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Method for preparing levodopa with one-pot enzymatic method

A technology for levodopa and enzymatic preparation, which is applied in the field of bioengineering, can solve the problems of serious environmental pollution and high production cost, and achieves the effects of high yield, low production cost and low environmental protection pressure.

Active Publication Date: 2018-01-16
CHANGXING PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, commercially available pyruvate or L-tyrosine is basically chemically synthesized, and the production cost is relatively high, and the environmental pollution is particularly serious.

Method used

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  • Method for preparing levodopa with one-pot enzymatic method
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  • Method for preparing levodopa with one-pot enzymatic method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1: the conversion of 2.5 tons of systems

[0072] Add 1600L of water, 120kg of glycine, and 55kg of formaldehyde solution (37%) in the bioreactor, and use concentrated ammonia to control the pH at 6.0 to 8.0; add 0.33kg of pyridoxal 5-phosphate, and use concentrated ammonia to control the pH At 6.0-8.0, the temperature is controlled at 32°C-38°C. Add 25kg of catechol and 650kg of enzyme solution containing recombinant D-threonine aldolase, recombinant D-serine dehydratase and recombinant tyrosine phenolase to start the reaction. During the reaction process, 37% formaldehyde solution was added to maintain the reaction pH between 6.0 and 8.0. After that, 5kg of substrate catechol was added every 10min. The total input amount of catechol is 176kg. After the catechol feeding was finished, the reaction was continued for 2 hours, and the sample was taken for detection. The concentration of levodopa was 67g / L, and the conversion rate was about 53.2%. After post-...

Embodiment 2

[0073] Embodiment 2: the transformation of 2.5 tons system

[0074]Add 1600L of water, 140kg of glycine, 62.5kg of formaldehyde solution (37%) to the bioreactor, and control the pH at 6.0 to 8.0 with concentrated ammonia water; then add 5kg of sodium sulfite, 2.5kg of EDTA, 50kg of ammonium acetate, 5 -Pyridoxal phosphate 0.33kg, again use concentrated ammonia water to control the pH at 6.0-8.0, and control the temperature at 32°C-38°C. Add 25kg of catechol and 600kg of enzyme solution containing recombinant D-threonine aldolase, recombinant D-serine dehydratase and recombinant tyrosine phenolase to start the reaction. During the reaction process, 37% formaldehyde solution was added to maintain the reaction pH between 6.0 and 8.0. After that, 5kg of substrate catechol was added every 10min. The total input amount of catechol is 205kg. After the catechol feeding was finished, the reaction was continued for 1 hour, and the sample was taken for detection. The concentration of ...

Embodiment 3

[0075] Embodiment 3: the transformation of 7.5 tons system

[0076] Add 4800L of water, 450kg of glycine, and 180kg of formaldehyde solution (37%) to the bioreactor in sequence, and control the pH at 6.0 to 8.0 with concentrated ammonia water; then add 15kg of VC, 7.5kg of EDTA, 120kg of ammonium formate, Pyridoxal 1kg, again use concentrated ammonia water to control the pH at 6.0-8.0, and control the temperature at 32°C-38°C. Add 75kg of catechol and 1800kg of enzyme solution containing recombinant L-threonine aldolase, recombinant L-serine dehydratase and recombinant tyrosine phenolase to start the reaction. During the reaction process, 37% formaldehyde solution was added to maintain the reaction pH between 6.0 and 8.0. After that, 15kg of substrate catechol was added every 10min. The total input amount of catechol is 660kg. After the catechol feeding was finished, the reaction was continued for 1.5h, and the sample was taken for detection. The concentration of levodopa w...

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Abstract

The invention discloses a method for preparing levodopa with a one-pot enzymatic method. The method is characterized in that formaldehyde, glycine and catechol are taken as substrates; formaldehyde and glycine are subjected to a catalytic reaction in the presence of aldolase and other enzymes to be converted into serine, then, serine is converted into pyruvic acid and ammonia under the action of anhydrase, and finally, pyruvic acid and catechol react under the action of tyrosine phenol lyase to produce levodopa. The method adopts novel catalysis routh design and has the advantages of being simple to operate, short in production cycle, low in production cost, high in yield, low in environmental protection pressure, suitable for mass industrial production and the like.

Description

technical field [0001] The invention relates to a biological preparation method of levodopa, in particular to a one-pot enzymatic method for preparing levodopa, which belongs to the field of bioengineering. Background technique [0002] Levodopa (3,4-dihydroxylphenylalanine, Levodopa), also known as L-dopa, chemical name is 3,4-dihydroxyphenylalanine, CAS: 59-92-7, white crystalline powder, no Odorless and tasteless; it is an important biologically active substance in organisms and an important intermediate product in the biochemical metabolic pathway from L-tyrosine to catechol or melanin. Its structural formula is as follows: [0003] [0004] Levodopa is the first-line drug for the treatment of Parkinson's disease. Although there will be some side effects after long-term use, more than 40 years have passed, and there is still no new alternative drug. Most of the drugs currently on the market are chemically synthesized. The synthesis process requires a large amount of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/22
Inventor 杨卫华钱敏帆肖延铭谈聪严燕兵
Owner CHANGXING PHARMA
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