Pasteurized Polypeptide Powder

The Science Behind this Unique Advanced Protein Product

Roger W. Law, Chief Technology Officer

Asta Ltd.

Introduction

Proteins are the true workhorses of the body, carrying out most of the chemical processes and making up the majority of cellular structures. They are required for the structure, function, and regulation of the body’s cells, tissues, and organs. Proteins are fundamental components of all living cells and include many substances, such as enzymes, hormones, and antibodies, muscles, skin, bones and the body as a whole. Each protein has unique functions. They are essential in the diet of humans and animals for the growth and repair of tissue, and are obtained from foods consumed such as meat, fish, eggs, etc.

Proteins are essential molecules woven into the fabric of life. They are defined as any of various naturally occurring extremely complex molecules that consist of amino acid residues joined by peptide bonds and include many essential biological compounds. Amino acids are the building blocks of proteins. When amino acids link together they form a peptide; when two amino acids link together they form a di-peptide, and three linking together form a tri-peptide. A single peptide molecule may be only a few thousand Daltons; a term used to express the atomic mass of a molecule. Proteins can be as small as 35,000 Daltons to over a million Daltons. The term polypeptide applies when more than three peptides link together to form a chain; chains of polypeptides create proteins.

A new advanced protein product, termed Pasteurized Polypeptide Powder (PPP), is about to be introduced into the market. This advanced protein product exhibits unique characteristics and superior performance benefits. More specifically it exhibits extremely high bioavailability, is offered in a powdered form, possesses an unusually long shelf life, is hypoallergenic, and has a high degree of palatability. Scientific research has confirmed these superior properties. The combination of these characteristics plus its low cost of manufacturing makes PPP the first new advanced protein product in decades derived from animal sources having applicability throughout the entire human food chain. The human food chain consists of aquaculture and animal feed, as well as direct human consumption.

Asta Ltd., the company that will bring PPP to the market, is a Northwest U.S. based food biotechnology company that owns a proprietary process that transforms a stream of underutilized and non-marketable protein sources into valuable pasteurized “advanced protein product powder.” The company’s biotechnology is an innovative continuous enzymatic hydrolysis process. The significance of this process is that it only takes one hour from raw material input to finished product, it can be operated continuously (24X7), and it takes relatively few operators to function proficiently. PPP is expected to be produced with a low cost of goods sold since raw material costs are forecasted to be low and the processing technology is very efficient. Clean, pasteurized water is the only by-product that is returned to the environment from the process.

Marine species, derived from seafood processing food grade discharge streams, are the raw materials of choice because they are available, they are animal protein, and they possess other high value nutrients. Asta’s process converts marine species into four noteworthy products These are:

   a.) PPP

   b.) omega-3 oils

   c.) calcium phosphate

   d.) chondroitin

It is believed that this new advanced protein product will compete favorably with fishmeal, soy and whey protein concentrates an isolates.  Fishmeal is the protein of choice in aquaculture and in some areas as animal feed. Currently there are over 5 million metric tonnes of fishmeal used yearly. Soy and whey protein isolates are expensive to produce. Fishmeal is not used for human consumption since it is not palatable and soy and whey concentrates/isolates are too expensive and thus, are limited to human consumption only. It is believed that PPP can create an entirely new global market as an advanced protein product ingredient applicable for the entire human food chain.

Proteins and Protein Digestibility

Proteins are very large and complex molecules made of linked chains of polypeptides. In the digestive process they are broken down by protease enzymes into smaller and smaller chains; to polypeptides, tri-peptides, di-peptides, peptides, and finally to amino acids. In the small intestine (see Figure 1) these reduced molecules are absorbed through tiny-finger like projections called villi, that serve as the intestinal-blood stream barrier, where small molecules enter the blood stream as amino acids, di-and tri-peptides. These are used to build myriads of specific proteins for all kinds of life sustaining purposes. Research has demonstrated that a well-balanced mixture of essential and nonessential amino acids is a requirement for proper health and growth.^[1]

However, some proteins are not completely broken down (digested) and therefore are too large to be absorbed through the intestinal wall. These large protein molecules are passed through the digestive track, are eliminated and not used. Considering feed formulations it is important that protein comes in a digestible form: thus reducing  the cost of production and the volume of animal waste generated.

PPP Unique Characteristics

PPP derives its unique characteristics from two primary variables: 1) food quality animal protein raw materials, and 2) innovative processing technology.

