Continuous enzymatic method for producing L-tert-leucine

A technology of tert-leucine and enzymatic method, which is applied in the field of splitting and preparing L-tert-leucine by using aminoacylase, can solve the problems of large solvent consumption, low production efficiency and complexity, and achieve high raw material utilization rate , reduced usage, low production cost

Active Publication Date: 2015-07-15
瑞博(杭州)医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the aforementioned methods all adopt batch reaction, and the production efficiency is low; the reaction process needs to constantly adjust the pH, and the process is loaded down with trivial details; after the reaction is finished, the ineffective N-phenylacetyl-D-tert-leucine and phenylacetic acid are directly acidified and extracted, and the solvent consumption is large. , and the two substances are not easy to separate and utilize; the invalid body has not been utilized or the condition is complicated although there is racemization treatment

Method used

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  • Continuous enzymatic method for producing L-tert-leucine
  • Continuous enzymatic method for producing L-tert-leucine
  • Continuous enzymatic method for producing L-tert-leucine

Examples

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Effect test

Embodiment 1

[0033] Add 200g of N-phenylacetyl-DL-tert-leucine into 2000mL of pure water, and fully dissolve 165mL of 5M sodium hydroxide to prepare a substrate solution with a substrate concentration of 9.24% and a pH of 7.0. The product solution was passed through the Bacillus megaterium penicillin acylase reaction column at a flow rate of 1000mL / h, and the reaction was carried out at a room temperature of about 25°C. The conversion rate was determined to be 16.4% by sampling after 1 hour of reaction, and 49.7% after reaction 6 hours. Collect the reaction solution, and replace the remaining reaction solution in the column with 200mL pure water to finally obtain 2200mL of the reaction solution containing L-tert-leucine, add 166mL of 5M hydrochloric acid dropwise, filter under reduced pressure to obtain a white precipitated solid, and dry it in an oven at 60°C. Weighing 103.2g, the unreacted substrate content is 91.3%, which is N-phenylacetyl-D-tert-leucine. Take 1g of solid methanol and di...

Embodiment 2

[0035] Add 400g of N-phenylacetyl-DL-tert-leucine into 2000mL of pure water, and fully dissolve 331mL of 5M sodium hydroxide to prepare a substrate solution with a substrate concentration of 17.16% and a pH of 7.0. The product solution passed through the immobilized Amano aminoacylase reaction column at a flow rate of 500mL / h, and the reaction was carried out at a room temperature of about 25°C. After 2 hours of reaction, the conversion rate was measured to be 12.7%, and after 12 hours of reaction, the conversion rate was analyzed to be 48.3%. Collect the reaction solution, and replace the remaining reaction solution in the column with 200mL pure water to finally obtain 2350mL of the reaction solution containing L-tert-leucine, add 333mL of 5M hydrochloric acid dropwise, filter under reduced pressure to obtain a precipitated solid, dry it in an oven at 60°C, weigh The weight is 210.8g, which is 89.7% of the unreacted substrate content, mainly N-phenylacetyl-D-tert-leucine and t...

Embodiment 3

[0037] Add 300 g of the substrate obtained by racemization of Example 1 and Example 2 (N-phenylacetyl-DL-tert-leucine content 90.2%, the rest is phenylacetic acid) into 2000 mL of pure water, fully dissolve 275 mL of 5M sodium hydroxide, and prepare To form a substrate solution with a substrate concentration of 11.89% and a pH of 7.0, use a constant flow pump to pass the substrate solution through the Alcaligenes faecalis penicillin acylase reaction column at a flow rate of 1000mL / h, and react at a room temperature of about 25°C for 3 hours The conversion rate was determined to be 28.3% by sampling, and the conversion rate was analyzed after 7 hours of reaction to be 48.3%. Collect the reaction solution, and replace the remaining reaction solution in the column with 200mL pure water to finally obtain 2300mL of the reaction solution containing L-tert-leucine, add 279mL of 5M hydrochloric acid dropwise, filter under reduced pressure to obtain a white precipitated solid, and dry i...

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Abstract

The invention relates to a method for preparing an optical isomer by continuously resolving a nitrogenous organic compound raceme with an enzyme, and particularly relates to a method for preparing L-tert-leucine by resolution with an aminoacylase, specifically a continuous enzymatic method for producing L-tert-leucine. The method comprises the following steps: by using N-phenylacetyl-DL-tert-leucine and an inorganic alkali as raw materials and pure water as a reaction medium, preparing a substrate solution with a certain concentration; passing the substrate solution through an aminoacylase reaction column at a certain flow rate; under the enzyme catalytic actions, selectively hydrolyzing N-phenylacetyl-L-tert-leucine to generate L-tert-leucine and phenylacetic acid; and after the reaction finishes, acidifying the reaction solution, carrying out vacuum filtration to separate out unreacted N-phenylacetyl-D-tert-leucine, and separating and extracting phenylacetic acid from the filtrate with an organic solvent, wherein the L-tert-leucine is in the water phase, and the unavailable N-phenylacetyl-D-tert-leucine is racemized by high-temperature reflux and put into the resolving reaction again.

Description

technical field [0001] The invention relates to a method for splitting racemates of nitrogen-containing organic compounds by enzymes to prepare optical isomers, in particular to a method for splitting and preparing L-tert-leucine by using aminoacylases. Background technique [0002] L-tert-leucine, English name L-tert-Leucine, chemical name L-2-amino-3,3-dimethylbutyric acid, is a compound with the following structure, [0003] [0004] L-tert-leucine [0005] L-tert-leucine is a non-proteinogenic chiral amino acid and an important pharmaceutical intermediate, mainly used in the preparation of anti-cancer and anti-AIDS (HIV) drugs. Currently, there are pharmaceuticals such as Bristol-Myers Squibb, Pfizer, and Abbott The new protease inhibitor anti-HIV drugs listed by the company all use this product as an intermediate, such as the anti-HIV drug Atazanavir (Atazanavir), whose global sales in 2006 reached 3.226 billion US dollars, and there are six other anti-HIV drugs in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P13/06
Inventor 罗积杏壮晓健沈文和车大庆
Owner 瑞博(杭州)医药科技有限公司
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