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Method for improving laccase activity for organophosphorus pesticide degradation

An organophosphorus pesticide and laccase technology, which is applied in the field of pesticide degradation to achieve the effects of reducing production cost, increasing yield and optimizing process conditions

Inactive Publication Date: 2020-01-10
孙芳
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Trametes vermilion ( Trametes cinnabarina (Jacq.) Fr.), aliased as Trametes erythrosum and Thrombus, belonging to the genus Trametes in the family Polyporaceae. Studies have shown that Trametes vermilion is a white-rot fungus with a strong ability to produce laccase, but the use of Trametes vermilion There is no report on the production of laccase by bacteria fermentation

Method used

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  • Method for improving laccase activity for organophosphorus pesticide degradation
  • Method for improving laccase activity for organophosphorus pesticide degradation
  • Method for improving laccase activity for organophosphorus pesticide degradation

Examples

Experimental program
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Embodiment 1

[0032] A method for improving the activity of organophosphorus pesticide degradation laccase, comprising the following steps:

[0033] 1) Activation of strains: Trametes vermilion ( Trametes cinnabarina (Jacq.) Fr.) strains were purchased from Ningbo Mingzhou Biotechnology Co., Ltd. Under sterile conditions, the preserved strains were inoculated on a PDA medium plate, and cultured at a constant temperature of 28°C until the mycelium covered the surface of the plate.

[0034] 2) Cultivation of seed liquid: Punch out 5 pieces of fungus cakes from the activated plate with a 5mm hole puncher, inoculate them into the seed medium, fill the shake flask with a liquid volume of 100mL / 250mL, culture at 26°C and 150r / min for 2 days, and prepare into a first-grade seed solution; re-inoculate the first-grade seed solution into fresh seed medium with an inoculum volume of 6%, and the liquid volume in the shaker flask is 300mL / 1000mL, and culture at a constant temperature of 26°C and 150r / ...

Embodiment 2

[0043] Example 2 Effects of fermentation medium components on laccase production by Trametes vermilion

[0044] 2.1 Effects of different carbon sources on laccase production by Trametes vermilion

[0045] Using 30g / L glucose, fructose, cornstarch, maltodextrin and molasses as the fermentation medium to replace the carbon source, keeping the other components of the medium unchanged, carrying out fermentation and enzyme production according to the method in Example 1, and after 7 days, respectively Prepare crude enzyme solution to measure laccase activity, the experimental results are as follows figure 1 shown.

[0046] from figure 1 It can be seen that when glucose is used as the carbon source, the laccase activity is the highest, reaching 122.45U / mL, followed by maltodextrin as the carbon source, the enzyme activity is 110.32U / mL, and when molasses is used as the carbon source, the enzyme activity is the lowest, which is 68.59U / mL . As one of the most important component...

Embodiment 3

[0056] The influence of embodiment 3 feeding on the production of laccase by Trametes vermilion

[0057] On the basis of Example 1, a fed-batch fermentation experiment was carried out: the secondary seed liquid was homogenized for 30 seconds with a homogenizer under aseptic conditions, and then it was inserted into the sterilized fermentation medium by the flame inoculation method, and inoculated The volume is 8%, the liquid volume in the fermentation tank is 60% (v / v), the fermentation temperature is 26°C, the ventilation volume is 0.75vvm, and the stirring speed is 220r / min; the fermentation process adopts feeding 1mol / L ammonia water and 1mol / L hydrochloric acid to adjust the pH of the fermentation broth, and maintain the pH value at 5.0~5.5; after 3 days of fermentation, add maltodextrin according to the feeding amount of 10g / L, 20g / L, and 30g / L, and feed every 24h 1 time, 3 consecutive additions, with no addition of maltodextrin as a control, crude enzyme solutions were ...

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Abstract

The invention relates to the technical field of pesticide degradation and particularly relates to a method for improving the laccase activity for organophosphorus pesticide degradation. The method comprises the following steps: (1) activating a culture; (2) culturing a seed solution; (3) performing fermentation enzyme-production culture; and (4) preparing a crude enzyme solution. According to a fermentation culture medium, maltodextrin is used as a carbon source, corn protein and peptone are used as a complex nitrogen source, and substances capable of promoting laccase synthesis and secretionof trametes cinnabarina, such as tyrosine, salicylic acid, diethylaminoethyl hexanoate, catechin, veratryl alcohol, a fullerene aqueous solution, spermidine and tween-80, are added into the culture medium, feed-batch and pH regulation are performed in the fermenting process, and an optimized production process for producing laccase by trametes cinnabarina liquid fermentation is established, so that the aims of improving the activity and yield of laccase and reducing the production cost of laccase can be achieved, and foundation is laid for industrial production of laccase and application in the field of pesticide degradation in the future.

Description

technical field [0001] The invention relates to the technical field of pesticide degradation, in particular to a method for improving the activity of organophosphorus pesticide degradation laccase. Background technique [0002] The types of pesticides can be divided into organic chlorine, organic phosphorus, organic nitrogen, organic sulfur, arsenic preparations, pyrethroids, etc. according to their chemical composition. Since the 1960s, many countries have banned or restricted the use of organochlorine pesticides, while organophosphorus pesticides have gradually developed into the most widely used pesticide category due to their variety, high efficacy, and broad spectrum of control. At present, there are more than 150 types of organophosphorus pesticides in the world, and there are more than 20 types of organophosphorus pesticides produced in my country, with an annual output of more than 100,000 tons, accounting for more than 80% of my country's total pesticide output. Or...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N1/14A62D3/02C12R1/645A62D101/04A62D101/26A62D101/28
CPCC12N9/0061C12Y110/03002C12N1/14A62D3/02A62D2101/04A62D2101/26A62D2101/28
Inventor 孙芳
Owner 孙芳
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