Method for synthesizing L-tertiary leucine through double enzymes
A technology of tert-leucine and trimethylpyruvate, which is applied in the field of double-enzyme synthesis of L-tert-leucine, can solve the problems of long reaction time and high production cost, shorten the reaction time, reduce the reaction cost, and improve the reaction rate. efficiency effect
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Embodiment 1
[0013] A method for synthesizing L-tert-leucine with two enzymes, configuring reaction solution A: adding 0.5 g of trimethylpyruvate, 0.47 mL of ammonia water with a concentration of 30% and 0.5 mL of isopropanol to a concentration of 0.63 M In the ammonium formate solution with a volume of 2mL, stir evenly, and adjust the pH value to 9.0; prepare reaction solution B: mix 7.5mg leucine dehydrogenase (commercially available), 7.5mg isopropanol dehydrogenase (purchased in Ningbo Madison Pharmaceutical Technology Co., Ltd. (trade name: 111EM006) and 2.5mg of coenzyme NADH (commercially available) were dissolved in water, stirred evenly, and made into 2.03mL enzyme solution; React at a reaction temperature of 40°C and a stirring rate of 400 rpm for 4 hours. After the reaction is complete, filter to obtain a solid of L-tert-leucine. The above-mentioned reacted reaction formula is as follows: figure 1 shown.
Embodiment 2
[0015] It is basically the same as Example 1, except that the mass of trimethylpyruvate is 0.25 g, the pH value is 9.5, the reaction temperature is 25° C., and the reaction time is 8 hours.
Embodiment 3
[0017] It is basically the same as Example 1, except that the mass of trimethylpyruvate is 1.0g, the pH value is 8.0, and the reaction temperature is 30°C. The reaction solution A is poured into the reaction solution B in two times: that is, the reaction solution is first Add 1.5 mL of solution A to reaction solution B, react for 3 hours, and then add the remaining amount of reaction solution A to reaction solution B for 9 hours.
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