Biotransformation method for L-tert-leucine

A technology of tertiary leucine and biotransformation, applied in the field of preparation of unnatural amino acids, can solve the problems of inability to meet industrial production, no clear post-extraction process, decrease in reaction rate, etc. Feed reduction effect

Inactive Publication Date: 2018-09-25
南宁信肽生物技术有限责任公司
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Several patent applications have been disclosed for this route, and the main problems of its existence are: in the currently disclosed patents, the initial substrate concentration of the reaction is 10-15%, and L-tert-leucine dehydrogenase has a high substrate concentration. It will be significantly inhibited by the substrate, and the reaction rate will drop sharply. Under the condition of high ammonia concentration, the excess substrate and the product accumulation will form by-products, indicating that the method is difficult to repeat.
In addition, the dosage of the enzyme is too much, the dosage of the expensive coenzyme NAD reaches 1% (w / w) of the substrate, the economy is poor, and there is no clear post-extraction process, which still cannot meet the needs of industrial production

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Biotransformation method for L-tert-leucine
  • Biotransformation method for L-tert-leucine
  • Biotransformation method for L-tert-leucine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] The preparation process of L-tert-leucine crystal comprises the following steps:

[0058] 1) Heat 8 parts by weight of compound glue in 50 parts by weight of distilled water to dissolve at 80°C. The compound glue includes 0.3 part by weight of calcium alginate, 0.9 part by weight of potassium alginate, 0.4 part by weight of sodium alginate and 6.4 parts by weight of poly Ethylene glycol, after cooling down to 40°C, add 1 part by weight of leucine dehydrogenase and 5.5 parts by weight of formate dehydrogenase, stir evenly, then store in the refrigerator at 4°C, let it cool and solidify, and then it will be fixed oxidized leucine dehydrogenase, formate dehydrogenase. Cut the immobilized leucine dehydrogenase and formate dehydrogenase into 1×1cm 3 cubes, spare.

[0059] 2) taking dichloropinatone as raw material to prepare sodium trimethylpyruvate solution;

[0060] Preparation of sodium trimethylpyruvate solution specifically comprises the steps:

[0061] a. Add 1 wei...

Embodiment 2

[0067] The preparation process of L-tert-leucine crystal comprises the following steps:

[0068] 1) Heat 7 parts by weight of carrageenan in 42 parts by weight of distilled water to 85°C to dissolve, add 1 part by weight of leucine dehydrogenase and 6 parts by weight of formate dehydrogenase after cooling down to 35°C, stir evenly, and place Store in the refrigerator at 4°C, let it cool and solidify, and then obtain immobilized leucine dehydrogenase and formate dehydrogenase. Cut the immobilized leucine dehydrogenase and formate dehydrogenase into 1×1cm 3 cubes, spare.

[0069] 2) Prepare sodium trimethylpyruvate solution with dichloropinatone as raw material; the preparation method is as in Example 1.

[0070] 3) Using sodium trimethylpyruvate solution as a substrate, using coenzyme NAD+ and immobilized leucine dehydrogenase and formate dehydrogenase as biocatalysts, adding ammonium formate to establish a transformation system for biotransformation reaction, and obtaining a...

Embodiment 3

[0074] The preparation process of L-tert-leucine crystal comprises the steps:

[0075] 1) Heat 9 parts by weight of compound glue in 72 parts by weight of distilled water to dissolve at 88°C. Ethylene glycol, after cooling down to 45°C, add 1 part by weight of leucine dehydrogenase and 4 parts by weight of formate dehydrogenase, stir evenly, then store in the refrigerator at 4°C, let it cool and solidify, and then it will be fixed. oxidized leucine dehydrogenase, formate dehydrogenase. Cut the immobilized leucine dehydrogenase and formate dehydrogenase into 1×1cm 3 cubes, spare.

[0076] 2) Prepare sodium trimethylpyruvate solution with dichloropinatone as raw material; the preparation method is as in Example 1.

[0077]3) Using sodium trimethylpyruvate solution as a substrate, using coenzyme NAD+ and immobilized leucine dehydrogenase and formate dehydrogenase as biocatalysts, adding ammonium formate to establish a transformation system for biotransformation reaction, and o...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
purityaaaaaaaaaa
optical purityaaaaaaaaaa
optical purityaaaaaaaaaa
Login to view more

Abstract

The invention discloses a biotransformation method for L-tert-leucine. The method includes the following steps: establishing a transformation system by utilizing sodium trimethylpyruvate as a substrate, utilizing coenzyme NAD+ and immobilized leucine dehydrogenase and formate dehydrogenase as a biocatalyst and by adding an ammonium salt; and performing a biotransformation reaction to obtain a transformation liquid containing the L-tert-leucine. The biotransformation method utilizes the immobilized leucine dehydrogenase and formate dehydrogenase as the biocatalyst which can still have enzyme activity after ten times of reuse, and an expensive coenzyme and enzyme can be recycled, so that the production cost is saved; a self-made sodium trimethylpyruvate solution is utilized as the substrate,so that the preparation process is simplified, the production cost is saved, and purification is convenient; and the purity of a prepared L-tert-leucine crystal reaches 98.5% or above, and the optical purity is 99% or above. Process flow is simple, no special requirements are for equipment, and industrialization production is suitable. HPLC-MS and HPLC are utilized to monitor until the substrateis completely utilized.

Description

technical field [0001] The invention relates to a preparation method of unnatural amino acid, in particular to a biotransformation method of L-tert-leucine. Background technique [0002] Unnatural amino acids and their derivatives are important chemical intermediates. Among them, L-tert-leucine is an important unnatural amino acid, which can be used to synthesize pharmaceutical intermediates, and as a chiral inducer for asymmetric synthesis and unnatural A resolution reagent for symmetrical resolution, widely used in the synthesis of biological inhibitors, bioactive peptides, anticancer and antiviral drugs. At present, the protease inhibitor anti-HIV new drugs listed by pharmaceutical companies such as Bristol-Myers Squibb, Pfizer, and Abbott all use L-tertiary leucine as an intermediate, such as the anti-HIV drug Atazanavir (Atazanavir), its global sales in 2006 In addition, six new anti-HIV and anti-tumor drugs in clinical trials also use L-tert-leucine as an intermediate...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/04C12N11/10C12N11/08C07C51/373C07C59/185
CPCC12P13/04C07C51/373C07C51/41C12N9/0008C12N9/0016C12N11/08C12N11/10C12Y102/01002C12Y104/01009C07C59/185C07C59/01
Inventor 胡磊潘崇德陈迈陈建华
Owner 南宁信肽生物技术有限责任公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap