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.