Team Muscle Intelligence 2016

PROTEIN
HYDROLYSATES
A SYSTEMATIC REVIEW AND COMPARISON WITH INTACT
PROTEIN OR AMINO ACID SUPPLEMENTATION
Moritz Werner, Stefanie Macherhammer

For decades, research on amino acid supplementation, in particular essential and branched chain
amino acids, as a fast acting Protein source and their effects on body composition and skeletal muscle
growth remains extremely controversial, failing to consistently show a clear cut benefit over plain protein
supplementation or on top of a high nutritional protein intake. (1-10)
While the authors of Team Muscle Intelligence involved in this systematic review share the stance of the
Journal of the International Society of Sports Nutrition (3) and believe that there are certain situations where
the application of free form amino acid supplements can be useful, we want to focus in our scientific review
on another fast acting protein source, that in numerous lines of research, with much more consistency and
less controversy shows to be superior in terms of 1) absorption kinetics, 2) anabolic signaling, resulting
in more glycogen and muscle protein accretion 3) recovery, performance and muscle adaptions to both
normal non-hydrolyzed protein and amino acid supplementation:
Protein hydrolysates (PHY), in particular whey- and casein-hydrolysates (WHY) (CHY) .

1. Absorption Kinetics of protein hydrolysates

One of the major reasons for an athlete to use amino acid supplements is to have a more rapid and
available protein source, resulting in a faster decrease in muscle protein breakdown and increase in protein
synthesis around his training session to recover more quickly and adapt faster to the training stimulus.
While the importance of protein timing around training sessions has gotten questioned lately, (11) more and
more lines of research seem to confirm, that with growing training age and experience it is a practice that is
becoming increasingly important for the advanced trainee. (12-13)
It intuitively makes sense for the athlete to use the fastest absorbing and most readily available protein
source before, after or during his workout and that free form amino acids seem to be a favorable choice
over whole protein supplementation for this purpose.
Nevertheless protein hydrolysates, having a high di- and tripeptide content seem to have favorable
absorbing kinetics, including better and faster uptake by skeletal muscle, earlier and stronger
hyperaminoacidemia hyperisulinemia, anabolic signaling and protein synthesis over both non hydrolyzed
proteins and free form amino acids. (4-8, 14-18, 25,26)
This seems to be due to the independent uptake pathway of peptide transporters, allowing a more efficient
uptake of amino acids, in their di- and tripeptide bound forms. (4-8, 14-18, 25,26)
In this first section we conclude, that protein hydrolysates with a high di- and tripeptide content, would be
preferably used over free form amino acids, whenever protein timing and fast availability seems to be of
major importance.

2. Anabolic Signaling
In this section we want to focus on the impact of protein hydrolysates on several different anabolic
signaling on several different anabolic signals and signaling pathways such as insulin, mTor, AKT, GLUT4translocation and the resulting increased glucose uptake by skeletal muscle and nitrogen retention, leading
to improved glycogen replenishment, restored protein balance and super-compensation of both.

2.1 Protein = More than the sum of its parts

It is often hypothesized that amino acid supplementation will provide a favorable response over protein
ingestion, because of greater peaks in blood plasma amino levels, interestingly most studies comparing
protein supplements with a mixture of identical free form amino acid mixtures will deny this thesis: Showing
greater anabolic signaling and net protein balance for the protein supplement. (4,5,6,8) This leads most
researchers to the conclusion that the anabolism of protein goes beyond its constituent amino acid
content, providing additional benefit in its unique peptide structures; including growth factors like IGF-1,
immune factors like immune-globulin A and B, as well as certain functional, branched chain amino acid
containing di- and tripeptides (4,6,23,24,25,27,29,36-39)
Since protein hydrolysates are only partially hydrolyzed, still containing most of these small and long chain
functional peptides, the authors believe that they provide the best of both worlds; having a more rapid
availability than free form amino acids while still providing all the above described benefits of whole non
hydrolyzed protein. Which is supported by Kanda et. al. (4) using a protein hydrolysate for their comparison
with the amino acid mixtures in their work.

2.2 Insulin secretion and signaling

Protein hydrolysates are highly insulinotropic, having a substantial impact on insulin secretion on their own
and when co-ingested with carbohydrates, being also significantly more effective secretogoges than intact
proteins and free form amino acids (4,6,8,19,23,25,29,44). This could be due to specific peptides found in
dairy protein hydrolysates such as certain branched chain amino acid peptides like the Leucyl-Isoleucine
peptide. (25,29)
Several studies found that co-ingesting protein hydrolysate can lead to increases of up to 299%
over glucose only, in insulins area under the curve on plasma levels. With CHY showing the highest
insulinotropic effect measured in all studies. (19-22,25,29,31,33-35,44)
While it is established that insulin in supra-physiological levels induced by exogenous administration can
promote anabolic advantages for the growth of skeletal muscle and lean body mass, which is one of the
reasons for bodybuilders to abuse this drug (32), it is getting frequently questioned in the literature whether
fluctuations of insulin from carbohydrate consumption within normal physiological ranges would exceed
the anabolic threshold needed for insulin to further positively influence net proteins balance when ample
protein is provided. The authors of this review are convinced that such high levels of insulins AUC as
shown in (21-22) are indeed high enough to exceed the threshold needed for insulin to provide additional
anabolic enhancement over ample protein ingestion alone, which seems to be supported of the data by
Van Loon (2000) demonstrating that such an increase in plasma insulin concentration, during conditions of
hyperaminoacidimea, further increases net protein balance (34,35). As well as Calbet and MacLean (2002),
concluding, that the association of high levels of plasma amino acids and insulin might explain a superiority
of peptide hydrolysates over whole proteins in promoting better nitrogen utilization, especially when
administered in combination with glucose (33).

