Breaking DownEnzymes

Enzymes are a cornerstone in digestion, but as more studies are conducted on the many enzymatic compounds that exist in the food we eat, as well as animals, plants and fungi, researchers are discovering they play a much larger physiological role than just digestion.

The trouble is, enzymes are unstable in nature and under certain conditions. "Prolonged exposure to extreme heat and humidity can denature enzymes more rapidly and result in loss of activity," said Scott Ravech, CEO, Deerland Enzymes Inc. "Enzymes also lose activity due to oxidation and/or hydrolysis. These phenomena are inherent in the enzyme, and there is little that can be done."

"It is important to understand that enzymes are proteins with very complex structures," said Mike Smith, vice president, Specialty Enzymes and Biotechnologies. "These proteins have a very complex shape created by many different kinds of bonds. It only requires the slightest change to this shape, and an enzyme is no longer active."

Because of these inherent characteristics, enzymes need to be carefully handled, stored, formulated, encapsulated, etc. "It is recommended that enzymes be stored in a cool, dry environment. Other conditions such as the high acidity of the gut where some enzymes may be sensitive can be overcome with enteric coatings or microencapsulated enzymes," Ravech added.

Mark Anderson, Ph.D., director of research and development, Triarco Industries Inc., agreed, echoing: "When it comes to enzyme formulation, stability is key. Test for stability and activity, and then test again if excipients have been added. Even seemingly inert ingredients can create differences you need to allow for in your formula. And remember, while an enzymes sensitivity to its environment may mean taking extra care in storage and processing, it is that same highly responsive nature that makes an enzyme so finely attuned to the digestive processes it supportsif it is properly formulated. For instance, a digestive enzyme can be prematurely activated or destroyed in the acid of the stomach, rendering it useless for nutrient absorption."

Next: Enzymes in food and beverages

F&B

Due to stability and survival issues, enzymes have yet to really break into the food and beverage category, and are mainly delivered via supplements. "Meal replacement and high-protein shakes are the one area where enzymes are commonly used," Smith said. "The advantage here is these are typically sold in powder form rather than in a ready-to-drink format. The powder is mixed with water or juice at the time of use, so enzyme activity will be at its maximum." Smith noted the challenge with enzymes in ready-to-drink products and most foods is two-fold: "First, once an enzyme goes into solution, it is active; once it's active, it has a limited time before it denatures. Therefore, shelf life is rather short. The second issue is heat. Most foods and beverages are pasteurized, which also results in the denaturization of enzymes."

Ravech also discussed the issue of potency, stating: "In the case of functional food, where high heat may be used as part of the process, enzymes would have a difficult time maintaining their potency. For cold-oil or lower-temp processes, enzymes may be considered, but would also have to be evaluated for their impact on quality and stability of the finished goods. That said, a manufacturer should work with a company competent in enzyme activity assays to work side by side with them to ensure the integrity and potency (activity) are in fact maintained during and after manufacturing."

Ravech said enzymes' inclusion in beverages isn't a black-and-white situation. "It depends," he said. "In order for enzymes to function they are constantly bending and stretching, which, like anything else, results in the wearing out (denaturing) of the enzyme. The more freedom an enzyme has, the quicker it will denature. Enzyme stability depends on its environment, so having limited/restricted freedom (as in powder form) means enzymes usually maintain higher activity. Beverages that are always in a water-based solution will have a negative impact on the enzyme by denaturing it fairly quickly due to the freedom in which the enzyme 'stretches' in the solution."

Next: Enzymes benefit to digestion

Break Down

Enzymes are prolifically used to address digestive issues. In short, digestive enzymes hydrolyze nutrient substrates: amylase enzymes break down carbohydrate substrates, lipases handle fats and proteases take care of proteins; other types include cellulases, pectinases and xylanases. The proteases, also known as proteolytic enzymes, include the pancreatic enzymes chymotrypsin and trypsin, as well as the plant-derived bromelain from pineapple and papain from papaya.

One of the most well-known enzymes supplements is BEANO® (from GlaxoSmithKline)alpha-galactosidase used to control gas and bloating. Another enzyme that addresses one of BEANO's targeted food groupsbeans and legumesis Legumase® (from Triarco). "Legumase targets specific sugars in beans and other legumes that make these foods uncomfortable to digest," Anderson said. "This is because our bodies actually lack the enzyme needed to break down such sugars. It allows many consumers to eat the healthy foods they love and make dietary choices they feel good about, without fear of digestive upset or embarrassment."

Due to a rise in digestive issues such as indigestion due to lactose or gluten intolerance, or conditions such as irritable bowel syndrome (IBS), supplements such as lactase have become a popular remedy. "Digestive blends are now being marketed for dairy, gluten or for detox programs, among others. In some cases, they are being combined with probiotics as well," Smith said.

