Preparation method of immobilized lipase and method for preparing 2-ethylhexanoic acid

A technology for immobilizing lipase and lipase, applied in biochemical equipment and methods, immobilized on/in organic carriers, enzymes, etc., can solve the problem of difficult separation and treatment of homogeneous catalysts, complicated catalyst preparation process, and product selection. Unsatisfactory performance and other problems, to achieve the effect of high catalytic efficiency and specificity, less green environmental protection three wastes, and reduced catalytic resistance

Active Publication Date: 2019-04-12
WANHUA CHEM GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] In summary, in the existing technology of preparing 2-ethylhexanoic acid from 2-ethylhexanal, or its homogeneous catalyst is difficult to separate and handle (such as Mn(OAc) 2 or KOAc or Cu(OAc) 2 etc.), or the catalyst preparation process is cumbersome (such as heteropolyacids and ionic liquids) and highly corrosive, or the process is complicated, the mass transfer effect is poor, and the product selectivity is not ideal.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 2 g of lipase dry powder was prepared into a 1 mg / mL lipase solution with disodium hydrogen phosphate-citric acid buffer solution with a pH of 7.

[0048] 5g of carboxymethyl-β-cyclodextrin and 40mL of polyallylamine solution (molecular weight 5000, concentration 20wt%, viscosity 13mpa·s) were fully stirred and reacted in a constant temperature shaker at 60°C for 8h. Wash with deionized water for 3 times, and then dry in vacuum to obtain the coupling fragment of polyallylamine and carboxymethyl-β-cyclodextrin (PAA-CM-β-CD).

[0049] Put 2g of PAA-CM-β-CD and 100mL of 1mg / mL lipase solution into a constant temperature shaking box at 30°C for culture, shake for 20 hours to obtain a reaction mixture, then centrifuge, and then use disodium hydrogen phosphate of the same pH as above- Wash with citrate buffer several times until no free lipase can be detected in the supernatant. The obtained immobilized lipase was vacuum-dried to obtain the finished product of immobilized li...

Embodiment 2

[0051] 2 g of lipase dry powder was prepared into a 1.5 mg / mL lipase solution with disodium hydrogen phosphate-citric acid buffer solution with a pH of 5.

[0052] 5g of carboxymethyl-β-cyclodextrin and 40mL of polyallylamine solution (molecular weight 8000, concentration 15wt%, viscosity 10mpa·s) were fully stirred and reacted in a constant temperature shaker at 60°C for 8h. Wash with deionized water for 3 times, and then dry in vacuum to obtain the coupling fragment of polyallylamine and carboxymethyl-β-cyclodextrin (PAA-CM-β-CD).

[0053] 2g of PAA-CM-β-CD and 160mL of 1.5mg / mL lipase solution were placed in a constant temperature shaking box at 30°C for culture, and the reaction mixture was obtained after shaking for 20 hours, followed by centrifugation, and then disodium hydrogen phosphate of the same pH as above - Multiple washes with citrate buffer until no lipase is detectable in the supernatant. The obtained immobilized lipase was vacuum-dried to obtain the finished ...

Embodiment 3

[0055] 2 g of lipase dry powder was prepared into a 2 mg / mL lipase solution with disodium hydrogen phosphate-citric acid buffer solution with a pH of 7.

[0056] 5g of carboxymethyl-β-cyclodextrin and 40mL of polyallylamine solution (molecular weight 3000, concentration 20wt%, viscosity 10mpa·s) were fully stirred and reacted in a constant temperature shaker at 60°C for 8h, after the reaction Wash with deionized water for 3 times, and then dry in vacuum to obtain the coupling fragment of polyallylamine and carboxymethyl-β-cyclodextrin (PAA-CM-β-CD).

[0057] 2g of PAA-CM-β-CD and 140mL of 1mg / mL lipase solution were placed in a constant temperature shaking box at 30°C for culture, and the reaction mixture was obtained after shaking for 20 hours, followed by centrifugation, and then disodium hydrogen phosphate of the same pH as above- Wash with citrate buffer several times until no lipase can be detected in the supernatant. The obtained immobilized lipase is vacuum-dried to ob...

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Abstract

The invention relates to a preparation method of immobilized lipase and a method for preparing 2-ethylhexanoic acid. An adopted catalyst is immobilized lipase which is formed in a manner that biological lipase is loaded on carboxymethyl-beta-cyclodextrin through flexible arm poly(allylamine). The defect that enzymic activity can be reduced in a conventional enzyme solidifying technique can be overcome. The immobilized enzyme is efficient, simple and convenient to prepare, is environmental-friendly, and can be cycled, regenerated and used. The immobilized lipase is used for preparing the 2-ethylhexanoic acid through oxidation of 2-ethylhexaldehydes, and the defects of poor selectivity, difficult catalyst separation, serious three wastes and the like in a conventional 2-ethylhexanoic acid preparation technique can be overcome.

Description

technical field [0001] The invention relates to a preparation method of an oxidation reaction catalyst, in particular to a preparation method of an immobilized lipase catalyst used for oxidizing 2-ethylhexanal to 2-ethylhexanoic acid. Background technique [0002] 2-Ethylhexanoic Acid (2-EHA), also known as isooctanoic acid in Chinese, is an oily liquid slightly soluble in cold water and ethanol, soluble in hot water and ether. It is characterized by high purity and light color. It can react with a variety of metal compounds to form 2-ethylhexanoic acid metal salts. Its main uses are as follows: drier for coatings, auxiliary agent for lubricating oil, plasticizer for plastics, stabilizer for PVC, salt-forming agent for medicine, and auxiliary agent for reaction catalysts . [0003] At present, my country has become the world's second largest paint producer next to the United States, especially the rapid development of high-grade paint. In addition, my country's synthetic r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/08C12P7/40
CPCC12N9/20C12N11/08C12P7/40C12Y301/01003
Inventor 王泽圣曹善健丛鑫何光文刘超董科王鹏崔乾余炎冰
Owner WANHUA CHEM GRP CO LTD
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