Agrocybe pediades producing laccase, and application of agrocybe pediades producing laccase

A technology of thorn mushroom and laccase, which is applied in the biological field, can solve the problems such as no laccase produced by twig mushroom, and achieves the effects of good application prospect, high enzyme activity and strong temperature stability.

Active Publication Date: 2020-09-22
BIOLOGY INST OF SHANDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there has been no report on the production of laccase by Apes hirata

Method used

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  • Agrocybe pediades producing laccase, and application of agrocybe pediades producing laccase
  • Agrocybe pediades producing laccase, and application of agrocybe pediades producing laccase
  • Agrocybe pediades producing laccase, and application of agrocybe pediades producing laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. Screening and identification of laccase-producing strain Ap-SWS2 of the present invention

[0029] (1) Screening of strains:

[0030] Enrichment culture: In August 2019, soil and weed samples were collected from the southern mountainous area of ​​Jinan, Shandong, and guaiacol was used as a screening agent for screening, and 100 mL of laccase strain screening medium was prepared in a 500 mL Erlenmeyer flask, and the samples were pressed by 1 % ratio was put into the shake flask at 28°C and 180r / min for 96h.

[0031] Preliminary screening: add 100 μL of the enriched sample to an EP tube containing 900 μL sterile normal saline, mix well, then take out 100 μL from it and add it to another EP tube containing 900 μL sterile normal saline, and operate sequentially to dissolve the bacterial liquid were diluted to 10 1 、10 2 、10 3 、10 4 and 10 5 , will be 10 4 and 10 5 The diluted bacterial solution was applied to three parallel laccase screening plates, and p...

Embodiment 2

[0048] Embodiment 2. The cultivation of Ap-SWS2 bacterial strain of the present invention produces laccase

[0049] (1) Medium used

[0050] Seed medium: yeast extract powder 1%, glucose 1.5%, (NH 4 ) 2 SO 4 0.15%, MgSO 4 ·7H 2 O 0.05%, KH 2 PO 4 0.08%, K 2 HPO 4 ·3H 2 O 0.02%, pH 5.5-6.0.

[0051] Fermentation medium 1: MgSO 4 ·7H 2 O 0.05%, KH 2 PO 4 0.08%, K 2 HPO 4 ·3H 2 O 0.02%, (NH 4 ) 2 SO 4 0.15%, CuSO 4 ·5H 2 O 0.00125%, MnSO 4 ·H 2 O 0.0034%, rice straw 4%, pH 5.0.

[0052] Fermentation medium 2: MgSO 4 ·7H 2 O 0.05%, KH 2 PO 4 0.08%, K 2 HPO 4 ·3H 2 O 0.02%, (NH 4 ) 2 SO 4 0.15%, CuSO 4 ·5H 2 O 0.00125%, MnSO 4 ·H 2 O 0.0034%, corn stover 4%, pH 5.0.

[0053] Fermentation Medium 3: MgSO 4 ·7H 2 O 0.05%, KH 2 PO 4 0.08%, K 2 HPO 4 ·3H 2 O 0.02%, (NH 4 ) 2 SO 4 0.15%, CuSO 4 ·5H 2 O 0.00125%, MnSO 4 ·H 2 O 0.0034%, wheat straw 4%, pH 5.0.

[0054] (2) Culture conditions

[0055] Seed liquid culture: Take t...

Embodiment 3

[0059] Example 3. Research on the enzymatic properties of laccase secreted by the Ap-SWS2 bacterial strain of the present invention

[0060] (1) Optimum temperature and temperature stability of laccase

[0061] Enzyme activity was measured at different temperatures (20, 30, 40, 50, 60, 70° C.), the group with the highest activity was set as 100%, and a curve of temperature and relative enzyme activity was obtained. the result shows( image 3 ), the optimal temperature of laccase in the crude enzyme solution is 50°C.

[0062] Place the enzyme solution at different temperatures (0, 10, 20, 30, 40, 50, 60°C) for 1 hour, and measure the enzyme activity at 50°C. The enzyme activity of the untreated enzyme solution is 100%, and the temperature stability and relative enzyme activity are obtained. curve. the result shows( Figure 4 ), the laccase in the crude enzyme solution is relatively stable at temperatures lower than 50°C; when the temperature is higher than 50°C, its stabili...

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Abstract

The invention discloses agrocybe pediades producing laccase, and an application of the agrocybe pediades producing laccase. An agrocybe pediades strain Ap-SWS2 which can secrete laccase is screened for the first time, the strain is preserved in the China General Microbiological Culture Collection Center on 27 June 2020, and the preservation number is CGMCC NO.20209. The fermentation liquid varnishing of the strain is high in enzymatic activity and high in temperature stability, and has better degradation capacity for lignin, and the application of the agrocybe pediades in the fields of crop straw treatment and the like is facilitated.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a laccase-producing strain of Tonus pleurotus and its application. Background technique [0002] my country is rich in straw resources, but because of the concentrated production period and large output of straw, it cannot be used effectively, and most of the discarded straw can only be burned. At present, the problem of waste straw incineration is serious, resulting in a large waste of resources and environmental pollution. Therefore, the high-quality application of straw is very important for straw treatment. Due to the existence of lignin in straw, it is difficult to utilize and degrade it. However, some specific microorganisms can degrade lignin in straw by relying on lignin-degrading enzymes. Biological treatment of straw has become the current research trend of straw bioutilization, but microbial strains that degrade lignin generally have problems such as low acti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/02C12P1/02C02F3/34C12R1/645C02F103/30C02F103/28
CPCC02F3/342C02F2103/28C02F2103/30C12N9/0061C12P1/02C12Y110/03002C12N1/145C12R2001/645
Inventor 王莹楚杰封佳丽
Owner BIOLOGY INST OF SHANDONG ACAD OF SCI
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