

Dr. Bria M. Parks, PhD
COO
Musclegen Research, Inc.
PO Box 607
Willow Springs, NC 27592

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