Hydrolysates. Pre-digested proteins for performance nutrition

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1 Hydrolysates Pre-digested proteins for performance nutrition

2 Introduction Proteins are an essential part of a diet. Protein helps to maintain muscle mass and have an important role to regulate processes in the body. In performance nutrition, protein is widely accepted to help to recover your muscle mass. For both endurance and resistance exercise athletes it is recommended to ingest more protein then the recommended 0.8g/kg/bw for an average adult (see table 1). Digestion in the body Proteins are long chains of amino acids linked through peptide bonds (see figure 2). When an intact protein arrives in the stomach, enzymes (like pepsin) start to hydrolyze these peptide bonds. This results in a peptide mixture from bigger peptides (multiple amino acids linked together) to small peptides (a few amino acids linked together). Finally in the intestine, single amino acids, di- and tripeptides are absorbed into the bloodstream and transported where the body needs them (see figure 1). Estimated protein requirements Adults with sedentary lifestyle 0.8 Recreational moderate endurance athlete Elite male endurance athlete 1.6 Resistance (steady state) athlete 1.2 Table 1: Protein requirements for different athletes (1) g/kg/body weight Resistance (early trainings) athlete Enzymes Pepsin Trypsin Chymotrypsin carboxypeptidases Carboxypeptidases Proteins Large polypeptides Small polypeptides Amino acids Di- & tripeptides For the active body it is important to have the nutrients when the body needs it most. Appropriately timed protein intake is an important component of an overall exercise training program, essential for proper recovery, immune function and maintenance of lean body mass. The overall advice is to consume protein immediately after exercise to maximize muscle protein synthesis. Proteins composed of different amino acids. In total there are 20 different amino acids. 9 amino acids are defined as essential amino acids; your body cannot synthesize themselves and have to be provided with nutrition. Conditionally essential amino acids (such as arginine and histidine) have to be consumed in the diet at some stages of growth or by certain circumstances such as medical condition or genetics. Every protein source has their own unique amino acid profile. Milk protein (casein and whey) are considered as complete proteins, which means all essential amino acids are sufficient available. Fig. 1: llustration of enzymatic protein digestion in digestive tract. Today we have different ways to get amino acids into the bloodstream: Our daily nutrition Protein rich supplements (which can be supplied/ingested with shakes, bars, powder applications, tablets etc.): - Intact protein supplements - Hydrolyzed (pre-digested) protein supplements - Free form amino acids What is a protein hydrolysate? Protein hydrolysates are produced from intact protein sources by the addition of enzymes. Depending on the type of enzyme(s) used and processing conditions chosen, it results in different patterns of peptides and amino acids. In figure 2 an overview of different peptide bound chains that can occur in a protein hydrolysate is given. Every protein hydrolysate has their unique mixture of peptides. Intact protein Polypeptides Tripeptides Dipeptides (single) Amino Acids Fig. 2: Appearance of different peptide bound amino acids.

3 Important parameters of hydrolysis characteristics Each protein hydrolysate is a complex mixture of peptides of different chain length together with free amino acids. A protein hydrolysate can be defined by the degree of hydrolysis (DH), but DH does not tell us the whole story, as two protein hydrolysates made by different methods may have a similar degree of hydrolysis even though their absorption kinetics are likely quite different. Consequently, all protein hydrolysates are certainly not created equal. Peptide 1 Molecular weight distribution (MWD) The molecular weight distribution (MWD) profile reflects the functional and nutritional properties of the protein hydrolysates. Using this method, peptides are separated according to their size and shape, which is related to their molecular weight. Free Amino Acids (FAA) The free amino acid content is the amount of amino acids that is not peptide bound. Typically, the result is expressed as the sum of the individual values: % total free amino acid content. It is also possible to determine which amino acid is a free amino acid. Calculation of the degree of hydrolysis The degree of hydrolysis (DH) is defined as the percentage of the number of peptide bonds in a protein which has been cleaved during hydrolysis. Peptide 2 Fig. 3: Schematic presentation of two different poly peptides cleaved by enzymes (red stripes). The amount of cuts both peptides have is the same but the characteristics after cutting is different. Peptide 2 has more di- and tripeptides compared with peptide 1. There is a range of parameters to describe the characteristics of protein hydrolysates: Amino Nitrogen Total Nitrogen Ratio (AN-TN) Molecular Weight Distribution (MWD) Free Amino Acids (FAA) Degree of Hydrolysis (DH) There are more parameters to describe the characteristics of protein hydrolysates. They are not further described in this brochure, knowledge on these and other parameters is available and we are willing to share this. Amino Nitrogen (AN) and Total Nitrogen (TN) Amino Nitrogen represents the amount of all free amino groups in a hydrolysate or protein which include those in free amino acids, the N-termini of peptides and proteins and the side chain amino groups in lysine. Total Nitrogen content is quantitatively determined by methods such as Kjeldahl or Dumas and includes all nitrogen in the product, including non-protein nitrogen and nitrogen incorporated in amino acid side chains.

