Preparation method of (2S,3S)-2,3-butanediol
A technology of butanediol and diacetyl, which is applied in the field of biochemistry, can solve the problems of low concentration and optical purity, low chiral purity, and low efficiency, and achieve easy-to-obtain raw materials, simple synthetic routes, and high efficiency Effect
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[0032] Example 1
[0033] This embodiment provides a method for preparing a solution containing ketoreductase, which includes the following steps:
[0034] (1) Insert the ketoreductase coding gene between the Nco I and EcoR I restriction sites of the pET28a(+) vector to obtain a recombinant vector; the nucleotide sequence of the ketoreductase coding gene is shown in the sequence table The amino acid sequence of the ketoreductase is shown in SEQ ID NO: 2 in the sequence table.
[0035] (2) Transform the aforementioned recombinant vector into E. coli BL21 to obtain a recombinant strain.
[0036] (3) Inoculate the above recombinant strain into LB solid medium containing chloramphenicol, and place it at 37°C for activation culture for 20 hours to obtain colonies; use an inoculating loop to pick a single colony and inoculate it into 50 mL of LB medium In a 250mL Erlenmeyer flask (with chloramphenicol), cover with a sealing film and fasten it with a rubber band; then, place the Erlenmeyer ...
Example Embodiment
[0038] Example 2
[0039] This embodiment provides a method for preparing (2S, 3S)-2,3-butanediol, which includes the following steps:
[0040] (1) Take 6.06 g of the ketoreductase-containing solution obtained in Example 1 above and 20 mL of phosphate buffered saline solution with a concentration of 0.01 mol / L in a clean 250 mL single-necked flask equipped with magnets, and place the single-necked flask The solution in the one-neck flask was stirred and mixed at a speed of 400 rpm in a water bath at 20°C to obtain a mixed solution.
[0041] (2) During the stirring process, keep the temperature of the water bath constant, and add 0.202g of nicotinamide adenine dinucleotide, 100mL of isopropanol and 20.2g of 2,3-butanedione to the above mixture in sequence The airtight reaction is performed to obtain a reaction liquid.
[0042] (3) Perform vacuum filtration on the above reaction solution to obtain a light yellow filtrate, and then place the filtrate under the conditions of 45° C. and 0...
Example Embodiment
[0044] Example 3
[0045] This embodiment provides a method for preparing (2S, 3S)-2,3-butanediol, which includes the following steps:
[0046] (1) Take 10.1 g of the ketoreductase-containing solution obtained in Example 1 above and 20 mL of the phosphate buffered saline solution with a concentration of 0.2 mol / L in a clean 250 mL single-necked flask containing a magnet, and place the single-necked flask The solution in the one-neck flask was stirred and mixed at a speed of 400 rpm in a water bath at 45°C to obtain a mixed solution.
[0047] (2) During the stirring process, keep the temperature of the water bath constant, and sequentially add 0.606g of nicotinamide adenine dinucleotide, 180mL of isopropanol and 20.2g of 2,3-butanedione to the above mixture The airtight reaction is performed to obtain a reaction liquid.
[0048] (3) Perform vacuum filtration of the above reaction liquid to obtain a light yellow filtrate, and then place the filtrate under the conditions of 55° C. and 0...
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