2.3 AKT, MTor, AMPK, -signaling and

GLUT4-translocation.
Alongside with a bigger impact on hyperinsulinemia and hyperaminoacidimea peptide based proteins lead
to a significant increase in anabolic signaling genes and pathways like mTor, AKT AMPK, S6K1 and
4E-BP1 (4,6,8,29,30) over amino acid mixtures. Morato et al. (2013) show, that specifically the
insulinotropic Leu-Ileu peptide found in protein hydrolysates seems to have bigger impact on AKT, AMPK
signaling and insulin dependent GLUT4 translocation than leucine itself (29). Also Kanda and his researches
concluded, that The WPH the present study contained active components that are superior to leucine
itself for increasing muscle protein synthesis (Kanda. 2013). (4,27)

2.4 Glycogen replenishment

As a result of enhancing the previously described anabolic signaling pathways, peptide based proteins
significantly increase glucose uptake and glycogen storage in skeletal muscle via activation of the GLUT4
nutrient transporters, providing substantial benefit over intact proteins and amino acid mixtures. This also
seems to be in context with specific branched chain amino acid peptides found in protein hydrolysates.
(23-30)
The amino acid compositions of the two diets used in this study contained equal amounts of BCAA and
leucine; however, muscle glycogen accumulation varied between diets. This result strongly suggests that
not only the BCAA content but also the molecular form of BCAA found in the protein source might be
important for muscle glycogen storage. (Kanda. 2012) (27)
Kanda et al. do then refer to a previous study, in which his group had been able to demonstrate that
BCAA-containing peptides in WPH, which have been shown to be markedly elevated (meaning they are not
digested) after the consumption of whey protein hydrolysates (Morifuji. 2010) (25) in a follow up study on
human beings, they actually have the ability to stimulate the rate of glucose uptake in vitro (Morifuji. 2009)
(23)
This is making protein hydrolysates not only a useful supplement to enhance post workout recovery and
anabolic adaptions, but also a very effective tool to enhance carbohydrate loading protocols resulting in
increased glucose uptake and retention by muscle and leaving less substrate that could potentially stored
in adipose body fat tissue.

3. Recovery, performance and muscle growth

While all data about the improved anabolic signaling, nitrogen retention, muscle protein balance and glycogen
replenishment in a super-compensatory manner is clearly impressive and promising, we want to sum this
review up with real world data and results that show the effectiveness of protein hydrolysates to improve
recovery from hard training sessions and restore, or even improve performance as quickly as possible.
In a Brazilian study Pablo Christiano B. Lollo et al. investigated the effects the provision of whey protein
(WP), hydrolyzed whey protein (WPH), or a non-protein placebo maltodextrin, (MALTO) would exert on
selected performance and muscle damage related parameters, finding that hydrolyzed protein compared
with its intact relative significantly decreased muscle damage indicators, creatine kinase (-42%) and lactate
dehydrogenase (-30%). The minimal changes in the whey protein group, on the other hand, were not
superior to the maltodextrin control. (43)
In a 2012 study from Journal of the International Society of Sports Nutrition Buckley et al wanted to
determine whether hydrolyzed whey protein speeds recovery following isometric exercise, males performed
100 maximal eccentric contractions of their knee extensors and then consumed either 24g of hydrolyzed
whey protein or intact whey protein. Peak isometric torque recovered fully in 6 hours for individuals given
the whey protein hydrolysate whereas it remained suppressed in the group given intact whey protein. (42)
In another paper from Cribb et al. 2006, recreational male bodybuilders supplemented their normal diet
with either hydrolyzed whey protein isolate or a non hydrolyzed protein during a 10 week supervised
resistance training program. The group supplemented with the hydrolysate achieved substantially greater
gains in muscle strength and lean body mass compared to the non-hydrolyzed control group (41)
Therefore, whey protein hydrolysates offer ergogenic benefits when consumed during and after exercise
and seem to improve recovery, performance and muscle growth in athletes.

Conclusion
In this review we have shown that protein hydrolysates yield substantial advantage over intact proteins and
amino acid mixtures in terms of 1) Absorption kinetics 2) Anabolic signaling, nitrogen balance and glycogen
replenishment 3) enhancing recovery, performance and anabolic adaptions in athletes.
We want to point out that several studies in our references have been looking specifically at the hydrolyzed
dairy derived proteins whey and casein, with high di- and tripeptide content, which both specifically have
great biological value and are rich in branched chain amino acid containing peptides as well as growth
and immune factors that seem to be one of the major key component, responsible for their superiority in
anabolic signaling.
Since different studies show different advantages of WHY and CHY over each other, CSY showing the
highest insulinotropic response shown in all performed studies (19-22,33-35) and enhancing performance
relevant parameters over WHY (45,46), but whey showing significantly better protein synthesis response
post workout in another paper (44);
Team Muscle Intelligence is recommending a 1:1 blend of both at approx 0,4-0,5 g per kg of body weight
to their athletes approx. 30 minutes post workout to maximize anabolism and recovery alongside with high
glycemic, high molecular weight carbohydrate.

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