Mass preparation method of ph450 enzyme
A large-scale, enzyme-induced technology, applied in the field of protein extraction, can solve the problems of PH450 enzyme activity and quality reduction, inability to achieve high-quality application, etc., to achieve high efficiency, easy induction and separation, and good quality.
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[0017] Example of the present invention proposes a large amount of preparation method of PH450 enzyme, comprises the steps:
[0018] S10, carrying out mass culture of cyanobacteria, and inducing with PH450 enzyme inducer during the culture process;
[0019] S20, after the mass culture in step S10 is completed, the total protein of cyanobacteria cells is extracted;
[0020] S30, using human PH450 enzyme as the antigen for immune reaction to prepare polyclonal antibody;
[0021] S40, using polyclonal antibody as an affinity medium to prepare an affinity chromatography column;
[0022] S50, pass the total protein of cyanobacteria cells extracted in step S20 on the affinity chromatography column in step S40.
[0023] In the above-mentioned method of the present invention, the process of its preparation is induced and cultivated by a large number of cyanobacteria, and a large amount of cell-modified PH450 enzymes are produced in the process of cyanobacteria metabolism, which has ...
Embodiment 1
[0043] S11, configure BG-11 medium (configure according to the mother liquor composition culture recorded in the microbiological experiment manual), and obtain cyanobacteria species at the same time (the present invention preferably chooses the crystal cyanobacteria of Lake Neuchâtel and "Shuisheng No. 1042" cyanobacteria, and selects Its purpose is to be more active than other cyanobacteria in the pigment metabolism of the usual domestic "Shuisheng No. 1042", and the amount of enzyme production is higher, which is suitable for the mass production of the present invention); S12, inoculate the cyanobacteria species to BG Cultivation in -11 medium: first transfer a small amount of algae to fresh medium, the inoculation ratio is 300,000 to 2 million cells / ml medium (generally inoculate as many as possible), and use a synchronous incubator placed in a cool white fluorescent tube (200-400footcandle) for 1-2 days to observe the initial growth status, after the growth is better, move ...
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