4 Nutritional benefits of protein hydrolysates Protein hydrolysates are typically manufactured by enzymatically treating proteins, similar to the human digestive process (see also figure 1). Therefore the protein hydrolysates can almost immediately be absorbed and are likely to be digested faster into the bloodstream (2). Di- and tripeptides are absorbed faster than free amino acids Protein hydrolysates containing mostly di- and tripeptides are absorbed more rapidly than free form amino acids and much more rapidly than intact proteins (3). In the intestine different carriers for proteins exist to transport them to the bloodstream (figure 4). In the brush border membrane there is more capacity to transport di- and tripeptides compared with single amino acids (3). The chain length of the peptides has an important influence on the absorption rate of the peptides into the bloodstream (4). Therefore the body is better equipped to transport di- and tripeptides compared with single amino acids (5). Proteins Peptides amino acids Carrier di- and tripeptides Carrier single amino acids di- and tripeptides Benefits di- and tripeptides Small intestine Bloodstream Di- and tripeptides are absorbed faster than intact proteins. The body is better equipped to transport di- and tripeptides. Whey protein hydrolysates are digested fast and have a positive effect on muscle protein synthesis Tang et al (6) showed whey protein hydrolysates are digested fast, but it has also impact on the muscle protein synthesis. Brush border membrane Fig. 5: There are more carriers for di- and tripeptides than single amino acids. This study shows whey hydrolysate stimulates muscle protein synthesis to a greater degree than intact soy or casein protein after resistance exercise (figure 6). Muscle protein synthesis is essential to the body s ongoing growth, repair and maintenance of the skeletal muscle groups. Both exercise and proteins are signals to activate muscle protein synthesis (7).. Di-and tripeptides cotransport with H + H + Na + Amino acids cotransport with Na + Small peptides are carried intact across the cell by transcytosis * # Rest Ex Peptidases FSR (%.H 1) * * * H Na Whery Casein Soy Blood To the liver Fig. 4: Carriers for peptides. Fig. 6: Mixed muscle protein fractional synthetic rate (FSR) after ingestion of whey hydrolysate, casein, or soy protein at rest and after resistance exercise. *Significantly different from casein for same condition (P < 0.01). # Significantly different from soy for same condition (P < 0.05). All values are means ±. Figure adapted from Tang et al (6).

5 To increase muscle mass and maximize muscle recovery besides intensive exercise, nutritional strategies are very important. Studies (8,9) have demonstrated that protein ingestion can increase post exercise muscle protein synthesis and net protein balance. Benefits whey protein Whey protein is a fast protein. Whey protein has a positive effect on muscle protein synthesis. Whey protein hydrolysates on muscle glycogen responses and insulin To improve endurance-exercise performance dietary interventions generally aim to maximize glycogen availability. Muscle glycogen is the most important fuel source during prolonged moderate to highintensity exercise (10). There is a direct relationship between fatigue and muscle glycogen depletion (11). To maximize glycogen availability many carbohydrate-based drinks have been developed. Skeletal muscle glycogen synthesis is a relatively slow process and complete restoration of the glycogen stores after exercise generally takes up to 24h (12). Numerous studies have been described to find the optimum amount of carbohydrates. Hydrolysates on glycogen depletion Excellent amino acid profile Whey has a higher leucine content compared with other sources such as casein or soy protein. Leucine has been identified as the main nutritional signal responsible for stimulating postprandial muscle protein accretion. Benefits leucine Leucine is a signal amino acid, responsible for stimulating muscle protein synthesis. Whey derived from fresh milk has a higher leucine content compared with whey from cheese. This can also contribute to the proposed greater anabolic properties of whey than casein, because leucine has been identified as the main nutritional signal responsible for stimulating postprandial muscle protein accretion (15). Koopman et al (16) found ingestion of protein with additional leucine and carbohydrates improves whole body protein balance during recovery compared with beverages lower in leucine. A higher leucine content is likely an effective strategy to increase anabolism after resistance exercise. Whey protein has a high leucine content, but the production of whey has also impact on the amino acid profile. Whey protein made from cheese whey has a lower leucine content compared with whey derived from fresh milk (own data). Glycogen depletion is associated with fatigue during and after exercise. Protein hydrolysates in combination with carbohydrates are considered to accelerate muscle glycogen synthesis. Insulin seems to be one of the key hormones in this process. Whey protein hydrolysate activates insulin response and glycogen synthesis, thereby attenuating exercise-induced glycogen depletion. But not only carbohydrates are important to optimize muscle glycogen stores. Observations by various research groups show that coingestion of protein and protein hydrolysates with carbohydrates further accelerates postexercise muscle glycogen synthesis when compared with ingestion of carbohydrate only (13). The mechanism behind the effectiveness of this strategy is quite simple. Insulin is a hormone that regulates many processes in the body. One of the tasks of insulin is to stimulate glucose utilization through the activation of glucose transport, which is generally considered to be the major factor to determine the rate of glycogen synthesis. Ingestion of dietary protein is known to induce insulin and glucagon secretion (14) Therefore, an increase in postexercise insulin response, after the ingestion of protein (and amino acids) in combination with carbohydrates, has been suggested to accelerate muscle glycogen synthesis (10).

