Enzymes in food supplements - application and challenges

In recent years, there has been a growing interest in the use of enzymes as food supplements to improve digestive health and alleviate certain difficulties.

AUTHOR: Ana Gjergja Sekulić, M.pharm., univ.mag. phytotherapy and dietotherapy

Enzymes are products of various digestive system organs such as the pancreas, small intestine, and stomach. Their tasks in the body can be different - from breaking down complex molecules from food and facilitating digestion and absorption to reducing inflammatory reactions and managing various physiological functions.

Examples of enzymes that are most often found in food supplements

Digestive enzymes

Amylase helps breaking down carbohydrates into simple sugars that can then be easily absorbed by the body. It is usually used to help digest starchy foods.

• Protease helps breaking down proteins into smaller peptides and amino acids and helps with digesting protein­rich foods.

• Lipase helps breaking down fats into fatty acids and glycerol and is used to aid in the digestion of fatty foods.

• Cellulase helps breaking down the cellulose found in plant cell walls, making it easier for the body to digest plant­based foods.

• Lactase helps breaking down lactose, the sugar found in milk and dairy products, making it easier for lactose­intolerant people to digest this food.

Proteolytic enzymes

Bromelain is found in pineapple, and it helps breaking down proteins. It is often used as a supplement to reduce inflammation and swelling.

• Papain is found in papaya and helps breaking down proteins and, like bromelain, is used to reduce inflammation and swelling.

Systemic enzymes

They are the most numerous in the body and participate in all bodily processes such as breathing, speaking, moving, thinking, behaving, and maintaining the immune system. Systemic enzymes are commonly used to reduce inflammation, support cardiovascular health, and boost immune function. So far, they are less represented in the category of dietary supplements, but they certainly offer application potential considering the indications for which they are used. The most famous ones used as dietary supplements are:

Serapeptase

Nattokinase

Lumbrokinase

Production of enzymes

Enzyme preparations are usually produced by a microbial fermentation process. This involves growing specific strains of bacteria, yeast, or fungi in a controlled environment and then extracting the enzymes. The specific organisms used to produce enzymes differ depending on the type of enzyme being produced. For example, lactase enzymes are often produced by strains of the Aspergillus fungus, while papain enzymes are usually obtained from the papaya fruit.

There are several key challenges in the production process. This includes proper selection and the optimization of microorganisms that can produce the desired enzyme. It also includes identifying microorganisms that have the ability to produce large amounts of enzymes and optimizing their growth conditions. After microorganisms produce an enzyme, it must be purified to remove any unwanted components and to ensure that the final product is of high purity. This can be a demanding process, as enzymes are often produced in complex mixtures and can be difficult to separate from other proteins and cellular components. Enzymes can be sensitive to changes in temperature, pH, and other environmental factors. This is a very important item in the production of enzyme supplements because the final product must be stable and retain its activity over time.

Enzyme supplements can come in a variety of pharmaceutical forms, including tablets, capsules, powders, and liquids. Each of these forms presents a unique challenge in terms of formulation, as the enzyme must be protected from degradation and must be delivered in a form that is easily absorbed by the body. Enzymes produced by microbial fermentation can find their application in various branches of industry ­ pharmaceutical, cosmetic, food industry, and many others.1

Enzymes can be extracted from fruit and animal tissue. Many commonly used enzymes such as papain (from papaya fruit) and bromelain (from pineapple) are derived from fruit. While, for example, trypsin and chymotrypsin are obtained using the pancreas tissue of cows or pigs. Extraction of enzymes from fruit and animal tissue involves a process of tissue homogenization or maceration, followed by separation of the enzyme from the tissue by centrifugation or filtration. The enzyme extract thus obtained can then be purified and concentrated and used for various industrial or research applications. 2

Enzymes can also be produced by genetic engineering techniques. Thus, for example, proteases, amylases, lipases and cellulases3, and certain systemic enzymes can be produced.

A question of quality

The quality of enzymes can vary depending on the technology used for their production. Enzymes produced by modern fermentation technologies such as microbial fermentation or recombinant DNA technology are usually of higher purity than those produced by traditional methods such as extraction from animal or plant sources. Also, modern technologies enable better control over conditions such as pH and temperature, can increase stability and activity, and ensure better microbiological purity. This results in a more effective and efficient enzyme.

When evaluating the quality of an enzyme, it is important to consider these factors in relation to the specific application for which the enzyme will be used. For example, an enzyme used in the food industry must meet strict purity and safety standards, while an enzyme used in a research laboratory must have certain activity or specificity characteristics. Overall, the technology used to produce enzymes can have a significant impact on their quality, and choosing the right production method can help ensure that the enzyme is efficient, safe, and consistent.

Given that both the stability and activity of enzymes are very sensitive to changes in temperature, pH, ionic strength, proteolysis, oxidation, aggregation, and other factors, production technology, formulation, and storage are key to choosing a quality enzyme. To ensure a long shelf life, enzyme supplements are usually formulated with stabilizing agents such as buffers, salts, sugars, and surfactants that can protect the enzyme from degradation and maintain its activity over time. Likewise, techniques such as microencapsulation and lyophilization can contribute to an extended shelf life.4

Application of enzymes

Digestive enzymes

Supplementation with digestive enzymes is most often used to treat digestive problems such as flatulence, dyspepsia5, and abdominal pain because they reduce the load on the digestive system by the more efficient breakdown of food6. They are also used when the absorption of nutrients needs to be improved, for the same reason. Digestive enzymes can help people with food intolerances such as lactose intolerance7. They can also be used in various inflammatory bowel diseases, especially irritable bowel syndrome8. Also, indigestion that occurs in the state of pancreatic insufficiency can be alleviated by using digestive enzymes.

