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How do biochemists isolate and study individual enzymes?
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Biochemistry
In biochemistry, researchers study the chemical substances and processes that occur in living organisms and cells. In particular, various processes by which cells gain energy, such as epinephrine-stimulated cAMP synthesis, have been deduced through the study of biochemistry.
Biochemists isolate and study individual enzymes by exploiting their unique physical and chemical properties to separate them from complex biological mixtures.
Separation Techniques
- Centrifugation: By spinning cell homogenates at high speeds, researchers use centrifuges to separate cellular components based on density and weight. Ultracentrifugation is specifically used to isolate proteins and determine their molecular weights.12
- Electrophoresis: This method separates molecules based on their electrical charge. Enzymes migrate at different rates through a stationary medium—such as acrylamide gels—when placed in an electric field, allowing for isolation and purity assessment.12
- Chromatography: This technique separates components based on their distribution between a mobile and a stationary phase. Gel chromatography is particularly effective for enzymes, as it separates them by molecular size; larger molecules pass through a porous bed more quickly than smaller ones.34
Study and Analysis
Once isolated, enzymes are studied to determine their structure and function. Researchers can elucidate the sequence of amino acids within an enzyme and verify its identity through tests of its catalytic activity. Kinetic studies, often using the Michaelis-Menten equation, help biochemists understand how enzymes interact with substrates and determine their efficiency.56