6 A selection of our promotional support documentation range Application suggestion 3 stage protein bar Application suggestion Endurance protein drink Application suggestion Recovery on the go tablets Excellion EM 9, Nutriwhey 800 F & Hyvital Whey 8022 A tasteful high protein bar, to be consumed after exercise, formulated with three types of protein, each with different digestion rate; Excellion EM 9 Calcium Caseinate, slow digestion, Nutriwhey 800 F, whey protein concentrate, demonstrated as a fast digested protein (1) and Hyvital Whey 8022 whey protein hydrolysate, which is likely immediately reach the bloodstream after ingestion. (2) Benefi ts of Excellion EM 9 Prolonged release of amino acids into the blood stream Calcium Caseinate is a good source of bioavailable calcium Contributes to decrease stickiness in a high protein bar Benefi ts of Nutriwhey 800 F Hyvital Whey 8016 A protein-carbohydrate drink, to be consumed during resistance exercise, for maximized glycogen availability and optimized post exercise recovery. This drink contains Hyvital Whey 8016, a Whey protein hydrolysate, with a unique composition, making it ideal for protein supply during intense exercise. Benefi ts of Hyvital Whey 8016 Fast release of amino acids into the blood stream High in di- and tripeptides that are better absorbed compared to single amino acids High Leucine content Hyvital Whey 80 chilsonated Our ingredients are developed to meet ever increasing consumer needs. There are essentially three different ways to take protein; intact protein, hydrolyzed or pre digested protein and amino acid mixtures. In this application we give you an excellent example of a hydrolyzed or pre-digested protein, based on whey protein. Benefi ts of Hyvital Whey 80 chilsonated Excellent tabletting properties For fast absorbing whey peptides, aiding fast recovery after exercise Convenience in delivering high content natural BCAA s There are several ways to ingest the right Whey protein hydrolysates can almost amount of nutrients after exercise, mostly on immediately be absorbed and are likely to powder form, as a ready to drink or as a bar. be digested fast. (3) Bars are becoming more and more popular Whey protein has been suggested to among consumers. For the producers, the maximize muscle protein synthesis because challenge is to formulate products with the of fast absorption and a high amount of right amount and ratio of available nutrients leucine. (4) while creating a tasteful treat to be Caseinate has a more prolonged release into consumed after exercise. the bloodstream and is therefore also named With this recipe, we created a 3 stage bar as the slow protein. (5) with three different sources of protein, Besides the prolonged protein release this having the advantages of different three protein blend ensures optimum bar absorption rates in the body. mouthfeel and structure. In three stages it is possible to benefi t This recipe is only one suggestion of many. maximal of the different absorption rates DMV can support you tailoring protein bar of various proteins, giving the athlete a recipes towards your desired structure and sustainable release of amino acids over time. nutritional properties. Fast release of amino acids into the blood stream Contributes to smoothness and homogeneity in a high protein bar Benefi ts of Hyvital Whey 8022 Very fast release of amino acids into the blood stream High in di- and tripeptides Contributes to bar softening allowing higher protein formulations Is there a need for protein ingestion during exercise? It is known that protein ingestion following exercise augments muscle protein mass by further increasing muscle protein synthesis rates during resistance or endurance exercise. (1) The timing of protein ingestion has been identifi ed as another key factor for the muscle protein anabolism. (2) As a consequence, it is now generally advised to ingest 20 grams of a high-quality dietary protein after exercise. However, recent scientifi c research suggests protein can be consumed prior to and/or during exercise to further stimulate postexercise muscle build up. (3, 4, 5) This dietary strategy might facilitate muscle reconditioning and improve training effi cacy, especially for those spending many hours in training. (6) During prolonged exercise (>5 hours) a mix of protein and carbohydrates improves whole-body protein balance. (4) Protein hydrolysates are normally suggested during exercise because of their fast absorption. Hyvital Whey 8016 contains with high amount of natural available Leucine and di- and tripeptides, which are more rapidly absorbed than