Working with Enzymes in Confections Enzymes play an essential role in making confections. Optimal conditions for their use are critical to ensure quality end product.

David Jordan Centerchem, Inc.

nzymes are nature’s catalysts for initi - acids. On this large molecule, a specific Eating biochemical reactions. They are region exists where substrate molecules bind present in all living organisms regulating var - and undergo various chemical reactions. ious chemical reactions and all aspects of cell This region is known as the active site (Fig - metabolism, including the digestion of food. ure 1). Essentially, the active site consists Enzymes allow larger nutrient molecules to of a section on the enzyme that form tem - be broken down into smaller molecules to porary bonds with a compound and in doing be metabolized. More than 5,000 reactions so, catalyze a reaction. catalyzed by enzymes have been identified. A particular enzyme can only do its unique David Jordan is the The primary enzyme used in confec - work with a specific compound and only director, technical sales tionery production is invertase, which con - under the right conditions. A lock and key and services for Cen - terchem, Inc. His respon - verts sucrose into glucose and fructose. It analogy is often used to describe the speci - sibilities include ingredi - is necessary to soften or control crystalliza - ficity of how any given enzyme, the lock, ent sales and technical tion in product centers, and for other uses will react on a compound, the key. This reac - support. He has been as well. Herein is an overview of enzymes tion process is illustrated in Figure 2. In this with the company for 16 years and previously including their chemistry and processing analogy, only a correctly sized substrate key held positions at Her - will fit into the active site keyhole on the conditions, and how to use invertase to gen - shey Foods, Colorcon erate desired results in confections. Other enzyme to unlock it. Once done, the subse - and Enzyme Develop - enzymes used in confectionery applications, quent reaction steps can occur. In reality, ment Corp. and the methodology and benefit of their the lock and key are based upon specific uses, are outlined as well. chemical bonds and structures. The nomenclature for enzymes is typi - AN OVERVIEW OF ENZYMES cally the compound that is being catalyzed, Enzymes are very large, three-dimensional also known as the substrate, followed by proteins, made up of several hundred amino suffix ase , derived from a Greek word mean - ➤

The Manufacturing Confectioner • January 2019 47 Working with Enzymes in Confections

Within a given ing to separate . Examples of these include fering reactions are the essence behind the category, there can be amylases (amylose), proteases (proteins), vast array of cheeses we enjoy. different enzymes lipases (lipids or fats) and cellulases (cel - In similar fashion, during the cocoa bean that will react with lulose). There are exceptions to this cate - fermentation process, naturally occurring the specific variations gorization. An enzyme can also have a microorganisms release their unique set of of the substrate. more specific function, such as lactase, enzymes that generate various flavor com - which breaks down lactose into galactose pounds of chocolate. For this discussion, and glucose; and saccharase, which cat - understanding that enzymes work in a very alyzes the hydrolysis of sucrose into glu - specific manner is what is most important. cose and fructose. The enzyme saccharase In these examples, the enzymes are clas - is more commonly known as invertase in sified as hydrolases because hydrolysis the confections industry. occurs, resulting in a splitting of the com - Within a given category, there can be pound. However, not all reactions involve different enzymes that will react with the splitting. Examples of other enzyme reactions specific variations of the substrate. For can be ligases, which combine molecules; instance -amylase randomly hydrolyzes transferases, which transfer specific functional α the -1-4 bonds on amylose, while glu - groups from one molecule to another; and α coamylase hydrolyzes -1-4 and -1-6 isomerases, which rearrange atoms. α α bonds stepwise, starting from the non- In the human body, our saliva contains reducing end. Similarly, different lipases amylases, which help break down starches will react and cleave different fatty acids, as soon as they enter our mouth. Our yielding unique flavor profiles. These dif - digestive tract, including liver, pancreas and intestinal flora, all generate various Enzyme Molecular Structure enzymes to aid in digestion. The lack of Amino Acid Chain these enzymes can manifest in various mal - adies, primarily digestion issues, such as gas and bloating. Active Site There are many dietary supplements based on the concept of ensuring there are enough digestive enzymes in our systems. There are also specific products such as lac - tase for those who are lactose intolerant. A common supplement, alpha-galacdosidase Figure 1 aids in the digestion of certain typically indi - gestible compounds (commonly found in Lock and Key Model of Enzyme Reaction beans), which will ultimately sit in the gut and “ferment” with the unpleasant side effects we are undoubtedly familiar with.

