Preparation of Tetramethylpyrazine
A technology of ligustrazine and natto subtilis, applied in the field of bioengineering, can solve safety problems, environmental pollution and other problems, and achieve the effects of high safety, no environmental pollution, and simple operation process
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Embodiment 1
[0031] The LB slant seeds of Bacillus subtilis natto were cultured for 24 hours, then transferred to LB liquid medium, and cultivated in a shaker at 37° C. and 150 r / min for 18 hours to obtain a seed culture solution.
[0032] Weigh 100 g of soybeans soaked in running water and add them to a 500 mL Erlenmeyer flask, and sterilize at 115°C for 25 minutes to obtain the required culture medium.
[0033] Aseptically insert 2g of seed culture solution into the above medium, and after 144 hours of transformation at 37°C, 102g of a product containing ligustrazine is obtained. Add 510mL of chloroform, and reflux extraction at 30°C for 1.5 hours under ultrasonic treatment with a power of 550W. Finally, filter to remove the residue to obtain an extract, add 0.2 mol / L dilute hydrochloric acid with the same volume as the extract, mix evenly, and leave to separate for stratification. The aqueous phase is the hydrochloridized Ligustrazine aqueous solution.
[0034] The above hydrochloridize...
Embodiment 2
[0036] The LB slant seeds of Bacillus subtilis natto were cultured for 24 hours, then transferred to LB liquid medium, and cultivated in a shaker at 37° C. and 150 r / min for 18 hours to obtain a seed culture solution.
[0037] Weigh 100 g of soybeans soaked in running water and add them to a 500 mL Erlenmeyer flask, and sterilize at 115° C. for 25 minutes to obtain the required culture medium.
[0038]Insert 2g of seed culture liquid into the above medium aseptically, and carry out culture transformation at 37°C. After 96 hours of culture transformation, add 0.2g of acetoin. After 144 hours of culture transformation, 102.2g of the product containing ligustrazine is obtained, and add 511mL of acetone , under ultrasonic-assisted treatment with a power of 550W, reflux extraction at 43°C for 1.5 hours and then filter to remove the residue to obtain the extract, add 0.2mol / L dilute hydrochloric acid with the same volume as the extract, mix well and then let it stand for stratificati...
Embodiment 3
[0041] The LB slant seeds of Bacillus subtilis natto were cultured for 24 hours, then transferred to LB liquid medium, and cultivated in a shaker at 37° C. and 150 r / min for 18 hours to obtain a seed culture solution.
[0042] Weigh 100 g of soybeans soaked in running water and add them to a 500 mL Erlenmeyer flask, and sterilize at 115° C. for 25 minutes to obtain the required culture medium.
[0043] Aseptically insert 5g of seed culture liquid into the above medium, and carry out culture transformation at 37°C. After 96 hours of culture transformation, add 0.2g of acetoin. After 144 hours of culture transformation, 105.2g of products containing ligustrazine are obtained. Add 5260mL Chloroform, under ultrasonic treatment with a power of 880W, reflux extraction at 25°C for 2.5 hours, then filter to remove the residue to obtain the extract, add 0.2mol / L dilute hydrochloric acid with the same volume as the extract, mix well, and then let it stand for stratification , the aqueou...
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