Enzyme extraction, separation, purification and production methods

Enzyme extraction, isolation and purification

(1) Cell disruption treatment
Many enzymes are found within cells. In order to extract these intracellular enzymes, cells first need to be disrupted. There are many methods of cell disruption, including mechanical disruption, physical disruption, chemical disruption, and enzymatic disruption. Mechanical disruption method refers to using a masher, grinder or homogenizer to disrupt cells. Physical disruption method refers to the use of temperature difference, pressure difference or ultrasonic waves to disrupt cells. The chemical disruption method refers to the use of organic solvents or surfactants such as formaldehyde and acetone to act on the cell membrane to destroy the structure of the cell membrane or change its permeability. The enzymatic disruption method refers to the use of appropriate enzymes to destroy the cell wall and then disrupt the protoplasts in a hypotonic solution.

(2) Extraction
Enzyme extraction refers to the process of treating the enzyme-containing raw materials after cell disruption with an appropriate solvent under certain conditions to fully dissolve the enzyme into the extraction solution. Enzyme extraction methods include column method, salt solution extraction method, alkali solution extraction method and organic solvent extraction method. In order to improve the extraction rate of the enzyme and prevent denaturation and inactivation of the enzyme after extraction, care must be taken to maintain appropriate temperature and pH value during the extraction process, and an appropriate amount of protective agent must be added.

(3) Separation
The extract contains a variety of enzymes. If you want to separate and purify a certain enzyme from the extract, you must choose an appropriate separation and purification method based on the characteristics of the enzyme. There are many methods for enzyme separation and purification. The relative molecular mass of the enzyme can be used for separation and purification, such as separation of enzymes through dialysis.
Enzyme production methods

Enzyme production refers to a process that is pre-designed and controlled manually to obtain the required enzymes. In summary, there are three methods for enzyme production: extraction, fermentation and chemical synthesis.

(1) Extraction method
The extraction method was the earliest method adopted and is still used today. Extraction methods use various techniques to extract enzymes directly from cells or tissues of animals, plants or microorganisms. Although the extraction method is simple and easy to implement, sufficient raw materials must be available, which limits the widespread application of the extraction method. However, in areas rich in animal, plant or microbial resources, the extraction method still has application value. For example, in slaughterhouses, pancreatin can be extracted from the pancreas of livestock; in fruit processing plants, bromelain can be extracted from pineapple peels.

(2) Fermentation method
Fermentation has been the main method of producing enzymes since the 1950s. The fermentation method mainly obtains the enzymes needed by people through microbial fermentation. Fermentation methods generally include solid fermentation, liquid submerged fermentation, immobilized cell fermentation, and protoplast fermentation.

(3) Chemical synthesis method
Chemical synthesis is a technology for producing enzymes that emerged in the late 1960s. In 1969, American scientists used chemical synthesis for the first time to obtain ribonuclease containing 124 amino acids. However, the cost of chemical synthesis is relatively high, and only enzymes with known chemical structures can be synthesized. Therefore, chemical synthesis methods are still at the stage of laboratory synthesis.