Identify the most cost effective enzyme purification method and provide brief explanations for this method. ▪Repeated freezing and thawing ▪Sonication ▪Homogenization ▪Homogenization ▪Permeabilization lysozyme) by high pressure (French press) by grinding (bead mill) by detergents (e.g. Triton X-100) and/or enzymes (e.g.
Enzyme kinetics
In biochemistry, enzymes are proteins that act as biological catalysts. Catalysis is the addition of a catalyst to a chemical reaction to speed up the pace of the reaction. Catalysis can be categorized as either homogeneous or heterogeneous, depending on whether the catalysts are distributed in the same phase as that of the reactants. Enzymes are an essential part of the cell because, without them, many organic processes would slow down and thus will affect the processes that are important for cell survival and sustenance.
Regulation of Enzymes
A substance that acts as a catalyst to regulate the reaction rate in the living organism's metabolic pathways without itself getting altered is an enzyme. Most of the biological reactions and metabolic pathways in the living systems are carried out by enzymes. They are specific for their works and work in particular conditions. It maintains the best possible rate of reaction in the most stable state. The enzymes have distinct properties as they can proceed with the reaction in any direction, their particular binding sites, pH specificity, temperature specificity required in very few amounts.
- Enzymes need to be purified from cell extract to remove the undesired compounds which helps to enhance the enzyme stability, purity and activity.
- There are different methods used for enzyme purification based on their size, their ionic properties and on other characteristics.
- Repeated cycles of freezing and thawing helps to lyse the cell and release of its cellular contents. Enzymes can be purified by this method but it may cause damage to enzymes by the formation of ice crystals.
- In sonication, Ultrasonic energy is used to break the cell. This procedure produces heat which may damage the enzymes so it is advised to use sonication under cooling condition.
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