free form amino acids and much more rapidly than intact proteins. (7) This recipe is only one suggestion of many. DMV can support you with tailored protein drink recipes for specifi c structure and nutritional properties. The primary benefi t of protein hydrolysates digestive process, which results in a mixture is that they are absorbed faster compared to of peptides and amino acids. Our Hyvital intact proteins. Our range of Hyvital protein hydrolysates are available from light to hydrolysates are either prepared from casein, extensively hydrolysed, suitable for all kind of whey, wheat or soy. Protein hydrolysates applications either as sole protein source or are typically manufactured by enzymatic in addition to intact proteins. cleavage of proteins, similar to the human References 1. Burke, Deakin. Clinical sports nutrition. Fourth edition McGraw-Hill companies. 2. Manninen AH. Protein Hydrolysates in sports and exercise: a brief review. J Sport Sc and Med (2004) 3, Manninen H., Protein Hydrolysates in sports nutrition. Nutrition and Metabolism 2009, 6: Grimble GK, Keohane PP, Higgins BE, Kaminski MV & Silk DB (1986). Effect of peptide chain length on amino acid and nitrogen absorption from two lactalbumin hydrolysates in the normal human jejunum. Clin Sci (Lond)71, Daniel H (2004) Molecular and integrative physiology of intestinal peptide transport. Annu Rev Physiol 66, Tang, Moore, Kujbida, Tranopolsky, Phillips. Ingestion of whey hydrolysate, casein or soy protein isolate: effects on mixed muscle protein synthesis at rest and following resistance exercise in young men. J Appl Physiol 107: , Cermak NM., Res PT, Groot LC, Saris & van Loon. Protein supplementation augments the adaptive response of skeletal muscle to resistance-type exercise training: a metaanalysis. Am J Clin Nutr 2012; 96: Rasmussen, B.B., Tiption, K.D., Miller, S.L. Wolf, S.E.& Wolfe, R.R. (2000) An oral essential amino acid-carbohydrate supplement enhances muscle protein anabolism after resistance exercise. J. Appl Physiol.88: Tipton,K.D., Ferrando, A.A. Phillips, S.M. Doyle, D.J & Wolfe, R.R. (1999) Postexercise net protein synthesis in human muscle from orally administered amoino acids. Am.J.Physiol. 276:E628-E Van Loon, LJC., WHM Saris., Verhagen H., and Wagenmakers. Plasma insulin responses following the ingestion of different amino acid/protein carbohydrate mixtures. Am. J.Clin Nutr. 72:96-105, Karlsson J, Saltin B. Diet, muscle glycogen and endurance performance. J Appl Physiol 1971;71: Burke, L.M., G.R. Collier, S.K. Beasley, et al. Effect of coingestion of fat and protein with carbohydrate feedings on muscle glycogen storage. J.Appl. Physiol.78: , Van Loon, LJC., Kruijshoop, H. Verhagen, WH. Saris and AJM Wagenmakers. Ingestion of protein hydrolysate and amino acid-carbohydrate mixtures increases post-exercise plasma insulin response in humans. J Nutr 130: , Hall WL, Millward DJ, Long SJ & Morgan LM (2003) Casein and whey exert different effects on plasma amino acid profiles, gastroinstestinal hormone secretion and appitie. Br J Nutr 89, Pennings, Boirie, Senden, Gijsen, Kuipers, van Loon., Whey protein stimulates postprandial muscle protein accretion more effectively than do casein and casein hydrolysate in older men. Am J Clin Nutr 2011; 93: Koopman R, Wagenmakers AJ, Manders, Zorenc JM, Gorselink M, Keizer HA, van Loon LJ. Combined ingestion of protein and free leucine with carbohydrate increases postexercise muscle protein synthesis in vivo in male subjects. Am J Physiol Endorcrinol Metab. 2005; 288(4):

7 DMV Head Office NCB-Laan 80, P.O. Box 13, 5460 BA Veghel, The Netherlands T F info@dmv.nl Latin America Office Rua dos Canarios 65 Vinhedo, SP Brasil T renata.soliva@frieslandcampina.com North America Office 1285 Rudy Street Onalaska, WI 54650, USA T F bill.brickson@frieslandcampina.com Copyright DMV, The Netherlands, April 14 Asia Pacific Office 3 Temasek Avanue # Centennial Tower Singapore T F florian.middelhuis@frieslandcampina.com China Office Rm 1573, 15FI NCI Tower 12A Jianguomenwai Ave. Chaoyang Dist Beijing, P.R.C. T F martijn.jager@frieslandcampina.com Warranty Questions? Technical support? Advice? Call the Hydrolysates experts The details given in this document are merely intended for information purposes and are in no way legally binding. Consequently we accept no responsibility in the broadest sense of the word for damage that may result from applications based upon this information. Furthermore, this information does not constitute permission to infringe patent and licence rights. DMV is a brand of

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