Enzyme extraction and separation purification.
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Last Update: 2020-10-19
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Source: Internet
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Author: User
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(i) cell fragmentation treatment
many enzymes are present in cells. In order to extract these cytoenzymes, cells need to be crushed first. There are many methods of cell crushing, including mechanical crushing, physical crushing, chemical crushing and enzymatic crushing.
mechanical crushing method refers to the use of mashers, grinders or homogenizers to break cells. Physical crushing refers to the use of temperature difference, pressure difference or ultrasonic, such as the use of cells broken. Chemical crushing refers to the use of formaldehyde, acetone and other
organic
solvents or
s surfactants
acting on the cell membrane, so that the structure of the cell membrane is damaged or permeable changes. Enzymatic fragmentation refers to the selection of suitable enzymes, so that the cell wall is destroyed, and then in a low-permeable solution to break the protosurfacies.
(ii) Extraction of
ase refers to the process under certain conditions in which the enzyme-containing raw material after cell fragmentation is treated with an appropriate solvent, so that the enzyme is fully dissolved into the extract. Enzyme extraction methods have column method, salt solution extraction method, alkali solution extraction method and organic solvent extraction method. In order to improve the extraction rate of enzymes and prevent denaturation and infestion after enzyme extraction, care must be paid to maintaining suitable temperature and pH, and the appropriate amount of protective agents must be added.
(iii) The isolation
extract contains a variety of enzymes, in order to separate purify an enzyme from the extract, it is necessary to choose a suitable separation purification method according to the characteristics of this enzyme. Enzyme separation purification methods are many, can use the size of the enzyme relative to the molecular mass of separation purification, such as by dialysis method to separate enzymes.
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