In populations where meat is a staple of the diet, help with digestion of meat proteins is desired. Papain and bromelain are both meat tenderizers, due to their ability to handle tough meat fibers, and they are also popular after-meal enzyme supplements.

Deerland Enzymes is conducting research on the use of enzymes, as well as chelating agents, peptides and essential oils that promote natural gastrointestinal (GI) flora health and well-being. "We are excited about the possibility that this research will give us greater insights into conditions like small intestinal bacterial overgrowth in patients with IBS," Ravech said. "In the past decade, clinical investigators have revealed more than 50 percent of the individuals with the functional diagnosis of IBS are afflicted by an overgrowth of bacteria in the proximal small intestine, thereby causing alterations in the structure and function of the absorptive small intestine. Deerland Enzymes, working in close conjunction with Theodore Hersh, M.D., AGAF, MACG, have conducted in-depth research into this condition that may suppress the growth of colonic microorganisms."

Even though lactose intolerance has been well-addressed and research is ongoing for supplements that target IBS, gluten intolerance is only beginning to pick up steam. In late 2010, scientists from the University of Salermo and the European Laboratory for the Investigation of Food-Induced Diseases at the University of Naples Frederico II, Italy, published a report in Enzymes Research on the various methods being explored to use enzymes in the detoxification of gluten and its protein constituent gliadin.1 Enzymes can break down the peptides from gluten that are resistant to the bodys endogenous proteases in people with gluten intolerance. However, these enzymes can be deactivated in the acidic stomach environment; encapsulating these enzymes does little good, as they can not efficiently break down gluten peptides before they reach the small intestines, where they cause the most trouble.

Enzymes are reaching beyond the belly and science is now studying their systemic mechanisms. "Enzymes have benefits that extend beyond their direct effects on digestion, and emerging research tells us they may be useful in a number of applications people may not typically associate them with," Anderson said.

"Systemic enzymes also tend to be positioned for condition specific use," Smith agreed. "These include anti-inflammatory issues, joint health and cardiovascular health. The systemic enzymes are an exciting area of investigation. Systemic enzymes are predominantly protease enzymes."

Next: Enzymes for immune health

In the immune system, enzymes are prized for their ability to break down proteinsmany would-be pathogens, such as bacteria and the outer shell of viruses, are protein-based. Proteases can break down such protein invaders to the body, but they can also help activate immune cells.

On the flip side of protein and proteases, "Loss of lean muscle mass can affect the immune system, the risk of osteoporosis and injuries such as hip fracture, and the ability to maintain independence later in life," Anderson added. "This loss of muscle can be the result of inadequate protein intake. Older adults may find it difficult to consume all of the protein they need because of difficulty chewing protein sources such as meat. And because we produce fewer endogenous enzymes as we age, they may not be absorbing all of the protein they do eat. Because Aminogen® helps to break down protein and increase amino acid absorption, it offers these consumers a new way to support a healthy protein intake." A clinical study published in the Journal of the International Society of Sports Nutrition showed Aminogen increased amino acid levels by 100 percent, branched-chain amino acid levels by 250 percent, and nitrogen retentionan indicator of protein absorptionby 32 percent.5

Protease also affects the flow and health of blood factors in cardiovascular health. "The use of enzyme-based formulations for anti-inflammatory and heart health are key areas of focus for many consumers and have been an area of significant R&D effort for Deerland," Ravech said.

Excess fibrin can promote inflammation and plaque formation in blood and lymph vessels, blood clots and hardened tissue around varicose veins. Many proteases are considered fibrinolytic, including the primary blood-born enzymes plasmin and thrombin.

Specialty Enzymes's nattokinase (as NattoSEB®) and serratiopeptidase (as Peptizyme SP®)which helps silkworms digest their cocoonsexhibit fibrinolytic as well as anti-inflammatory activity.10-14 "The potential for use in inflammatory and cardiovascular conditions is significant," Smith said. "Our proprietary blend, Exclzyme EN, has demonstrated significant anti-inflammatory activity as well as a significant reduction in C-reactive protein (CRP)."

Additionally, results from a clinical study showed Aminogen also reduced CRP, a marker of inflammation in the body, by 10 percent as well as a reduction in blood pressure.15

Research is also confirming enzymes' role in energy and endurance. "With the leadership of Dr. Hersh (Deerland Enzymes' chief science officer from a joint development alliance with Thione International), we have been able to create a formulationThioZyme AOthat can replenish and maintain the bodys antioxidant defenses to reduce high oxidative stress, such as digesting food in the GI tract to producing energy in every cell of your body," Revech said. "Additionally, ThioZyme AO recycles spent vitamins C and E."

Triarco's Carbogen® enzyme, designed to target complex carbohydrates, increased available energy and timeto exhaustion by 43 percent, thereby increasing endurance;16 and a collection of pre-clinical animal studies confirmed adding Carbogen to a meal replacement bar increased the rate of glucose absorption.17 Increased blood glucose levels were maintained over a period of five hours. "The finding could be significant for athletes and other consumers who look to carbohydrate products for a healthy boost, and for manufacturers who want to enhance their current energy or fiber formula," Anderson added.