ENZYME SOURCES AND PROPERTIES Commercially, enzymes are derived from three primary sources: plant, animal and microbial (Figure 3). Those derived from Figure 2 plants and animals have specific properties ➤

48 January 2019 • The Manufacturing Confectioner Working with Enzymes in Confections while those from bacteria have a much do not need to be identified on a finished Another unique broader range of applications. Enzymes product label if they are deactivated. If property of enzymes also have the ability to undergo various however, the product does not go through is that they are not genetic modifications to help generate spe - this denature step, the enzyme used in the consumed in a cific functionality and properties. When process should be indicated. chemical reaction. enzymes are manufactured or purified, There are many types of enzymes used they typically go through a standardization in the food industry for the production of step where they are diluted for a specific a wide variety of products. Yeast is basically activity. The activity is a measurement of a bundle of enzymes used in making bread, the amount of active sites present, usually beer, wine and other fermented products. stated per gram. Yeast has many enzymes that break down Some microorganisms are used to pro - the different starches and sugars making duce a wide range of enzymes. For instance, them more easily fermentable. Fruit juice the enzymes pectinase, cellulase, amylase, production utilizes cellulases and pectinases lipase and even invertase can all be com - to optimize yield and clarify the juice. And of course, there is invertase, the primary mercially derived from the organism Sources of Enzymes enzyme and key ingredient used in the pro - Aspergillus niger . Based on the production 1. Fruit and Plant Sources method used, there will be a main activity duction of many confections. • Papaya –Papain • Pineapple –Bromelain and then, lesser side activities. However, a USING INVERTASE IN CONFECTIONS • Figs –Ficin selected strain of the yeast Saccharomyces 2. Animal Sources Invertase is an enzyme that converts the cerevisiae will produce an invertase with • Egg white –Lysozyme disaccharide sucrose molecule into its • Calf stomach –Rennet no side activity. monosaccharide components of glucose 3. Microbial Sources Another unique property of enzymes is • Yeast (Saccharomyces) and fructose (Figure 5) and as mentioned that they are not consumed in a chemical • Fungi (Aspergillus) earlier, this conversion plays a critical role • Bacteria (Bacillus) reaction. The enzyme finds a substrate to Figure 3 in confection making. This process is called act upon and then moves on to the next substrate. The enzyme remains active until Temperature and pH Impact on Invertase Activity it runs out of substrate sources. Because of this property, the amount of enzyme added is directly proportional to the time it takes to complete the reaction. If you have an enzyme with an activity of 10,000, it will take one-tenth the amount of time to work as the same enzyme with an activity of 1,000. The first chart shows solution at the optimal pH of 4.5; observe what happens when Every enzyme also has a unique set of temperature changes. Second chart shows solution at the optimal temperature; ob - serve what happens when pH changes. temperature and pH ranges at which they Figure 4 are optimally effective. Outside of these ranges, the enzyme effectiveness decreases, ultimately becoming completely ineffec - Invertase Hydrolyzes Sucrose into Glucose and Fructose tive. If enzymes are heated beyond a cer - tain point, they are denatured and irre - versibly deactivated as indicated in the Figure 4 graphs for invertase. When used in food processing, enzymes Figure 5 ➤

The Manufacturing Confectioner • January 2019 49 Working with Enzymes in Confections

