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Immobilized spore laccase as well as preparation method and application thereof

A technology for spore laccase and spore, which is applied to immobilized spore laccase and its preparation and application fields, can solve the problems of difficult porous material adsorption, inextensibility, large weight, etc., and achieves strong decolorization ability, strong stability, and easy operation. Effect

Active Publication Date: 2015-07-01
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, according to the characteristics of spores and immobilization methods and the existing reports on spore immobilization, the immobilization of spores is more difficult than the immobilization of general enzymes or bacteria.
The immobilization of spores is different from the immobilization of general enzymes. It is bulky, heavy, and inextensible, so it is difficult to use porous materials to adsorb it. Generally, the pore size of porous materials is smaller than that of spores, and the hydrogen bonds formed are not enough to stabilize large spores. It is also difficult to fix it by embedding, and it has a very serious diffusion limitation phenomenon, and it is also difficult to fix it by covalent bonding and cross-linking. The less contact surface between the spore and the carrier is difficult to provide enough The force of the force makes the spores fixed on the carrier

Method used

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  • Immobilized spore laccase as well as preparation method and application thereof
  • Immobilized spore laccase as well as preparation method and application thereof
  • Immobilized spore laccase as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0048] The immobilization method 1 of embodiment 1 spore laccase

[0049] Add 5 column volumes of deionized water to swell the DEAE-cellulose, stir, and let stand overnight. Suction filter the swollen DEAE-cellulose, then treat with 0.5M HCl for 1 hour, wash with water until neutral, filter, then treat with alkali for 1 hour, wash with water until neutral, filter and collect. Mix the activated DEAE-cellulose with the prepared spores at a ratio of 50mg wet weight spores: 1g dry weight DEAE-cellulose, adsorb at 37°C and 180r / min for 10h, then filter, collect and immobilize Spore laccase and filtrate and detection of enzyme activity. The final immobilization rate of immobilized spore laccase was 99.23%. In the detection of the influence of the amount of spores, the relative enzymatic activity of the immobilized spore laccase obtained under the experimental conditions reached 98.22% of the maximum value. In the detection of the influence of adsorption time, the relative enzymat...

Embodiment 2

[0050] The immobilization method 2 of embodiment 2 spore laccase

[0051] The adsorption pH was 8, and the other steps were the same as in Example 1. The final immobilization rate of immobilized spore laccase was 90.41%.

Embodiment 3

[0052] The immobilization method 3 of embodiment 3 spore laccase

[0053] The adsorption pH was 5, other steps were the same as in Example 1, and the immobilization rate of the final immobilized spore laccase was 80.73%.

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Abstract

The invention discloses immobilized spore laccase as well as a preparation method and an application thereof, and belongs to the immobilized technical field and the wastewater treatment field. The immobilized spore laccase is formed by immobilizing spore laccase produced by a bacillus bacterial strain on DEAE(diethyl-aminoethanol)-cellulose through an ionic bond and a hydrogen bond, and has the extensive pH adaptive capacity and the heat resistnace of the spore laccase; in a solution containing an organic solvent, metal ions or an inhibitor, the immobilized spore laccase has stability which is stronger than that of free spore laccase. The preparation method of the immobilized spore laccase mainly comprises four steps of spore preparation, carrier pre-treatment, adsorption and collection, is simple and convenient to operate, and high in immobilization rate. The immobilized spore laccase provided by the invention has very strong dye discoloring capacity and has a highest discoloring rate up to about 95% while being applied to spinning dyes of methyl green, methyl red, cid red 1 and crystal violet. The carrier also can selectively adsorb the dyes to form complementary action with laccase.

Description

technical field [0001] The invention relates to an immobilized spore laccase and its preparation and application, belonging to the technical field of immobilization and the field of wastewater treatment. Background technique [0002] Laccase is a copper-containing polyphenol oxidase widely found in fungi, bacteria, plants and insects. Laccase can oxidize many phenolic and nonphenolic substances, including monophenols, diphenols, polyphenols, aminophenols, methoxyphenols, aromatic amines, and ascorbates. Laccase can reduce oxygen to water while oxidizing the substrate, and it will not produce harmful substances, so it is a very environmentally friendly biocatalyst. Laccase has good application prospects in paper industry, wastewater treatment, textile industry, food processing, biosensor and biodiagnosis due to its broad substrate spectrum and strong oxidizing ability. [0003] In recent years, scholars have discovered that bacterial laccases are extremely stable in alkalin...

Claims

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Application Information

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IPC IPC(8): C12N11/12C02F3/34C02F103/30
Inventor 管政兵周稳廖祥儒蔡宇杰
Owner JIANGNAN UNIV
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