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Preparation method of polypeptide synthesized amino acid protective agent and DCC product precipitate

A technology for polypeptide synthesis and protective agent, applied in the field of preparation of amino acid protective agent and DCC product precipitation, can solve the problems of difficult precipitation and separation, complex molecular structure, lack of bicyclic imine, etc., and achieves easy precipitation, low price and high precision. Effect

Inactive Publication Date: 2022-04-15
柳春曦
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, the molecular structure of the amino acid protective agent used in the polypeptide synthesis technology is complex, and the pollution is relatively large, and it is not easy to precipitate and separate. There is an urgent need for an amino acid protective agent with simple molecules and easy separation to meet market demand. DCC catalysts are added to the synthesis of amino acid chains. A small part of the product dicyclohexylurea is soluble in water and cannot be completely removed. There is a lack of a method that can remove dicycloimine dissolved in water to save costs

Method used

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  • Preparation method of polypeptide synthesized amino acid protective agent and DCC product precipitate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] A preparation method of an amino acid protective agent synthesized by a polypeptide, comprising the following steps:

[0018] Step 1: Take the protective agent X, X is sodium 4-aminobenzenesulfonate, take 1mmol of the protective agent and dissolve it in 10ml of water, fully dissolve, then take 1mmol of alanine to react, the amino group of X combines with the carboxyl end of alanine to form an amide key;

[0019] Step 2: A, B, C, D, E and other amino acids are sequentially connected to the amino terminal of alanine, and at the same time, 1 mmol of DCC carbodiimide is added to react for 2-2.5 hours until the end of the polypeptide chain synthesis;

[0020] Step 3: Take 1 mmol of alanine aminopeptidase, cut off X-alanine with an enzyme, and then add a precipitating agent calcium chloride to react, and remove the protective agent X molecule formed by the precipitation of benzenesulfonic acid group and calcium ion Ca 2+ -X-Ala-NH 2 ;

[0021] A method for preparing the D...

Embodiment 2

[0027] A method for preparing the DCC product precipitation of polypeptide synthesis, comprising the following steps:

[0028] Step 1: adding 5 carbodiimides of 1 mmol to the product of step 3 in claim 1, catalyzing the precipitation of dicyclohexylurea, which was dissolved in water but could not be completely removed in a small amount;

[0029] Step 2: adding a tartaric acid precipitant to the product of step 1, removing dicyclohexylurea through reaction precipitation, filtering the precipitate through a filter, and filtering out the supernatant;

[0030] Step 3: Use a dryer to dry the water to obtain a pure polypeptide product.

[0031] The purity of the prepared polypeptide product is lower than that in Example 1, and the reaction speed and efficiency are lower than that in Example 1, and the cost of the reaction is higher, which is not conducive to a large demand.

Embodiment 3

[0033] A method for preparing the DCC product precipitation of polypeptide synthesis, comprising the following steps:

[0034] Step 1: adding 8 carbodiimides of 1 mmol to the product of step 3 in claim 1, which catalyzes the precipitation of dicyclohexylurea, which cannot be completely removed after being dissolved in water in a small amount;

[0035] Step 2: adding a tartaric acid precipitant to the product of step 1, removing dicyclohexylurea through reaction precipitation, filtering the precipitate through a filter, and filtering out the supernatant;

[0036] Step 3: Use a dryer to dry the water to obtain a pure polypeptide product.

[0037] The purity of the prepared polypeptide product is similar to that of Example 1, and the reaction speed and efficiency are similar to those of Example 1, but the cost of the reaction is very high, which is not conducive to a large demand.

[0038] In summary, when 2 mmol of carbodiimide is added, the precipitated DCC product has the hig...

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Abstract

The invention discloses a preparation method of an amino acid protective agent for polypeptide synthesis and a DCC product precipitate, which comprises the following steps: taking a protective agent X (X is sodium 4-aminobenzene sulfonate), taking 1mmol of the protective agent, dissolving in 10ml of water, fully dissolving, then taking 1mmol of alanine, reacting, combining an X amino group with an alanine carboxyl terminal to form an amido bond, sequentially connecting amino acids such as A, B, C, D, E and the like behind the amino terminal of the alanine, and reacting to obtain the amino acid protective agent for polypeptide synthesis and the DCC product precipitate. According to the present invention, the protective agent is convenient, non-toxic, harmless and easy to precipitate, the X can be combined with the carboxyl terminal of the alanine, can be combined with the precipitating agent to precipitate, and is very practical, the DCC product dicyclohexylurea is precipitated by using another precipitating agent so as to achieve the effect of easily removing the precipitate, such that the use amount of the product is reduced, and the production cost is reduced. A small amount of DCC reaction product dicyclohexylurea is dissolved in water and precipitated by using another precipitator tartaric acid, so that the method is relatively cheap and convenient, the purity extraction efficiency is relatively high, the precision is relatively high, and pure polypeptide is easy to synthesize.

Description

technical field [0001] The present invention relates to the relevant technical field of polypeptide synthesis, in particular to an amino acid protective agent for polypeptide synthesis and a preparation method for DCC product precipitation. Background technique [0002] At present, the molecular structure of the amino acid protective agent used in the polypeptide synthesis technology is complex, and the pollution is relatively large, and it is not easy to precipitate and separate. There is an urgent need for an amino acid protective agent with simple molecules and easy separation to meet market demand. DCC catalysts are added to the synthesis of amino acid chains. A small part of the product dicyclohexylurea is soluble in water and cannot be completely removed. There is a lack of a method for removing the dicycloimine dissolved in water to save costs. This protocol provides a simple and easy technique for the precipitation of amino acid protector primers and dicyclic imines....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K1/06C07K1/02C07K1/30
Inventor 柳春曦柳光旭
Owner 柳春曦
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