During warehousing inversion. Sucrose has limited solubility in mon application is to soften a product that or en route to water at room temperature. Based on the needs to start out firm. Examples of this retailers, the concentration, it will crystalize in a short are a chocolate truffle or a chocolate-cov - invertase in the timeframe. Because glucose and fructose ered peppermint patty that is either product will begin to have a greater solubility in water, they will panned or enrobed. A hardened center is invert the sugar. no longer crystallize after the sugar has made to withstand the panning process or been inverted. transport through an enrobing line. The Commercially available invert syrup is center needs to maintain its structural a high-solid content liquid that has gone integrity to enable it to be processed, pack - through this inversion process and is used aged and shipped. During warehousing or in food and beverage applications where en route to retailers, the invertase in the crystallization is undesirable. The tradi - product will begin to invert the sugar. As tional method of producing invert syrup the sucrose is inverted, becoming glucose is to add acid to the concentrated solution. and fructose, the solubility increases and Upon completion of the inversion process, the centers begin to soften. The finished an alkali is added to raise the pH. This pieces are kept at a temperature that is process generates invert syrup that is eco - well below optimum and therefore, it takes nomical, but tends to have off colors and much longer for the inversion to complete flavors and cannot be considered organic. compared to invert syrup. This total time Enzymatically-produced invert syrup can be easily adjusted by modifying the yields a much cleaner and a potentially dosage of invertase used. organic, product. Holding the syrup at an The second way invertase is used in con - elevated temperature, with a high enough fections is to control crystallization. An dosage of invertase so that the inversion example is a cordial cherry, a cherry in a is complete in anywhere from 8 to 24 liquid surrounded by chocolate. Pieces are hours, produces a more purified end prod - often panned using a sugar solution that uct. Manufacturers must weigh the cost of contains invertase, but allowed to first crys - processing time and the energy used versus talize so they can be chocolate panned. cost of adding more invertase. As stated Other cordial cherries made via depositing previously, twice the invertase will enable are still in a viscous sugar fondant mixture the reaction to take place in half the time. enabling the pieces to complete the enrob - Invertase is used in confections in two ing process. In both cases, the invertase primary ways (Figure 6). The most com - will liquefy the saturated sugar, allowing

Invertase Use in Confectionery

Softening center after panning Controlling crystallization for panning or enrobing and extending shelf life Figure 6 ➤

50 January 2019 • The Manufacturing Confectioner Working with Enzymes in Confections the finished cordial cherry to be sur - points on issues that can arise, are outlined Invertase can be rounded by its requisite liquid syrup. in Figure 7. As an example, the center of a derived from both a Invertase can also prevent the product chocolate truffle may require cooking or fungal and a yeast from crystallizing over time, thus extending heating. If the invertase is added too soon, source. These two shelf life. An example is a chocolate truffle it could easily become deactivated resulting sources have slightly made via a one-shot depositor. The ganache in centers that remain hard. The invertase different pH and center is fluid enough so it can be easily must be added as the preparation cools temperature ranges deposited, however, this type of product well below the deactivation temperature. and, in turn, has a tendency to begin crystallizing as it The order of invertase addition can also advantages to one sits. This causes the thin, fluid center to be a factor. If an acid is being used, it is versus the other. become harder and grittier, and thereby important to add the invertase after the less desirable. The invertase will prevent, acid has been thoroughly mixed. If not, the or at least significantly slow, this process. invertase will come in contact with a highly Invertase Use Considerations acidified area and thus become denatured. In the above applications, and in most con - Invertase can be derived from both a fections where invertase is used, the product fungal and a yeast source. These two is never heated past the deactivation tem - sources have slightly different pH and tem - perature of the invertase, which is typically perature ranges and, in turn, have different 190 °F. Because the invertase is not deacti - advantages. For instance, the optimal pH vated, it must be identified on the final pack - of yeast-sourced invertase is 4.5 compared age labeling. to 3.0 for fungal-sourced. If a lower pH As previously mentioned, enzymes have product is being produced, the fungal- a specific temperature and pH range where sourced invertase type may be preferred. they function optimally. Beyond that, the Caution must be used with invertase from enzyme will be either less effective or a fungal source due to the possible pres - totally deactivated. ence of lipase, which could generate unde - These are important considerations when sirable side effects. If the confection con - determining the initial preparation and tains fat, lipase could cause the confection manufacturing of a confection containing to take on a soapy or cheese-like flavor invertase. Additional processing consider - and an off odor. ations when using invertase and enzymes This side effect can also occur if inclu - in general, along with troubleshooting sions or spices are used in the confections Considerations and Troubleshooting that have not been properly processed.

