Although enzyme immobilization began over 100 years ago, it remains one of the most essential industrial technologies today. This is because immobilized enzymes are highly versatile and have a wide variety of potential applications. Also, the enzyme immobilization process has been slowly perfected over the years, making it much simpler, quicker, and efficient than ever before. This has served as another motivating factor to utilize this process.

These days, EI has become intricately involved in industrial processes that are based on biotransformations. It has also become a vital element to cutting-edge technologies, such as genetic engineering and much more. With that in mind, let's take an in-depth look at enzyme immobilization and why it's important.

What are immobilized enzymes?

Before delving into the importance of enzyme immobilization, let's take the time to understand exactly what immobilized enzymes are. Simply put, an immobilized enzyme is an enzyme that is attached to a material that is insoluble and inert. Immobilizing an enzyme allows for an increased resistance to variables such as temperature or pH. It also allows the enzymes to be stationary throughout the process, which makes it much easier for them to be separated and reused.

Nevertheless, the qualities of the enzymes produced, largely depend upon the carrier type and the process used to immobilize them. The most common processes are adsorption, entrapment/encapsulation, cross-linking, or covalent bonding. The breakdown of these methods are as follows:

Adsorption

Adsorption is one of the simplest and quickest methods of reversible enzyme immobilization. This method involves a variety of non-covalent interactions. Once enzymatic activity grows stagnant, these enzymes can be gently removed from the support, so the support can be regenerated and attached to a new enzyme. This can help eliminate costs associated with the use of support materials.

The process can be completed by immersing the carrier in an enzyme solution, so it can be adsorbed physically. Alternatively, the protein may be dried on an electrode surface. Either way, this method is often preferable because it is inexpensive to complete and does not require the use of an activation reagent.

Moreover, the other main benefit of this method is that it promotes easy contact between the enzyme and the support. Also, there are typically minimal conformational changes made to the enzyme because the immobilization occurs in just a few minutes. This leads to high immobilized molecule activity retention.

While there are many advantages to using this method, there are also disadvantages to consider. For instance, the method is known for promoting weak links between the carrier and enzyme, which causes substrate contamination. Also, certain support materials can lead to enzyme denaturation.

Entrapment

On the other hand, the entrapment method involves covering or caging the enzymes in materials such as polymers, polymer/sol-gel composites, sol-gels, and other inorganic materials. Much like the adsorption method, this method is favored because it is inexpensive. It also allows for increased mechanical stability and makes for easy contact between the substrate and enzyme. However, unlike the adsorption method, this process is irreversible.

There are disadvantages to using this method, as well. Namely, the small matrix pores size reduces the mass transfer of substrate to enzymes. Another disadvantage is the fact that big pores allow for carrier leakage. Moreover, this process also promotes low loading capacities and material abrasion. However, this can be solved by cross-linking.

Cross-linking

Cross-liking is a lesser mentioned, irreversible method of enzyme immobilization. Also known as copolymerization, this process does not require the use of support as a means of preventing the loss of enzymes into the substrate solution. Also known as carrier-free immobilization, in this method, the enzyme essentially acts as its own carrier. This helps to eliminate all of the advantages and disadvantages associated with the use of various carriers. Most notably, it eliminates the likelihood that the carrier will cause activity to become diluted. This method is simple and inexpensive and is widely used in industrial and commercial preparation applications.

However, the disadvantages lie in the fact that the enzymes being used are often not pure. Also, the reagents used are known for modifying the enzyme, which can lead to a loss in general catalytic properties.

Encapsulation

The encapsulation method involves enclosing the enzyme in a membrane capsule, which is made up of semi-permeable materials, such as nylon or cellulose. The overall effectiveness of this method is based on the stability of the enzyme being used. This method is advantageous because it is inexpensive and allows for large amounts of enzymes to be immobilized.

However, the disadvantages of this method are that it that there is a limitation in terms of pore sizes and that only small substrate molecules are able to cross through the membrane.

Covalent bonding

Covalent bonding is one of the most widely used methods of enzyme immobilization. This irreversible process involves covalently binding proteins to insoluble support materials. Formed between enzyme amino acid residues and functional groups of carrier surface, the covalent bonds formed using this method are known for being stable intra/inter-subunit bonds. This method makes it possible for the enzymes to be reused in more cycles than the other immobilization methods. This method also assists in the prevention of enzyme leakage in reaction environments. The results of this process are exceedingly pure and have an increased level of thermal stability.

While this is, by far, the most favored method of enzyme immobilization, it also has a very notable disadvantage. In particular, this method of covalent bonding causes a decrease in enzyme movement and can also contribute to conformational changes. This can ultimately result in a loss of enzyme activity.

Benefits of using immobilized enzymes

Now that you better understand what enzyme immobilization is, as well as the processes used, let's take a look at some of the benefits of using immobilized enzymes. They are:

  • Reuse and recovery: One of the major benefits of enzyme immobilization is that it makes reusing and recovering enzymes a much simpler process.
  • Cost-effectiveness: As already mentioned, using immobilized enzymes can be very cost-effective. This is because the ability to reuse enzymes saves money on things such as production, labor, space, and more.
  • Versatility: Immobilized enzymes have a wide variety of potential applications.
  • Consistent supply of products to the market: Since having the ability to isolate and reuse enzymes makes it possible to create products of all kinds, immobilized enzymes play a major role in various markets' abilities to keep certain products on the shelves and available to businesses and consumers.

