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Non-mechanical method of cell disruption technology
Cell disruption techniques have been developed to accommodate different uses and different types of cell wall disruption. The crushing method can be classified into two major categories: mechanical methods and non-mechanical methods. The following is a classification of non-mechanical methods:
1: chemical crushing method
Chemical treatment can dissolve cells or extract intracellular components. Chemical reagents such as acids, bases, surfactants and organic solvents
2: detergent crushing method
Protein renaturation utilizes the density difference between the inclusion body and the cell debris, and separates the inclusion body from the cell debris and the soluble protein by centrifugation to obtain a clean inclusion body, and then lynates the inclusion body. In this way, a large amount of impurities such as impurities, nucleic acids, pyrogens and endotoxins are first removed, so that the subsequent separation and purification is simple. From this perspective, the formation of inclusion bodies is also beneficial for separation and purification.
3: Enzymatic solution breaking method
Enzymatic hydrolysis is the treatment of bacterial cells by the enzyme that dissolves the cell wall, and the cell wall is partially or completely destroyed, and then the cell membrane is destroyed by osmotic pressure shock or the like to further increase the permeability of the intracellular product. Lysozyme is suitable for the decomposition of Gram-negative cells. When applied to Gram-positive bacteria, it needs to be supplemented with EDTA to make it more effective on the cell wall. The cell wall of eukaryotic cells differs from prokaryotic cells in that different enzymes are required.
4: osmotic shock crushing method
Osmotic shock is a milder method of crushing cells. The cells are placed in a solution with high osmotic pressure (such as a certain concentration of glycerol or sucrose solution). Due to the osmotic pressure, the water in the cells exudes outward and the cells occur. Shrinkage, when equilibrium is reached, the medium is rapidly diluted, or the cells are transferred to water or buffer. Due to the sudden change in osmotic pressure, extracellular water rapidly infiltrates into the cells, causing the cells to rapidly expand and rupture.
5: freeze-thaw fracture method
The cells were frozen at a low temperature (about -15 ° C), then thawed at room temperature, and repeatedly broken to achieve a wall breaking effect. Due to freezing, on the one hand, the hydrophobic bond structure of the cell membrane can be broken, thereby increasing the hydrophilic property of the cell, and on the other hand, the intracellular water crystallizes to form ice crystal grains, causing the cells to swell and rupture. This method can be used for cells with weak cell walls.