Proteolytic enzymes

Bromelain and papain are two proteolytic enzymes that are most often found in food supplements. The application of bromelain is diverse. It is most often used to reduce inflammation and swelling. Given that it has anti-inflammatory and analgesic effects in addition to anti­edematous, anti­thrombotic, and fibrinolytic9 effects, it is used for various conditions from osteoarthritis, to sinusitis and sore throat. The mechanism of action of bromelain is mediated by the following factors: increasing fibrinolytic activity in serum10, decreasing fibrinogen levels in plasma11 , and decreasing bradykinin levels (resulting in reduced vascular permeability) and therefore reducing edema and pain12 by mediating prostaglandin levels (reducing PGE2 and thromboxane A2 levels).

The body can absorb bromelain to a significant extent without negatively affecting its proteolytic ac­

Nutramedic &Cosmetics

tivity. Its many therapeutic benefits include wound debridement, improved drug absorption, and the treatment of sinusitis, bronchitis, angina pectoris, surgical trauma, and thrombophlebitis. In addition, it shows a beneficial effect on numerous cardiovascular conditions. Bromelain also promotes apoptotic cell death and exhibits some anticancer properties13

The use of papain is very similar to that of bromelain. Papain acts as a debridement without harmful effects on healthy tissues due to enzyme specificity, acting only on tissues lacking α1­antitrypsin plasmatic antiprotease that inhibits proteolysis in healthy tissues14. Papain has proven to be extremely useful in dentistry during the process of dentin excavation15 and chemomechanical caries removal16. The enzyme papain has a long history of use in the treatment of sports injuries and other causes of trauma, where studies have shown that minor injuries are healed faster with papain proteases than with placebo17. Papain is also successfully used to treat allergies associated with the leaky gut syndrome, hypochlorhydria, and intestinal disorders such as gluten intolerance. It has also been noted that it finds its place in the treatment of symptoms of acute allergic sinusitis and toothache18

Metabolic enzymes

Serratiopeptidase is a metalloprotease with a molecular weight of 45–60 kDa. It belongs to the Serralysin group. It is originally obtained from Serratia marcescens isolated from the intestine of the mulberry silkworm BombyxmoriL. The clinical use of the enzyme has been recorded for many diseases such as arthritis, sinusitis, inflammatory bowel disease, and bronchitis19. Other applications in clinical practice mainly include breast diseases, atherosclerosis, Alzheimer's disease, sinusitis, hepatitis, lung disorders, and uterine fibroids20. As the demand for said enzyme has increased, certain safety issues arose. Given that the biomass used for enzyme production is considered dangerous, techniques for recombinant enzyme production in E. coli21 have been developed.

Nattokinase is a purified serine protease extracted from natto, a traditional Japanese food produced by the fermentation of soybeans with the bacterium Bacillussubtilis(natto). In 1980, Hiroyuki Sumi, a Japanese researcher, discovered that natto could dissolve artificial fibrin. Further research revealed that it not only breaks down fibrin but also the substrate of plasmin22. Nattokinase can degrade blood clots by directly hydrolyzing fibrin and plasmin substrates, converting endogenous prourokinase to urokinase (uPA), degrading PAI­1 (plasminogen activator inhibitor­1), and increasing tissue plasminogen activator (t­PA) which supports fibrinolytic activity23. As a dietary supplement, nattokinase is used for blood thinning, preventing the formation of blood clots, and improving blood circulation24. Studies also show that nattokinase can alleviate other diseases such as hypertension25, stroke26, Alzheimer's disease27, and atherosclerosis28. The use of nattokinase is considered to be safe, and the production by fermentation of the bacterium B. subtilis does not present a problem in the form of biomass toxicity. However, as the demands of the market grow, new ways of producing this enzyme using recombinant technologies are also being researched.

Lumbrokinase is a group of enzymes isolated and purified from different types of earthworms. They are recognized as fibrinolytic agents that can be used to treat various conditions associated with thrombosis. Advances in genetic technology have enabled the production of recombinant lumbrokinase and made it feasible to purify a single lumbrokinase enzyme for potential antithrombotic use29. Lumbrokinase is very specific for fibrin as a substrate and does not cause excessive bleeding30,31. It can dissolve fibrin itself or convert plasminogen to plasmin, by inducing the endogenous activity of tPA to dissolve fibrin clots32,31. This is precisely why lumbrokinase is used in various conditions such as the risk of thromboembolism33, thrombosis therapy34, cardiovascular diseases, stroke35,36, and all other conditions that require circulation support. The application also shows potential for reducing intra­abdominal adhesions. It is a complication after abdominal surgery caused by suppression of fibrinolytic activity and increased invasion of fibroblasts into the injured area, which can lead to chronic diseases such as chronic pain, intestinal obstruction, and female infertility37.

Conclusion

The use of enzymes in food supplements offers a very wide range of indications. Many enzymes and their applications are already well known, while so­ me new ones, especially systemic enzymes, are rising stars. Research is going in the right direction, so the future of this type of food supplement is certainly promising.

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