 

Other notable benefits of immobilized enzymes are:

  • High enzyme-substrate ratio
  • Process control improvement
  • An increase in functional efficiency of the enzyme
  • Less labor input
  • Minimized reaction time
  • Reduced opportunities for contamination in products created

Applications of immobilized enzymes

Now that you know more about what immobilized enzymes are and how they can be beneficial, let's take a look at the various applications:

Food Industry

One of the top applications for immobilized enzymes is in the food industry. Immobilized enzymes can be used to make many processes cheaper and easier to complete. Having the ability to reuse these enzymes can allow many companies and restaurant franchises to offer various food products at a discounted price. This is especially beneficial for those who are in fast food and other areas which are known for providing large amounts of lost-cost food on a regular basis.

In other words, immobilized enzymes play a major role in food processing and analysis. For instance, food additives such as high fructose corn syrup are an example of products that are created using immobilized enzymes. Also, immobilized enzymes are intricately involved in processed food staples such as starches and cheeses. They are also important for food preservation.

Brewing/Beverage Industry

Another major application for immobilized enzymes is in the brewing industry. For instance, the use of immobilized yeast cells made it possible for the industry to transition from making traditional batches to brewing continual batches. This has enabled the industry to save more money on production while also allowing it to make more money because the products are now much more readily available.

Additionally, this system has also been used to enable breweries to create higher-quality products. In particular, the brewing industry has used immobilized enzymes to create high-quality beer. Using these enzymes has made it possible for manufacturers to focus on color, flavor, and headspace pertaining to stout beer. Over time, they have been able to create products that consistently taste and look great.

Biomedical Applications

On the other hand, immobilized enzymes can also be used in the biomedical industry. The use of immobilized enzymes within the biomedical community has been on the rise since the 1950s. However, in recent years, there have been some advances made. Namely, manufacturers have figured out how to isolate and purify enzymes for several production cycles. It can allow for the reuse of lysozyme up to 100 times. This is useful because the high antibacterial activity can be used against the four types of pathogenic bacteria.

In keeping with that, there are also various uses for immobilized enzymes to help with things such as the treatment and diagnosis of a variety of illnesses and diseases. For instance, they can be used in the diagnosis and treatment of conditions such as Cancer, Leukemia, Acatalasemia, and Hyperuricemia. They can also be used to help create artificial lungs, artificial kidneys, artificial pancreases, and more.

Pharmaceuticals

Immobilized enzymes have also become intricately involved in the pharmaceutical industry. Enzymes such as streptokinase, urokinase, asparaginase, deoxyribonuclease I, hyaluronidase, pegademase, glucocerebrosidase, and cysteine proteinases are all very important to the pharmaceutical industry. Therefore, having the ability to immobilize enzymes has proven to be vital to the past, present, and future of the pharmaceutical industry. For example, immobilized enzymes have been used to create life-saving medications such as penicillin. This has made the antibiotic more readily available across the globe.

Textile Industry

Also, immobilized enzymes can be used in the textile industry. This is one of the fastest-growing markets in terms of EI. This is largely due to the fact that these enzymes are known for their non-toxic and ecological properties. One of the primarily used enzymes in the textile industry is cellulase, which is used for cotton softening, denim finishing, laccase (for bleaching), and much more. Also, lipase is a commonly used enzyme that is commercially available for the textile industry. It can be used to create the conditions for better washing of cotton cloth. Either way, the main benefit of using these in the textile industry is that they are highly cost-effective, and they help make products cheaper and more accessible.

Wastewater Treatment

Additionally, immobilized enzymes can be used for wastewater treatment. Particularly, bacterial strains were immobilized to enhance the biological oxidation of toxic pollutants in wastewater. Also, horseradish peroxidase has been immobilized to remove phenol from wastewater at high levels.

Paper and Pulp Industry

The pulp and paper industry has also benefited greatly from using immobilized enzymes. However, this industry also faces some challenges in terms of finding ways to eliminate brown-colored specs and imperfections from the products. Nevertheless, as with many other industries, the use of immobilized enzymes makes for cheaper products that are more consistently available without unnecessarily wasting natural resources.

Detergent Industry

Lastly, the use of immobilized enzymes has been gaining traction in the detergent industry in recent years. For instance, alkaline protease is an enzyme that is commonly used in detergent. This allows the detergents to be more stable in higher temperatures, which makes for more efficient washing conditions.

Green Industrial Practices

"Green industry" is a term that applies to a wide variety of industries and sectors. Immobilized enzymes are an essential piece of global efforts to move toward more environmentally friendly and sustainable processes. In particular, the fact that they can be reused repeatedly and help reduce waste is one of the main selling points for using immobilized enzymes in this industry. For instance, manufacturers of bulk green-friendly chemicals are likely to use immobilized enzymes to make the product cheaper and more accessible while also reducing their carbon footprints.

Overall, enzyme immobilization is a process that has been around for over a century. Nevertheless, it continues to become increasingly relevant in a wide variety of industries. Although they are presently much more beneficial in certain industries than others, it seems that many industries understand the benefits of this process, and they are taking the time to develop them. Therefore, no matter what industry you are doing business in, you will probably need to become acclimated with enzyme immobilization sooner than later. Rather than waiting until you are forced to learn about it to stay competitive in your industry, taking the time to become familiar with enzyme immobilization in the present can make for a much more lucrative and efficient future.

 

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