Fermentation method for high-content 12[alpha]-hydroxysteroid dehydrogenase fermentation liquor
A technology of hydroxysteroid and fermentation method, which is applied in the field of fermentation of fermented liquid with high content of 12α-hydroxysteroid dehydrogenase, can solve the problem of low activity and achieve the effect of high activity, high enzyme activity and reasonable nutrition mix
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Embodiment 1
[0029]A method for fermenting a high-content 12α-hydroxysteroid dehydrogenase fermented liquid, taking spare 12α-hydroxysteroid dehydrogenase expression gene recombinant E. Bacteria fermentation medium was cultivated, and induction medium was added after 2 hours for a total of 8 hours; the recombinant Escherichia coli expressing the 12α-hydroxysteroid dehydrogenase gene was a secondary fermentation strain, stored in glycerol tubes, and The fraction is 1-5% of the inoculum size, which is inserted into an Erlenmeyer shaker flask containing the LB liquid sterile medium, and placed in a shaker at 37°C, 40RH%, and 240rpm for 12 hours for later use; the new sterile fermentation medium, Percentage by weight, including corn flour 0.1%, corn steep liquor 2%, nitrate 0.05%, ammonium salt 0.05%, ammonia water 0.1%, phosphate 0.1%, sodium chloride 0.2%, magnesium sulfate 0.02%, zinc sulfate 0.02%, 0.05% defoamer, the balance is water; the induction medium, by weight percentage, includes 0...
Embodiment 2
[0031] A method for fermenting a high-content 12α-hydroxysteroid dehydrogenase fermented liquid, taking spare 12α-hydroxysteroid dehydrogenase expression gene recombinant E. Bacteria fermentation medium was cultivated, and induction medium was added after 2 hours for a total of 8 hours; the recombinant Escherichia coli expressing the 12α-hydroxysteroid dehydrogenase gene was a secondary fermentation strain, stored in glycerol tubes, and The fraction is 1-5% of the inoculum size, which is inserted into an Erlenmeyer shaker flask containing the LB liquid sterile medium, and placed in a shaker at 37°C, 45RH%, and 240rpm for 12 hours for later use; the new sterile fermentation medium, Percentage by weight, including corn flour 0.3%, corn steep liquor 4%, nitrate 0.075%, ammonium salt 0.075%, ammonia water 0.45%, phosphate 0.3%, sodium chloride 0.15%, magnesium sulfate 0.015%, zinc sulfate 0.015%, 0.05% defoamer, the balance is water; the induction medium, by weight percentage, inc...
Embodiment 3
[0033] A method for fermenting a high-content 12α-hydroxysteroid dehydrogenase fermented liquid, taking spare 12α-hydroxysteroid dehydrogenase expression gene recombinant E. Bacteria fermentation medium was cultivated, and induction medium was added after 2 hours for a total of 8 hours; the recombinant Escherichia coli expressing the 12α-hydroxysteroid dehydrogenase gene was a secondary fermentation strain, stored in glycerol tubes, and The fraction is 1-5% of the inoculum size, which is inserted into an Erlenmeyer shaker flask containing the LB liquid sterile medium, and placed in a shaking table at 37°C, 50RH%, and 240rpm for 12 hours for later use; the new sterile fermentation medium, Percentage by weight, including 0.5% corn flour, 6% corn steep liquor, 0.2% nitrate, 0.2% ammonium salt, 1% ammonia water, 0.5% phosphate, 0.5% sodium chloride, 0.05% magnesium sulfate, 0.05% zinc sulfate, 0.05% defoamer, the balance is water; the induction medium, by weight percentage, includ...
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