Only animal protein contains all the essential amino acids necessary for human nutrition, whereas vegetable protein typically lacks one or two essential amino acids. Asta’s innovative processing technology transforms perishable animal protein into shelf-stable, pasteurized, short chain polypeptide powder for ease of storage, transport, and use elsewhere as an ingredient. Additionally, depending on the raw material, PPP may be a source of polyunsaturated fatty acids, essential omega-3 oils which are widely used as nutraceuticals, and contain vital energy dense calories.

The short chain polypeptide powder is the result of Asta’s innovative enzymatic partial hydrolysis process. This partial hydrolysis imparts important characteristics to the PPP such as:

*Feed trial results: Digestibility, protein efficiency ratios, conversion rates, and palatability along with other relevant related criteria were documented in a series of feed trials on salmon and mink in the early 1990s. The results of these non-published studies are available upon request.

Scientific Study Overview

An appropriate place to begin in the overview of scientific studies as they relate to animal protein enzymatic hydrolysis and hydrolysates is to quickly review the basics.

1.  Enzymatically hydrolysed animal protein hydrolysates are complete proteins; containing all the essential amino acids the body can not synthesize.

2.  The molecular weight of hydrolysate components determines physical properties and characteristics and therefore, is critically important to control, monitor and document.

3.  The components of animal protein enzymatic hydrolysis are fundamental nutrients for life.

The body of scientific knowledge related to protein hydrolysates is voluminous and overwhelmingly favorable. From feeding trials in aquaculture,^{[2],[3],[4],[5]}companion pets food studies,^[6],[7]and in human diet studies,^[8],[9],[10] protein hydrolysates are nutritionally equivalent to whole or intact proteins, and exhibit superior properties and excellent test results. As an example, in aquaculture feed studies there is ample evidence of superior results on fish fed diets based on protein hydrolysates vs. diets of fishmeal formulations:

• Decrease in larval skeletal malformations by 91.4%[11]
• Growth and survival rates were significantly better[12]
• Significantly better conversion rates[13]
• 99% protein digestibility in salmon[14]
• Significantly better growth rates[15]

The results of these feed trials clearly demonstrate that a diet of PPP will increase profitability through more efficient utilization of feed and increased survival rates of fish.  By inference this means that there will be less waste (both animal waste and unused feed waste) to pollute aquaculture ponds and a significant increase in survival rates translates into a greater number of fish harvested. Less pollution in the pond has important benefits since less waste translates into less cost of cleaning ponds, a lower amount of bacteria formation, and a corresponding lower amount of fish population diseases and ultimately a lower cost of anti-bacteria and anti-fungicide pharmacological agents being introduced into the pond.

Considering companion pet studies, hydrolyzed proteins increase palatability^[16],[17] and can reduce protein allergenicity:^[18] Allergenicity has been shown to be significantly reduced when polypeptide molecular chain lengths are less than 20 kDaltons. (Note: PPP is comprised of polypeptide chain lengths of less than 20 kDaltons.)

For humans, in addition to hypoallergenic properties,^[19] protein hydrolysates are commonplace in foods for sports nutrition, infants, and patients with digestive tract complications.^[20] Because of reparative properties linked with reduced molecular weight, protein hydrolysates may prevent and treat internal injuries and damage, associated with the bowel.^[21] Therefore, hydrolysates are widely used –

Despite the tremendous amount of information available on the topic of protein hydrolysates, it is intriguing and appropriate to note the dearth of research available on polypeptide protein hydrolysates. And there is even less discussion regarding polypeptide protein hydrolysis process parameters, enzymes used, molecular weights, and the variety of starting raw materials involved. These considerations are all very significant in properly characterizing and standardizing the attributes for a true polypeptide product entry into the market place.

In the scientific community there are three classifications^[22] of protein hydrolysates. These are:

I). Extensively Hydrolysed Protein – defined by the American Academy of Pediatrics as amino acids and peptides having an average molecular weight of < 1,500 daltons^[23],[24]

II).  Partially Hydrolysed Protein – defined as smaller than an intact or whole protein, and larger than 1,500 daltons

III).  Free Amino Acid-based — no protein chains; all basic amino acids

Asta believes that these classifications are too broad since they do not adequately take into account unique characteristics of polypeptides from 1.5K daltons to 20K daltons. Over time, as PPP achieves market penetration Asta will lobby the appropriate scientific panels to adopt a greater degree of stratification to properly reflect different characteristics within narrower dalton ranges.

