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Fermentation production method of fungal laccase and application of laccase

A production method, the technology of fungal laccase, is applied in the field of white rot fungus to cultivate and ferment laccase in a 5L-scale fermentation tank, the preparation of industrial laccase, and the field of laccase preparation, which can solve the problem of unresolved cost and shear force. increase, low enzyme activity, etc., to achieve low production costs, good bleaching effect, and easy follow-up treatment

Inactive Publication Date: 2014-02-12
SHENZHEN LEVEKING BIOLOGY ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when this kind of fermenter is used for the cultivation and fermentation of white rot fungi, there are still some unfavorable factors, mainly in that the growth of white rot fungus cells in the tank is not easy to control, and the disordered expansion of mycelia will increase the viscosity of the fermentation liquid degree, and wound on the mixing impeller, blocking the pipeline, seriously affecting the uniform transmission of materials, heat and oxygen
Although the above problems can be improved by increasing the stirring speed, too high a rotating speed will increase the shear force on the mycelium, inhibit cell reproduction metabolism and secretion of laccase, and even cause cell rupture, which will lead to low laccase production and low enzyme production. live low
[0004] At present, there are few reports on the mature large-scale fermentation process of white rot fungus laccase, and the problem of production cost has not been solved. And the transmission characteristics of oxygen, etc., the production of white rot fungus is not easy to control

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Preparation of potato extract: Weigh 300g of fresh potatoes, peel and slice, boil in 1000mL deionized water for 30min, filter with gauze after cooling, and dilute the filtrate to 1000mL with deionized water.

[0025] PDA medium: 20 g of anhydrous glucose and 20 g of agar were added to each liter of potato extract.

[0026] Seed medium: Add 20g of anhydrous glucose, 1g of peptone, KH to each liter of potato extract 2 PO 4 3g, MgSO 4 ·7H 2 O 3g.

[0027] Fermentation medium: 10 g of pulverized wheat bran, 0.2 mL of special antifoaming agent for fermentation, and 3 mmol of copper sulfate were added to each liter of seed medium.

[0028] 1. Preparation of strain suspension

[0029] The white rot fungus Panus conchatus was transferred from the PDA slant medium into a 300mL Erlenmeyer flask containing 50mL seed medium, and cultured in a shaker at 150rpm at 28°C for 3 days, the culture solution was filtered through sterile gauze, and the mycelium Wash with sterile water...

Embodiment 2

[0043] Preparation of potato extract: Weigh 300g of fresh potatoes, peel and slice, boil in 1000mL deionized water for 30min, filter with gauze after cooling, and dilute the filtrate to 1000mL with deionized water.

[0044] PDA medium: 20 g of anhydrous glucose and 20 g of agar were added to each liter of potato extract.

[0045] Seed medium: add 10g of anhydrous glucose, 10g of peptone, KH 2 PO 4 3g, MgSO 4 ·7H 2 O3g.

[0046] Fermentation medium: Add 20g of crushed peanut shells, 0.2mL of special antifoaming agent for fermentation, and 5mmol of reveratrol to each liter of seed medium.

[0047] 1. Preparation of strain suspension

[0048] The white rot fungus Panus conchatus was transferred from the PDA slant medium into a 300mL Erlenmeyer flask containing 50mL seed medium, and cultured in a shaker at 150rpm at 28°C for 3 days, the culture solution was filtered through sterile gauze, and the mycelium Wash with sterile water and disperse in a high-speed crushing cup for...

Embodiment 3

[0062] Preparation of potato leaching solution: Weigh 200g of fresh potatoes, peel and slice, boil in 1000mL deionized water for 30min, filter with gauze after cooling, and dilute the filtrate to 1000mL with deionized water.

[0063] PDA medium: 20 g of anhydrous glucose and 20 g of agar were added to each liter of potato extract.

[0064] Seed medium: add 50g of anhydrous glucose, 1g of peptone, KH to each liter of potato extract 2 PO 4 3g, MgSO 4 ·7H 2 O3g.

[0065] Fermentation medium: 5 g of crushed soybean meal, 0.2 mL of special antifoaming agent for fermentation, and 1 mmol of guaiacol were added to each liter of seed medium.

[0066] 1. Preparation of strain suspension

[0067] The white rot fungus Panus conchatus was transferred from the PDA slant medium to a 300mL Erlenmeyer flask containing 50mL seed medium, and cultured in a shaking table at 180rpm at 28°C for 2 days, the culture solution was filtered through sterile gauze, and the mycelia Wash with sterile w...

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Abstract

The invention discloses a fermentation production method of fungal laccase and an application of the laccase. The method comprises the following steps: preparing Panus conchatus suspension; inoculating the suspension into a fermentation tank containing 3-5L of fermentation medium and culturing for 10-15 days, thus obtaining fermentation liquor; and finally, filtering or centrifuging the fermentation liquor, thus obtaining a laccase preparation. The method of the invention is simple and convenient in subsequent treatment of the fermentation liquor and low in production cost, and the activity of the Panus conchatus laccase obtained through fermentation is over 200IU / mL. Compared with other domestic and overseas laccase production methods in a fermentation tank scale, the fermentation production method of the fungal laccase, which is disclosed by the invention, has the advantages that the total laccase yield and the laccase activity reach a relatively high level, and the prepared laccase preparation has a good pulp bleaching effect.

Description

technical field [0001] The invention relates to the technical field of fungal fermentation, in particular to a method for preparing industrial laccase, specifically a method for producing laccase by culturing and fermenting white-rot fungi in a 5L-scale fermenter and the lacquer prepared by the method Application of enzyme preparation in pulp bleaching. Background technique [0002] Laccase (Laccase, EC.1.10.3.2) is a copper-containing polyphenol oxidase, which belongs to the same family of ceruloplasmin oxidases as ascorbate oxidase in plants and ceruloplasmin in mammalian plasma. In the past ten years, the function and application of fungal laccase, especially white rot fungus laccase, has attracted more and more researchers' attention. Due to its high redox potential and wide range of substrates, It can catalyze the oxidation of a variety of phenols and aromatic amines. In the presence of some small molecule redox mediators, it can also oxidize non-phenolic structures in...

Claims

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Application Information

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IPC IPC(8): C12N9/02D21C9/10C12R1/645
CPCC12N9/0061C12Y110/03002D21C9/10
Inventor 付时雨傅恺王剑英张清辉王宏
Owner SHENZHEN LEVEKING BIOLOGY ENG
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