Magnetic immobilized spore laccase and preparing method and application thereof

A spore laccase and magnetic technology, applied in chemical instruments and methods, immobilized on or in inorganic carriers, and textile industry wastewater treatment, etc. The effect of preparation cost and high decolorization efficiency

Active Publication Date: 2015-11-04
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In recent years, the research on spore laccase is still in the description of biochemical characteristics and the application of simple dye decolorization, and there are very few reports on the immobilization of spore laccase

Method used

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  • Magnetic immobilized spore laccase and preparing method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] The production of embodiment 1 spore laccase

[0036] Cultivate the Bacillus pumilus W3 strain on the spore-forming medium at 37°C for 2 days, centrifuge the above culture at a centrifugal speed of 8000rpm for 10 minutes, collect the spores, then wash the spores once, and add lysozyme to the washed spores Keep the final concentration at 1mg / ml, incubate at 37°C for 2h, treat the culture with lysozyme, wash with 1M NaCl and 1M KCl successively, and collect spores by centrifugation at 8000rpm for 10min; wet weight the spores, and use Resuspend the spores in deionized water at a concentration of 100 mg / ml.

Embodiment 2

[0037] Example 2 Immobilization Scheme 1

[0038] Weigh 0.1g Fe 3 o 4 Magnetic nanoparticles (particle size is 20nm), added to 5ml 0.02M Na 2 HPO 4 -NaH 2 PO 4 Buffer (pH 7, containing 0.1M NaCl), add 0.5ml carbodiimide hydrochloride (4mg / ml), sonicate for 10min, then add 0.2ml spores (100mg / ml, wet weight), sonicate for 30min , and then placed at 200rpm, rotating and shaking at 37°C for 60min; the immobilized mixture was separated by magnetic adsorption, washed with buffer three times, and the supernatant and immobilized spores were collected and stored at 4°C. The recovery rate of enzyme activity was measured to be 92.97%.

Embodiment 3

[0039] Example 3 Immobilization scheme 2

[0040] Weigh 0.1g Fe 3 o 4 Magnetic nanoparticles (particle size is 20nm), added to 10ml 0.02M Na 2 HPO 4 -NaH 2 PO 4 Buffer (pH 7, containing 0.1M NaCl), add 0.5ml carbodiimide hydrochloride (4mg / ml), sonicate for 10min, then add 0.2ml spores (100mg / ml, wet weight), sonicate for 30min , and then placed at 200rpm, rotating and shaking at 37°C for 60min; the immobilized mixture was separated by magnetic adsorption, washed with buffer three times, and the supernatant and immobilized spores were collected and stored at 4°C. The recovery rate of enzyme activity was measured to be 90.12%.

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Abstract

The invention discloses magnetic immobilized spore laccase and preparing method and application thereof and belongs to the field of immobilization technologies and waste water processing. According to the magnetic immobilized spore laccase and the preparing method and application thereof, Fe3O4 magnetic nanoparticles serve as a carrier of the immobilized spore laccase, and the magnetic nanoparticles are directly fixed to the surfaces of spores through a bifunctional crosslinking agent; the magnetic immobilized spore laccase can be quickly separated through an external magnetic field and recycled. According to the magnetic spore laccase and preparing method, surface modification does not need to be conducted on the magnetic nanoparticles, the process just includes spore preparation, immobilization processing and collection, the operation requirement of the preparing process is greatly lowered, and the preparing cost of the immobilized spore laccase is further greatly saved. The magnetic immobilized spore laccase can be directly used for decolorizing of dyes, and high decolorizing efficiency is kept.

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 distributed in fungi, bacteria, plants and insects. Laccase can oxidize phenolic substances, and with the help of mediators, it can also oxidize non-phenolic substances. It has been studied and applied in dye decolorization, beverage treatment, pulp bleaching and biosensor development. Spore laccase refers to the laccase fixed on the spore coat. It is the most representative bacterial laccase. It uses spores as the research unit and has the characteristics of alkali resistance and high temperature resistance. It has obvious advantages in the application of textile dye decolorization. [0003] In recent years, the research on spore laccase is still in the description of biochemical charact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C02F3/34C02F103/30
Inventor 管政兵周稳廖祥儒蔡宇杰伊海希
Owner JIANGNAN UNIV
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