Other Products and Their Significances

Although this paper’s emphasis is to introduce to the reader PPP and to highlight its potential importance to the world as an advanced protein product, it is likewise important to note that Asta’s process using its targeted raw material will produce other high value added products. These are

1. Omega-3 oils. Asta’s omega-3 oils are food supplement quality and have little if any oxidation. For reasons still not completely understood, Asta’s Omega-3 oils do not seem to oxidize over time, therefore they have an extraordinary shelf life and no rancid odor. In humans and companion pets, daily ingestion of Omega-3 oils have been linked to improvement in many common maladies. The two most significant claims associated with daily doses of omega-3 oils are improved cardiac function and improved mental acuity.
2. Calcium phosphate. Asta’s Calcium is derived from fish bones, and possesses the same relative ratio of calcium to phosphorus as human bones. Once ingested, calcium is efficiently utilized in strengthening human bones.^[25]
3. Chondroitin. If sharks, skates or rays are used as a raw material, Asta’s process will yield “cartilage” or chondroitin. Chondroitin is commonly used as an over the counter nutraceutical for the treatment of arthritic joint pain.

Uses in the Marketplace

Asta’s PPP is applicable to the entire human food chain. It can be used in the $52 billion aquaculture feed market as an ingredient. It can be used as a ruminant animal feed ingredient and can be used by humans for direct consumption. The human market is very interesting since PPP can be a benefit in both the developed and developing worlds. PPP due to its high digestibility, high palatability, and low cost can be used as an ingredient to help reduce the 800 million chronically malnourished people in the world.  Also, it can be used as a protein ingredient in the $59 billion pet food/treat market. Omega-3 oils and calcium are expected to be used almost exclusively in the human and pet nutraceutical markets.

The two most common forms of concentrated protein ingredients for human consumption are soy and whey protein concentrates and isolates. These are sold almost exclusively in the human market because the manufacturing processes to concentrate and isolate soy and whey proteins are expensive. Thus, the sales price is too high for them to be used as feed. Conversely, the manufacturing cost of PPP is low in comparison. It is anticipated that the sales price of PPP will be such that it can be sold at a competitive price as an ingredient for aquaculture and livestock feed. Likewise, PPP is forecasted to have considerable applications in the human food and beverage markets as an ingredient to conveniently and cost effectively increase protein content. Additionally, soy protein is a known cause of allergens in humans and pets. Since PPP has a long shelf life and is a powder, it can be shipped economically over long distances without refrigeration or specialized packaging. Research has demonstrated that polypeptides in the Dalton size range of PPP are hypoallergenic.

Patented and Proprietary Process

Asta’s proprietary knowledge incorporates the manufacturing process, which features unique enzymatic and drying methods. Asta’s technology is rooted in enzymatic hydrolysis, a process that has been known for decades. Enzymatic hydrolysis is defined as “a catalytic decomposition of a chemical compound by reaction with water, such as the breakdown of proteins into amino acids by the addition of proteases.” Asta’s process can be described as a mechanical stomach that replicates the process of digestion, i.e. it breaks apart the polypeptide chains.  Asta’s food biotechnology is a continuous flow process that takes approximately one hour to turn its raw material into valuable products that fit into the fabric of life. Asta’s advanced protein product, PPP, is manufactured to a specific chain length range, and as such is engineered for maximum digestibility for specific animals, marine species, companion pets, and/or humans.

Of particular significance is that Asta has unlocked the method of drying the liquid hydrolysate (the liquid form of PPP) in a very cost effective methodology. This breakthrough enables Asta to produce its products at a cost point low enough to be commercially attractive, thus breaking the barrier, which up until now prevented hydrolysates from being widely accepted. Because the processor is automated, manpower requirements are minimized and nominal. Current efficiency level of the processor is 31% yield of sellable products. The remaining 69%, returns to the environment as clean pasteurized water, resulting in few, if any, environmental issues. Asta’s proprietary and patented process can reduce discarded food residuals that are a source environmental pollution.

Asta has developed a prototype 600lb/hr-input processor that has produced 93,000 pounds of PPP. After funding, the prototype processor will include design changes to increase efficiencies and to develop new processes. Asta intends to locate its much larger commercial processors near sources of raw materials.

