Method for preparing chiral amine through transaminase continuous reaction

A transaminase and chemical reaction technology, applied in the field of enzyme catalysis, can solve the problems such as the inability of efficient recycling and regeneration of cofactors and the loss of enzyme amount, and achieve the effect of maintaining the reaction conversion rate unchanged, the reaction conversion rate is basic, and the operation is simple.

Pending Publication Date: 2022-04-12
SHANXI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0005] Aiming at the problems in the prior art that the immobilization of transaminase affects the enzyme activity, the loss of enzyme amount during the circulation of transaminase, and the inability to efficiently recycle and recycle cofactors, the invention provides a fixed-bed continuous flow method based on Pickering emulsion to encapsulate transaminase and cofactor The system is used for the preparation of chiral amine compounds, which effectively avoids the problem of activity loss and loss in the process of enzyme immobilization, and at the same time can realize the synergistic catalysis of transaminase and cofactor in the confined space of emulsion droplets and the recycling of cofactor

Method used

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  • Method for preparing chiral amine through transaminase continuous reaction
  • Method for preparing chiral amine through transaminase continuous reaction
  • Method for preparing chiral amine through transaminase continuous reaction

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Embodiment 1

[0032] A method for preparing chiral amines through transaminase continuous reaction, comprising the steps of:

[0033] Step 1: 1.0 g of SiO with a particle size of 60 nm 2 Nanoparticles were ultrasonically dispersed into 12mL toluene, 4.5mmol octyltrimethoxysilane and 4.5mmol triethylamine were added, stirred and refluxed for 4 hours at 110°C under nitrogen protection, after cooling, the above mixed system was centrifuged to obtain a solid Wash 3-5 times with toluene and dry to obtain interface active nano-SiO 2 (Looks like figure 2 shown);

[0034] Step 2: Add 0.08mL of transaminase ATA-117 solution and 5.0mg of PLP into 3.2mL of 100mM PBS buffer solution, and mix uniformly by magnetic stirring to obtain an aqueous phase system; add 0.32g of interface-active SiO 2 Ultrasonic dispersion in 8mL n-heptane, mixed evenly to obtain an oil phase system; the oil-water two-phase system was mixed, and stirred at a high speed of 5000rpm to form a water-in-oil Pickering emulsion (mo...

Embodiment 2

[0037] A method for preparing chiral amines through transaminase continuous reaction, comprising the steps of:

[0038] Step 1: Add 1.5g of SiO with a particle size of 60nm 2 Nanoparticles were ultrasonically dispersed into 20mL toluene, 4.5mmol dichlorodimethylsilane and 2.25mmol n-hexylamine were added, stirred and refluxed for 6h at 60°C under nitrogen protection, after cooling, the above mixed system was centrifuged, and the obtained solid was used Wash with toluene 3-5 times and dry to obtain interface active nano-SiO 2 ;

[0039] Step 2: Mix 0.72mL transaminase ATA-113 solution, 40mg NADP + Add it to 6.4mL100mM PBS buffer solution, and mix it uniformly by magnetic stirring to obtain an aqueous phase system; add 0.316g of interface-active SiO 2 Ultrasonic dispersion in 16mL of toluene, mixed evenly to obtain an oil phase system; the oil-water two-phase system was mixed, and stirred at a high speed of 5000rpm to form a water-in-oil Pickering emulsion that encapsulated t...

Embodiment 3

[0042] A method for preparing chiral amines through transaminase continuous reaction, comprising the steps of:

[0043] Step 1: 1.2g of SiO with a particle size of 80nm 2 Nanoparticles were ultrasonically dispersed into 20mL toluene, 1.5mmol dichlorodimethylsilane and 4.5mmol n-hexylamine were added, stirred and refluxed for 5h at 60°C under nitrogen protection, after cooling, the above mixed system was centrifuged, and the obtained solid was used Wash with toluene 3-5 times and dry to obtain interface active nano-SiO 2 ;

[0044] Step 2: Add 0.06mL of transaminase ATA-119 solution and 10mg of PLP to 3.2mL of 100mM PBS buffer solution, and mix uniformly by magnetic stirring to obtain an aqueous phase system; add 0.158g of interfacially active SiO 2 Ultrasonic dispersion in 20mL of ethyl acetate, mixed evenly to obtain an oil phase system; the oil-water two-phase system was mixed, and stirred at a high speed of 10000rpm to form a water-in-oil Pickering emulsion encapsulating ...

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Abstract

The invention belongs to the field of enzyme catalysis, and particularly relates to a method for preparing chiral amine through a transaminase continuous reaction. The method comprises the following steps: dispersing transaminase and cofactors in a PBS buffer solution to form a mixed solution; then adding the mixed solution into an oil phase containing interfacial active solid SiO2 nanoparticles, and stirring at a high speed to form a water-in-oil type Pickering emulsion encapsulating transaminase and cofactors; and finally, filling the Pickering emulsion into a fixed bed reactor, and carrying out continuous flow reaction to prepare the chiral amine. The method is simple to operate, the reaction conversion rate is basically kept unchanged after continuous operation for more than 600 hours, the total conversion number of cofactors can reach more than 5000, the transaminase activity is basically kept unchanged, and the cofactors are basically not lost.

Description

technical field [0001] The invention relates to the field of enzyme catalysis, in particular to a method for preparing chiral amine through continuous transaminase reaction. Background technique [0002] Chiral amine compounds widely exist in natural products, drug molecules and multifunctional materials, and are also widely used in organic synthesis as important intermediates, catalysts and chiral auxiliary agents. Therefore, it is necessary to develop efficient methods to synthesize various chiral amines. Sexual amine compounds and their corresponding skeleton structures have important scientific significance and application value. [0003] At present, chemical synthesis is mainly used to synthesize chiral amine drug intermediates in industry. Chemical synthesis requires a variety of dangerous and toxic reagents and heavy metal catalysts, and there are many reaction steps and high reaction temperature, resulting in low yield of chiral amines. It is accompanied by serious ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P13/00C12N9/10
Inventor 杨恒权卫伟樊敏张明
Owner SHANXI UNIV
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