Techniques and Methodologies for Extracting Proteins and Purifying Enzyme X
“Enzyme X is a highly pigmented protein that imparts the characteristic color to certain blue-green algae. It also facilitates a reaction necessary to the survival of this species; we can follow the kinetics of this reaction by measuring the conversion of Substance X to Substance Y at various times during purification.”
Techniques and methodologies for extracting the proteins and purifying Enzyme X are as follows:
The blue-green tinted proteins in Enzyme X give a distinct color to algae. The proteins are contained in the cells, so the cells need to be lysed (disintegration of the cell wall or membrane.) Substance X contains Enzyme X. As a note, the compound needs to be perfectly pure before studying or it will not produce wanted results.
1. The lysis of the cells in Substance X can be achieved by a variety of methods, such as repeated freeze and thaw cycles in a freezer, causing the cell wall to rupture, spilling the contents into the surrounding solution. This method works in tough organisms. Other techniques can be utilized as well, such as the use of grinding or blending of animal or plant cells to lyse, introduction of high-frequency sound waves, or enzymatic hydrolysis followed by a detergent wash.
2. Once the cells of Substance X are lysed, the solution needs to be centrifuged using a standard laboratory centrifuge and tubes. Centrifuges work by using the sedimentation principle. Said principle is when centripetal acceleration is used to separate the lower and higher dense particles in a substance.
3. Once centrifuged, the supernatant is to be removed and labeled accordingly in a separate vessel. Supernatant is the liquid above the sediment after a solution has been run through a centrifuge. The now centrifuged solution should be ran through a device that achieves column chromatography, such as an automated fraction collector. These devices sort a solution based on particle size. Other methods to extract proteins (which can be many different
Techniques and methodologies for extracting the proteins and purifying Enzyme X are as follows:
The blue-green tinted proteins in Enzyme X give a distinct color to algae. The proteins are contained in the cells, so the cells need to be lysed (disintegration of the cell wall or membrane.) Substance X contains Enzyme X. As a note, the compound needs to be perfectly pure before studying or it will not produce wanted results.
1. The lysis of the cells in Substance X can be achieved by a variety of methods, such as repeated freeze and thaw cycles in a freezer, causing the cell wall to rupture, spilling the contents into the surrounding solution. This method works in tough organisms. Other techniques can be utilized as well, such as the use of grinding or blending of animal or plant cells to lyse, introduction of high-frequency sound waves, or enzymatic hydrolysis followed by a detergent wash.
2. Once the cells of Substance X are lysed, the solution needs to be centrifuged using a standard laboratory centrifuge and tubes. Centrifuges work by using the sedimentation principle. Said principle is when centripetal acceleration is used to separate the lower and higher dense particles in a substance.
3. Once centrifuged, the supernatant is to be removed and labeled accordingly in a separate vessel. Supernatant is the liquid above the sediment after a solution has been run through a centrifuge. The now centrifuged solution should be ran through a device that achieves column chromatography, such as an automated fraction collector. These devices sort a solution based on particle size. Other methods to extract proteins (which can be many different