Next: The future of enzymes

What's Next

While research is demonstrating enzymes' benefits go beyond the gut, this truth warrants more and continued research efforts. "I think the most important next step for enzymes is research," Anderson said. "While digestive enzymes have a long‐standing history of use and efficacy, research points to new systemic uses and benefits. As a scientist, I am eager to explore these further. I think we are just beginning to discover all of the ways enzymes can influence not only digestion, but metabolism, inflammation and cardiovascular health."

Custom and condition-specific formulas may be garnering more attention in the future. "It is our belief that customizable solutions and condition-specific supplements (in most cases, multi-component, enzyme-based formulations) will continue to evolve and are quickly becoming the norm," Ravech said. "Combining multiple productsenzymes with prebiotics, probiotics, botanicals/herbals, etc.enables our customers to offer consumers a broader spectrum of performance beyond what can be expected from basic or single component products. This trend demands companies embrace and are able to support customization to provide true value to their customers in helping to position new products across many facets of human health." Deerland created a no-cost service offeringYourBlenddesigned to provide formulators with a customization platform for specific performance benefit in dietary supplements.

Industry efforts and innovation will continue to spur this market, meeting consumers' digestive needs and more.

References listed on the next page.

Enzyme References:

1.       Caputo I et al. Enzymatic strategies to detoxify gluten: implications for celiac disease." Enzyme Res. 2010 Oct 7;2010:174354.

2.       Engwerda CR et al. Bromelain modulates T cell and B cell immune responses in vitro and in vivo." Cell Immunol. 2001 May 25;210(1):66-75.

3.       3. Engwerda CR et al. Bromelain activates murine macrophages and natural killer cells in vitro." Cell Immunol. 2001 May 25;210(1):5-10.

4.       4. Barth H et al. In vitro study on the immunological effect of bromelain and trypsin on mononuclear cells from humans." Eur J Med Res. 2005 Aug 17;10(8):325-31.

5.       Oben J, Kothari SC, Anderson ML. "An open label study to determine the effects of an oral proteolytic enzyme system on whey protein concentrate metabolism in healthy males." J Inter Society Sports Nutr. June 2008; 5:10.

6.       Taussig SJ et al. Bromelain: a proteolytic enzyme and its clinical application. A review." Hiroshima J Med Sci. 1975 Sep;24(2-3):185-93.

7.       10. Felton GE. Fibrinolytic and antithrombotic action of bromelain may eliminate thrombosis in heart patients." Med Hypotheses. 1980 Nov;6(11):1123-33.

8.       11. Metzig C et al. Bromelain proteases reduce human platelet aggregation in vitro, adhesion to bovine endothelial cells and thrombus formation in rat vessels in vivo." In Vivo. 1999 Jan-Feb;13(1):7-12.

9.       12. Fujita M et al. Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat." Biol Pharm Bull. 1995 Oct;18(10):1387-91.

10.   Fujita M et al. "Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan.". Biochem Biophy Res Com. 1993;197(3):1340-1347. 

11.   Pais, E. et al. "Effects of nattokinase, a pro-fibrinolytic enzyme, on red blood cell aggregation and whole blood viscosity." Clin Hemorheol Microcirc. 2006;35(1-2):139-42

12.   Fujita M, et al. "Transport of nattokinase across the rat intestinal tract.". Biolog Pharmaceu Bulletin. 1995;18(9):1194-1196.

13.   Moriya N,Nakata M, et al. "Intestinal absorption of serrapeptase in rats." Biotechnol Appl Biochem. 1994 Aug;20 ( Pt 1):101-8.

14.   Tachibana M,Mizukoshi O,Harada Y, et al. "A multi-centre, double-blind study of serrapeptase versus placebo in post-antrotomy buccal swelling." Pharmatherapeutica. 1984;8:526-30

15.   Ibid. Oben J, Kothari SC, Anderson ML. "An open label study to determine the effects of an oral proteolytic enzyme system on whey protein concentrate metabolism in healthy males." J Inter Society Sports Nutr. June 2008; 5:10.

16.   Frank L., Bear J., Lambert C., and Anderson M."The Effects of a Pre‐exercise Feeding With or Without Fungal Carbohydrases (Carbogen®) on Blood Parameters and Exercise Performance in Elite Cyclists: A Preliminary Study." Inter J Sports Nutr Exerc Metab. 2002;12:282‐290.

17.   Anderson, Mark L. "The Effect of a Dietary Carbohydrase Enzyme System on Blood Glucose Levels When Combined with Foods of Varying Glycemic Index in Male SpragueDawley Rats. J Med Food (14) 2011. [Epub ahead of print Aug 30]