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An aminobutyric acid and biological preparation technology, applied in chemical recovery, fermentation and other directions, can solve the problems of increasing catalyst cost, difficult to meet the requirements of raw materials, increasing raw material cost, etc., and achieve complete conversion of raw materials, low cost, and reduced raw material cost. Effect
Active Publication Date: 2012-07-25
ENZYMEWORKS
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Problems solved by technology
However, in the method of the invention, the three enzymes need to be cloned and expressed, then the cells are crushed and freeze-dried to obtain a freeze-dried powder enzyme preparation, which increases the cost of the catalyst; the reaction system needs to use phosphate as a buffer system, which increases the cost of raw materials . The post-reaction treatment of this process is only a preliminary treatment. According to this process, impurities such as inorganic salts and ammonium formate in the product are difficult to meet the requirements of raw materials
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Embodiment 1
[0020] Take L-threonine 100mg, ammonium formate 160mg, L-threonine deaminase 0.6mg, whole cells (Applied and environmental microbiology 1997, 63(12):4651) 50mg, pyridoxal phosphate (PLP) 0.05mg , dissolved in 1mL of water, keep the temperature of the water bath at 35°C, stir at 500rpm. LC / MS tracked and detected the conversion, and the conversion rate was 98.6% within 22 hours.
Embodiment 2
[0022] Take 20 g of L-threonine, 12.7 g of ammonium formate, 0.12 g of L-threonine deaminase, 13 g of whole cells (Applied and environmental microbiology 1997, 63(12):4651), 0.01 g of PLP, and dissolve them in 200 mL of water , keep the temperature of the water bath at 35°C, stir at a speed of 500rpm. LC / MS tracked and detected the conversion, and the conversion rate was greater than 99% within 21 hours. Stop the reaction, heat the reaction solution to 70-80°C for about 1 hour to destroy the enzyme protein, and centrifuge to remove the protein. Use formic acid to adjust the pH of the supernatant to about 5.8, evaporate to dryness and recover water, and the residue is a light yellow solid. The solid is washed with 200-300mL of methanol to remove ammonium formate, and ammonium formate and solvent methanol are recovered from the washing liquid, filtered by suction, and dried to obtain The white solid was 14.2 g, and the yield was 71%. Purity greater than 99.5%, specific rotat...
Embodiment 3
[0024] Take 12 g of L-threonine, 7.56 g of ammonium formate, 0.06 g of L-threonine deaminase, 3 g of whole cells (Applied and environmental microbiology 1997, 63(12):4651), 0.005 g of PLP, and dissolve them in 100 mL of water , keep the temperature of the water bath at 35°C, stir at a speed of 500rpm. LC / MS tracked and detected the conversion, and the 24h conversion rate was greater than 99%. Yield 86%.
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Abstract
The invention relates to an L-2-reanal biological preparation method, which includes: utilizing L-threonine as raw materials, stirring the L-threonine in water at the temperature of 15-50 DEG C under the catalytic action of L-threonine deaminase and whole cells with a leucine dehydogenase and coenzyme regenerating function, and obtaining L-2-reanal after reaction. In the L-2-reanal biological preparation method, water is used for substituting for buffer salt solution to form a water-phase reaction system, cost of raw materials is reduced, threonine deaminase and whole cells used for catalyzing can be produced in batch by fermentation of microorganisms, are low in cost and wide in sources. Biological enzymatic catalysis reaction is water-phase reaction, transformation conditions of the enzymic method are mild, raw materials are transformed thoroughly, post-treatment is simple, products are separated by isoelectric point crystallization technology and film evaporation separation technology jointly, and the L-2-reanal biological preparation method is capable of recovering water and ammonium formate is low in cost, free of discharge of waste water and waste residues, environment-friendly in process and applicable to industrialized production of the L-2-reanal.
Description
technical field [0001] The invention relates to a biological preparation method of L-2-aminobutyric acid. Background technique [0002] L-2-aminobutyric acid is a natural amino acid that inhibits the transmission of human nerve information. As an important chemical raw material and pharmaceutical intermediate, it has been widely used in drug synthesis. L-2-aminobutyric acid is the main raw material for the synthesis of the new antiepileptic drug levetiracetam, and it is also the key chiral precursor for the synthesis of the antibacterial and anti-tuberculosis drug ethambutol hydrochloride; A chiral intermediate of a chiral drug. [0003] Limited by technology and cost, the output of L-2-aminobutyric acid in my country cannot meet the demand of domestic and foreign trade export. There are chemical and biological methods for the preparation of L-2-aminobutyric acid. The traditional chemical methods, whether organic synthesis or chemical resolution, lose their competitivenes...
Claims
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