Because the processor is automated, manpower requirements are minimal, and that the cost of raw materials are very low, it is forecasted that Asta’s manufacturing cost will be correspondingly low. Asta’s process is notably advanced and efficient. Asta’s process is versatile and is capable of using other raw material sources and different enzymes.

Conclusion

The science behind Asta’s products is significant.  Extensive scientific studies performed on peptides, di-peptides, and tri-peptides have validated that Asta’s PPP is a highly desirable form of protein, is digestible, and is hypoallergenic. Asta’s short chain “protein” powder is a foundational nutritive-ingredient for life; from microbes to mankind and everything in between.

Asta’s proprietary, continuous enzymatic hydrolysis process transforms raw materials into polypeptides with superior bioavailability. The company creates several products for many market segments.

Asta’s PPP is suitable for use in the entire human food chain; direct human consumption, aquaculture feed, and animal feed. Additional applications include the pet food and treats market as well as use in organic fertilizer. Because all of its markets are immense and global in scope the yearly economic collective value is virtually incalculable.

Asta has a unique drying methodology which, for the first time, makes it commercially feasible to produce a powder from liquid hydrolysate. As such Asta also has the opportunity to, in the long term, drive innovation and standards in this new field. Asta’s revenue and profit potential is enormous.

[1] Protein Requirements, Fish Nutrition. John E Halver, Ronald W Hardy. Academic Press, page 145. — 2002

[2] BP Nutrition — Aquaculture Research Centre: Animal and Fish Feeding Trials. — 1991, 1992

[3] Moore-Clark Diet Trial — Pink Salmon/R.W. Hardy, PhD, Affil. Associate Professor, School of Fisheries, University of Washington, Seattle, Washington. — 1991

[4] Cahu/Zambonino — Protein Hydrolysate Studies on Fish Larvae — 1994, 1997, 2001, 2003

[5] Larval Fish Nutrition. G. Joan Holt, page 396. — 2011.

[6] Hydrolyzed Proteins and Hypoallergenicity — Boye & Godefroy, 2010; Veternary Healthcare Symposium. — 2011.

[7] Fish meals, fish components, and fish protein hydrolysate as potential ingredients in pet foods. Folador, J.F.; Karr-Lilienthal, L.K., et. al., Journal of Animal Science, 84:10, pp 2752-2765. — 2006.

[8] Allergen Management in the Food Industry. Joyce Boye, Samuel Godefroy.  pp 218-219. — 2010.

[9] Protein Hydrolyzates From Aquatic Species, Fereidoon Shahidi, Xiao-qing Han, et al, Seafood Science and Technology Society, 18^th Annual Conference. — 1993.

[10] Hydrolyzed Proteins and Reparative Properties. — 2005

[11] Cahu/Zambonino. loc. cit.

[12] Cahu/Zambonino. op. cit.

[13] BP Nutrition — Aquaculture Research Centre. loc. cit.

[14] BP Nutrition — Aquaculture Research Centre. op. cit.

[15] BP Nutrition — Aquaculture Research Centre. op. cit.

[16] Fish meals, fish components, and fish protein hydrolysate as potential ingredients in pet foods. loc. cit.

[17] Hydrolyzed plant and animal proteins, Poultry Meat Processing, Edited by A.R. Sams, PhD. page 207. — 2001.

[18] Hydrolyzed Proteins and Hypoallergenicity. loc. cit.

[19] Allergen Management in the Food Industry. loc. cit.

[20] Protein Hydrolyzates From Aquatic Species. loc. cit.

[21] Hydrolyzed Proteins and Reparative Properties. loc. cit.

[22] Hypoallergenic Infant Formula. Grace, Victoria. About.com. tinyurl.com/7gg4dkk  Updated — 27 August 2008.

[23] Hypoallergenic Infant Formulas: Committee on Nutrition. American Academy of Pediatrics. Pediatrics, 106(2): pp 346-349. — Aug 2000

[24] Safety of a new extensively hydrolysed formula in children with cow’s milk protein allergy: a double blind crossover study. Terheggen-Lagro, Suzanne, Khouw, Ilse, et. al. BMCPediatrics, 2:10. — October 2002.

[25] “The Best Sources of Calcium in the Five Food Groups.” Oregon Dairy Council, Nutrition Education Services.  — 1994.