WHAT DISTILLERS SHOULD KNOW ABOUT ENZYMES

WHAT DISTILLERS SHOULD KNOW ABOUT

how ENZYMES WORK

Enzymes are powerful organic molecules necessary for all life’s functions in all living things. A good example of necessary enzymes are catalases, which are found in animals, ENZYMES including in the human liver. This enzyme performs its function so fast that without it, the chemical reaction would take more than 31 years to occur in its absence. Enzymes are unique in their source and Over the past decade and perhaps more, few products used in the production of potable spirits have been as divisive a choice as exogenous enzymes how specific they are to a certain function and can vary in how produced by biotechnology companies. Perhaps the growth of the craft spirits well they perform chemical processes industry and lack of training or other outside factors such as unknown under specified conditions, such as pH financial expenditures make the use of enzyme products an afterthought. and temperature. Enzymes are extremely Initial capital investments such as site build outs, equipment, fast in their process, however, the same enlicensing, and permits are every craft distiller’s number-one line zyme from two different sources can vary item(s). These are next followed by ingredient decisions greatly in how they perform. This, among — primarily sugar sources — for fermentation such as types of grains, sugar cane, and grapes. other factors, has a large impact on how an enzyme product performs better compared to a similar type product. The choice to use exogenous enzyme products is inherent to a company and craft distiller’s own goals, commonly associated with marketing the process and product. Not using enzyme products such EVOLUTION of ENZYMES in as a thermostable alpha amylase to hydro lyze starch results in lost yields that can af craft distilling fect a company’s bottom line. Depending

Without the distiller even knowing or un- on the conditions, desired outcomes or derstanding the basics of biochemistry or en- situation at hand, the use of enzymes can zymology, enzymes are a significant contribu- support craft spirit production as processtor to the process from beginning to end. First, ing aids to: malt is a cereal grain, most commonly barley, • Help lower energy input, that has been harvested, steeped, and kilned, • Reduce costs, allowing native enzymes to be expressed and • Save time and labor, and stored in the grain until further use in the mashing process. Yeast produce enzymes throughout • Use feedstock ingredients as their entire life cycle, enabling the microorgan- efficiently as possible to improve ism to bud, grow, and support life processes in yields and nutritional quality of their exposed environment. In addition, meta- fermentation, leaving very little on bolic processes in yeast for carbon and nitrogen the table as lost revenues.

uptake and emission are based on a series of enzymatic steps. How, when and where exogenous enzyme products are used allow a craft distiller to produce distillate in a way that is non-energy intensive, at a low cost, and in a timely manner. Thus, enzymes are already present in the process, but are they effectively utilized?

Enzymes have been studied for many years, largely driven by the brewing industry, to understand how and why processes were successful or unsuccessful to establish production consistencies. The etymology of “enzymes” derives from Latin “en” meaning “in” or “inside” and “zyme” meaning “yeast.” Zymology is the study of yeast. Enzymology is the study of what is inside yeast—enzymes! Major advances in enzyme products for craft distilling are built on decades of learnings and knowledgeable applications.

IMPLEMENTATION process

So, how does one approach the implementation of exogenous enzymes in a craft distilling process? This four-part series will walk through different groups of enzyme classes (First, thermostable alpha amylases, followed by glucoamylases, non-starch polysaccharide hydrolases and proteases) depending on application in the distilling process. Each enzyme class will have different environment requirements and function on different components of the feedstock.

Glucose is a six-carbon sugar with associating hydroxyl groups with the chemical formula C6H 12O6. In starch, glucose is linked as a polysaccharide (meaning “many sugars”) in either a straight chain orientation at the first and fourth carbon atoms and denoted as an alpha-1,4 bond (amylose) or in a branched chain at the first and sixth carbon atoms as an alpha-1,6 bond (amylopectin). The ratio of these bonds depends on the source of starch such as sweet corn versus waxy corn. When any cereal grains or root tubers are used for spirit production the first step in the process is milling to fracture the feedstock and access the starch granules. Next, a slurry mix with water and possibly a buffering component such as thin stillage/backset or acidulants such as lactic or citric acid is made, however this is not required. When starch is heated in the slurry mix, the starch crystals begin to swell allowing water molecules to enter. The starch swells and creates friction, leading to increased viscosity up to the point of gelatinization. Then, an enzyme called amylase is required to hydrolyze the linkages between each glucose molecule in the polysaccharide. An alpha amylase will hydrolyze, or break down, alpha-1,4 bonds at random locations while a beta amylase is more systematic in hydrolysis that results in maltose, a dimer (linked pair) of glucose. Native malt amylase enzymes are most active at 65 degrees Celsius and stable only a few degrees beyond this, around 70° to 75 degrees Celsius the enzymes are denatured by heat and all activity is lost. This improper handling of the malt in the mash cooking by heating beyond the temperature of the enzyme’s denaturation will result in the swelling of the starch without any viscosity break. To alleviate this, exogenous heat-thermostable enzyme products are employed to complete the hydrolysis of starch; these products are stable up to 85 to 95 degrees Celsius.

product CONSIDERATIONS

Returning the focus to what an enzyme is, these processing aids are an organic, naturally produced, specifically folded amino acid chain that possess a specific functionality. The performance of this specificity is optimized in a small range of conditions such as pH and temperature. Field biologists source organisms from all environments and conditions throughout the world. These include hydrothermal vents in the ocean, salt lakes, arid deserts, and tropical rain forests. Then, under laboratory and biofermentation conditions, enzyme potentials are exploited through a selective trial process.

Once “discovered,” enzymes are produced in expression systems, separated, clarified and filtered from any cell debris, then finally packaged for the consumer end use. Enzyme products from biotechnology companies are free from foreign DNA and contain no living microorganisms due to the clarification and filtration process monitored by rigorous quality assurance programs. A thermostable alpha amylase is a great example of an exploited enzyme sourced from a hyperthermophilic (meaning “very high heat loving”) bacteria produced for industrial process applications. Enzymes can take a substrate, for instance starch, and efficiently convert it to a product, a long-chain dextrin, in a continuous manner until all the substrate is depleted or until the environment becomes unfavorable for optimum activity, such as a change in pH or temperature.

The amount of an enzyme product used in a process is equally important. Too much could lead to excess cost and sometimes too much processing of substrate that could give unwanted results. If too little product is used, then the complete processing to the desired product may not be achieved or the time investment to completion may be too long. Trialing product for optimization under the guidance and recommendation of a technical assistant is important to achieve successful results.

For decades, enzyme products have benefited industrial processing. As industrial producers shift to more sustainable processes requiring lower inputs of energy and chemicals, enzymes provide a simple solution to an essential need. Just as they support life on this planet, naturally derived enzymes support the production of craft spirits through faster processing, using less energy input. Enzymes can help ensure that less product is wasted, thereby improving yields that lead to more product.

Dr. Nathaniel Kreel, Ph.D. received his doctorate in Biochemistry from Ohio State University and has worked inapplication technology and product development of enzyme products for the technical biofuel and potable spirit industries. He can be reached at Nathan.kreel@univarsolutions.com.

For decades, enzyme products have benefited industrial processing. As industrial producers shift to more sustainable processes requiring lower inputs of energy and chemicals, enzymes provide a simple solution to an essential need.