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Method for preparing immobilized lipase by using porous nano-gold as carrier, and application of immobilized lipase

A technology of porous nano-gold and fat immobilization, applied in biochemical equipment and methods, immobilized on or in inorganic carriers, enzymes, etc., can solve the problems of poor stability of free enzymes, difficult separation of products, and inability to recycle, etc. High reuse efficiency and good stability

Inactive Publication Date: 2018-11-23
TAIYUAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantages of free enzymes such as poor stability, difficult separation from products, and inability to recycle hinder their further application in industrial production.

Method used

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  • Method for preparing immobilized lipase by using porous nano-gold as carrier, and application of immobilized lipase
  • Method for preparing immobilized lipase by using porous nano-gold as carrier, and application of immobilized lipase

Examples

Experimental program
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Effect test

Embodiment 1

[0033] A preparation method for immobilizing lipase with porous nano-gold as a carrier, comprising the following steps:

[0034] 1. Preparation of porous gold nanomaterials

[0035] (1) Cut a gold-silver alloy material with a gold-silver atomic ratio of 48:52 into a 1cm×1cm square sample, weighing 5 mg. The sample was placed in concentrated nitric acid with a mass concentration of 68%, and corroded at room temperature for 24 hours. Wash the sample five times with ultrapure water until neutral, and dry the sample in a vacuum freeze dryer.

[0036] (2) Put the above sample into the annealing furnace, set the temperature at 200°C, and start timing for two hours after the temperature rises to the preset temperature. The annealing furnace was closed, and the porous nano-gold (NPG) material was obtained after natural cooling.

[0037] Under the above process conditions, a nanoporous material with a three-dimensional continuous structure can be prepared. Such as figure 1 shown. ...

Embodiment 2

[0049] A preparation method for immobilizing lipase with porous nano-gold as a carrier, comprising the following steps:

[0050] 1. Preparation of porous gold nanomaterials

[0051] (1) Cut a gold-silver alloy material with a gold-silver atomic ratio of 48:52 into a 1cm×1cm square sample, weighing 5mg. The sample was placed in concentrated nitric acid with a mass concentration of 70%, and corroded at room temperature for 24 hours. Wash the sample 6 times with ultrapure water until neutral, and put it into a vacuum freeze dryer to dry the sample.

[0052] (2) Put the above sample into the annealing furnace, set the temperature at 250°C, and start timing for two hours after the temperature rises to the preset temperature. The annealing furnace was closed, and the porous nano-gold (NPG) material was obtained after natural cooling.

[0053] 2. Covalently linked lipase with gold nanomaterials

[0054] (1) Dissolve α-lipoic acid solid in absolute ethanol solution to prepare 10ml...

Embodiment 3

[0061] A preparation method for immobilizing lipase with porous nano-gold as a carrier, comprising the following steps:

[0062] 1. Preparation of porous gold nanomaterials

[0063] (1) Cut a gold-silver alloy material with a gold-silver atomic ratio of 50:50 into a 1cm×1cm square sample, weighing 5mg. The sample was placed in concentrated nitric acid with a mass concentration of 75%, and corroded at room temperature for 16 hours. Wash the sample five times with ultrapure water until neutral, and dry the sample in a vacuum freeze dryer.

[0064] (2) Put the above sample into the annealing furnace, set the temperature at 300°C, and start timing for 1.5 hours after the temperature rises to the preset temperature. The annealing furnace was closed, and the porous nano-gold (NPG) material was obtained after natural cooling.

[0065] Under the above process conditions, a nanoporous material with a three-dimensional continuous structure can be prepared. Such as figure 1 shown. ...

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Abstract

The invention discloses a method for preparing immobilized lipase by using porous nano-gold as a carrier, and application of the immobilized lipase. According to the method, the porous nano-gold material is prepared by using a dealloying method; the method comprises the steps of firstly, modifying by using alpha-lipoic acid so as to form a carboxylated nano-gold material; modifying the carboxylated nano-gold material by using a coupling agent so as to form a nano-gold compound terminated by N-hydroxysulfosuccinimide sodium salt; immobilizing porcine pancreatic lipase onto the nano-gold compound. The immobilized lipase prepared by the method has good catalytic activity, pH stability and thermal stability; compared with resolvase, the immobilized lipase has obviously improved storage stability; furthermore, after being subjected to simple treatment, the carrier can be recycled, so that the cost is lowered.

Description

technical field [0001] The invention relates to a preparation method and application of immobilizing lipase with porous nano-gold as a carrier, and belongs to the technical field of biological material preparation. Background technique [0002] Lipase has a wide range of applications in food processing, medicine, sensors and other fields due to its high selectivity in catalytic reactions such as oil hydrolysis, transesterification, and lipid reaction. However, the disadvantages of free enzymes such as poor stability, difficult separation from products, and inability to recycle hinder their further application in industrial production. Immobilizing the free enzyme on a special carrier by physical or chemical methods can improve its basic properties such as thermal stability and storage stability while maintaining its natural activity. In addition, the combination with a special functional carrier can also meet It needs special functionalization in biology and other fields. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14
CPCC12N9/20C12N11/14C12Y301/01003
Inventor 杨晓宁黄晓波刘晓晔王永康张孟唐宾
Owner TAIYUAN UNIV OF TECH
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