What are the factors affecting the effectiveness of enzyme preparations?
In the pursuit of a green and low-carbon lifestyle, enzyme preparations have penetrated every aspect of our lives due to their characteristics of high efficiency, safety, no toxic side effects and low environmental impact. For example, enzyme preparations can be used in the bread and buns we eat, the fruit juices and drinks we drink, the seasonings we use in stir-frying, and the paper documents we use.
Therefore, understanding the chemical nature of enzyme preparations is important for their proper use.
1. The influence of pH
Each enzyme only exhibits high activity within a specific pH range, and this pH value is the optimum pH for enzyme action. Generally speaking, enzymes are most stable at the optimum pH, so the pH for enzyme action is also the pH at which they are stable. If the pH of the enzyme reaction is too high or too low, the enzyme will be irreversibly damaged, its stability and activity will decrease, and it may even become inactive. The optimal pH range of different enzymes is different, and they can be acidic, neutral or alkaline. For example, according to the optimal pH for protease action, they are often divided into acidic proteases, neutral proteases and alkaline proteases. The pH at which an enzyme acts is also a parameter measured under certain conditions. The optimal pH for enzyme action varies with temperature or substrate. The higher the temperature, the narrower the stable pH range for enzyme action. Therefore, the pH of the reaction must be strictly controlled during the enzyme catalytic reaction.
2. The effect of temperature
Under certain conditions, each enzyme has an optimum temperature for action. At this temperature, the enzyme has the highest activity, the best effect, and is relatively stable. The rate of the enzyme-catalyzed reaction increases and the loss of enzyme activity due to heat denaturation reaches an equilibrium. This temperature is the optimum temperature for enzyme action. Each enzyme has an active and stable temperature. At this temperature, under certain conditions of time, pH and enzyme concentration, the enzyme is relatively stable and does not or very rarely lose its activity. This temperature is the stable temperature of the enzyme. If the enzyme is used above the stable temperature, it will become rapidly inactive. This thermal sensitivity of the enzyme can be expressed by the critical loss temperature Tc, which refers to the temperature at which the enzyme loses half of its activity in 1 hour. Therefore, generally speaking, an enzyme can only catalyze effectively within its effective temperature range. For every 10°C increase in temperature, the enzyme reaction rate increases by 1 to 2 times. The effect of temperature on the action of an enzyme is also related to the time it is exposed to heat. As the reaction time increases, the optimum temperature for the enzyme decreases. In addition, factors such as the substrate concentration of the enzyme reaction, the type of buffer, the activator, and the purity of the enzyme can also change the optimum temperature and stability of the enzyme.
3. The influence of enzyme concentration and substrate concentration
Substrate concentration is the main factor determining the rate of an enzyme’s catalytic reaction, given a certain temperature, pH and enzyme concentration. When the substrate concentration is very low, the rate of the enzyme’s catalytic reaction increases rapidly with the substrate concentration, and the two are directly proportional. As the substrate concentration increases, the reaction rate slows down and no longer increases in direct proportion. The relationship between substrate concentration and the rate of an enzyme’s catalytic reaction can generally be expressed using Michaelis-Menten equation. Sometimes, when the substrate concentration is very high, the enzyme reaction rate may decrease due to substrate inhibition. When the substrate concentration greatly exceeds the enzyme concentration, the enzyme catalytic reaction rate is generally proportional to the enzyme concentration. In addition, if the enzyme concentration is too low, the enzyme may sometimes become inactive, preventing the reaction from proceeding. In enzyme-catalysed reactions carried out in food processing, the amount of enzyme used is generally much less than the amount of substrate, and the cost of the enzyme must also be considered.
4. The effect of inhibitors
Many substances can weaken, inhibit or even destroy the action of enzymes. These substances are called enzyme inhibitors. Examples include heavy metal ions (Fe3+, Cu2+, Hg+, Pb+, etc.), carbon monoxide, hydrogen sulfide, organic cations, ethylenediamine, and tetraacetic acid. In actual production, it is important to understand and avoid the effects of inhibitors on enzyme catalysis.
5. The effect of activators
Many substances have the effect of protecting and increasing enzyme activity, or promoting the conversion of inactive enzyme proteins into active enzymes. These substances are collectively referred to as enzyme activators. Activators can be divided into three categories: the first category is inorganic ions, such as cations such as Na+, K+, Ca2+, Mg2+, Cu2+, Co2+, and Zn2+, and anions such as Cl-, NO3-, PO43-, and SO42-. The second type is organic matter with small molecules, mainly the B vitamins and their derivatives. The third type is high-molecular substances with protein properties. Activators have an effect on the speed of enzymatic reactions similar to substrate concentration, but are rarely used in actual production.
6. The influence of the storage environment
Enzyme preparations are dormant at low temperatures. To preserve enzymes for a long time without losing activity, enzyme activity is lost at 10°C by 5-10%/6 months, and at room temperature by 10-15%/6 months. Therefore, the key is dryness and low temperature. Heat and light can easily deactivate enzymes. Therefore, enzyme preparations should be stored in airtight containers at low temperatures away from light. In addition, the higher the moisture content of the enzyme preparation, the more likely it is to become inactive. Therefore, powdered enzyme preparations are generally easier to store and transport. In addition, some metal ions can also cause enzymes to lose activity or inhibit enzyme activity. You should avoid choosing containers with metal ions to store enzyme preparations.
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