Processing Considerations Lipase can begin hydrolyzing the fat in the • Order of addition piece and cause a soapy off-taste. This has • Cooking temperatures • Shelf life and transit time been noted in some starch-moulded gum - • pH of center • Labeling mies and fruit snacks. If the starch is not • Unwanted enzymes properly dried, residual moisture and bac - Troubleshooting • Other ingredients processed with enzymes teria in the starch will remain on the piece. • Source of enzyme After the piece is oiled, the bacteria can • Inclusions or spices • Improper starch cleaning generate lipase, which will cause the oil to Figure 7 yield an off taste. ➤

The Manufacturing Confectioner • January 2019 51 Working with Enzymes in Confections

The potential exists to OTHER ENZYME CONSIDERATIONS the ability to reclaim scrap is increasingly use enzymes in other It is critical that there are no residual important. Scrap can often be reused as aspects of enzymes in any other ingredient in the con - feedstock for subsequent batches, however, confectionery fection. Amylase enzymes are used to process only in limited amounts. It is possible to processing, however, starches and syrups. If any residual amylase use enzymes to break down sugars, these areas need to is present, it will cause hydrolysis of the starch starches, pectins or proteins (gelatin) that be further explored. in the confection. Similarly, if fruit juice is are present in confectionery scrap prior to used in a pectin-based candy, any residual being used as feedstock. pectinase from the juice processing can inter - Other compounds considered waste fere with the confection properly setting up. byproduct can be helped via enzymes. For These cases are rare, but should be consid - example, the fruit juice industry treats filter ered if there are unexplained reactions taking membranes with enzymes to help clean place within the piece. them, reducing the expense and manpower There are a few lesser-used applica - associated with frequent filter changes. tions for enzymes in the confectionery Such operations should be evaluated on industry. Manufacturers of chocolate whether the addition of enzymes is prac - sauce have found that when the product tical in increased time and cost compared sits in the container it can thicken, causing to current methods of handling waste. pumping to be harder, and the product However, with a basic understanding of to become less desirable. This effect is the functionality of these compounds, find - the result of naturally occurring starch, ing other beneficial uses for enzymes is pectin and cellulose — primarily from the likely very possible and would make inter - cocoa pulp and shell fragments finding esting studies. their way into the chocolate during the CONCLUSION early stages of the chocolate making In summary, enzymes are a natural way to process. Thalia Hohenthal and Michelle hydrolyze various organic compounds. Weinberg from Guittard Chocolate have Invertase, by way of modifying the chem - a patent on the use of an “enzyme cock - istry and solubility of sucrose, is prevalent tail” to help break down these com - in many confections. The benefits to using pounds for improved flow capabilities. Hohenthal, Thalia et al; June 30, 1998. High invertase as well as other enzymes are Cocoa/Chocolate Liquor Syrups with quite significant, as long as necessary pre - Improved Flow Properties; US patent cautions are taken and appropriate con - 5,773,056. ditions are employed to optimize their use. Using confections as a carrier for the The potential exists to use enzymes in enzymes as dietary supplements has other aspects of confectionery processing, proven challenging. There is an added cost however, these areas need to be further to this step which to date has not been war - explored. Enzyme suppliers can provide ranted, however technology is available to all the information needed to optimize the ensure proper enzyme encapsulation. use of these important and essential con - Another area of potential use for fectionery ingredients. n enzymes within the confectionery industry is to help process scrap. As companies strive to decrease environmental footprints, Presented at the AACT National Technical Conference

52 January 2019 • The Manufacturing Confectioner