Mesophilic laccase gene, mesophilic laccase and application thereof

A kind of laccase and gene technology, applied in the application field of mesophilic laccase ZGL3, textile industry and environmental protection field

Inactive Publication Date: 2014-02-26
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few research reports on the degradation and decolorization of bacterial laccases in dyes. By studying the decolorization effects of bacterial laccases on different synthetic dyes, the industrial application of bacterial laccases can be further promoted.

Method used

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  • Mesophilic laccase gene, mesophilic laccase and application thereof
  • Mesophilic laccase gene, mesophilic laccase and application thereof
  • Mesophilic laccase gene, mesophilic laccase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: expression and preparation of mesophilic laccase ZGL3 gene cloning and mesophilic laccase ZGL3, comprising the following steps:

[0026] (1) Cultivation of mesophilic bacteria ZW2531-1 and extraction of chromosomal DNA

[0027] The medium composition of each 100ml mesophilic bacteria ZW2531-1 is as follows: yeast powder 0.4g, peptone 0.08g, NaCl 0.3g, medium pH=7.6; culture conditions are: 50°C shaker at 200rpm constant temperature culture About 8 hours; ZW2531-1 stem cells were added to 1 mL of the above-mentioned medium and cultured for 8 hours, then transferred to 100 mL of fresh above-mentioned medium for expansion for 8 hours, centrifuged at 8000 rpm for 5 minutes to collect the cultured mesophilic cells, and stored at -20°C for future use.

[0028]Take 0.2g of the mesophilic bacteria collected above, wash the bacteria once with 400μL, 25mmol / L Tris-HCl buffer (pH=8.0, containing 50mmol / L glucose, 10mmol / L EDTA), centrifuge and discard the supernatant...

Embodiment 2

[0047] Example 2: Characterization of the mesophilic laccase ZGL3

[0048] (1) Optimum reaction pH and pH stability

[0049] pH value is an important factor affecting the catalytic activity of enzymes. Usually the enzyme shows the maximum catalytic activity at a certain pH value, and the farther it deviates from this pH value, the lower the catalytic activity of the enzyme. The maximum pH range detected by the catalytic activity of the mesophilic laccase ZGL3 of the present invention is 2.2~ 10.0.

[0050] Using ABTS and DMP as substrates, measure the activity of mesophilic laccase ZGL3 in buffers with different pH values ​​(2.2.0-10.0), and calculate the relative enzyme activity under other pH conditions with the highest enzyme activity as 100%. . The relative activity of the enzyme is plotted against the pH value ( Figure 5 A), the results show that the optimum pH values ​​of ABTS and DMP catalyzed by mesophilic laccase ZGL3 are 4.0 and 7.5, respectively.

[0051] After ...

Embodiment 3

[0056] Example 3: Efficacy Analysis of Mesophilic Laccase ZGL3 Catalytic Dye Decolorization

[0057] Using ABTS and syringaldehyde as mediator substrates respectively, the mesophilic laccase ZGL3 was analyzed for five dyes reactive blue 19 (Remazol Brilliant Blue R, RBBR), Congo red (Congo red), indigo carmine (indigo carmine), formazan The decolorization efficiency of different concentrations of base red (MethyI red) and crystal violet (Crystal violet) (100mg / L, 10mg / L, 40mg / L, 50mg / L, 5mg / L), the decolorization rate is calculated by the following formula: decolorization Rate% = (initial absorbance value - absorbance value after catalysis) / initial absorbance value × 100%.

[0058] Table 1. Efficacy analysis of dye decolorization catalyzed by mesophilic laccase ZGL3

[0059]

[0060] The results are shown in Table 1, Figure 7 shown. In the absence of mediator substrates, mesophilic laccase ZGL3 can effectively decolorize RBBR, Congo red and indigo carmine, but is less...

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Abstract

Mesophilic laccase gene, mesophilic laccase and application thereof belong to the field of biotechnology. In particular, the invention relates to a mesophilic laccase gene from mesophilic bacteria ZW2531-1, mesophilic laccase expressed by the gene ZGL3 and application of the mesophilic laccase to the fields of textile industry and environmental protection. More specifically, the invention provides a mesophilic laccase gene derived from a thermophilic bacterium and mesophilic laccase expressed by the gene. The invention also relates to the effective application of the thermophilic laccase to industrial dye degradation, and further relates to the wide application of the mesophilic laccase to the fields of textile industry and environment protection. The mesophilic laccase provided by the invention can safely and effectively realize degradation and decolorization of anthraquinone, azo and triphenylmethane dyes, has broad application prospects, easiness for realizing industrialization preparation and high practical value.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a mesophilic laccase gene derived from a mesophilic bacterium ZW2531-1, a mesophilic laccase ZGL3 expressed by the gene, and the use of the mesophilic laccase in the fields of textile industry and environmental protection, etc. in the application. Background technique [0002] Laccase (EC1.10.3.2) is a copper-containing polyphenol oxidase, which can use copper ions in the active center to catalyze the oxidation of aromatic compounds with various structures, and at the same time reduce molecular oxygen to water. Due to the wide range of substrates and high catalytic efficiency of laccase, it is widely used in paper industry (biological pulping and bleaching), textile industry (decolorization of synthetic dyes), textile fiber softening, food industry (food flavor improvement), and feed nutrition. Improvement, new drug development, soil bioremediation, biosensor man...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C02F3/00C02F103/34C02F101/38
CPCC12N9/0061C12Y110/03002
Inventor 张应玖高键关可兴
Owner JILIN UNIV
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