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Method for improving yield of laccase of Trametes versicolor

A technology of Trametes versicolor and laccase, which is applied in the field of bioengineering, can solve the problems of less research on biological effects, unclear theoretical mechanism of biological effects, etc., and achieves the effects of increasing the yield of laccase, increasing the area, and being easy to enlarge the production.

Active Publication Date: 2018-07-17
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, few researchers have studied the biological effects of ultrasound, and the theoretical mechanism of the biological effects is not clear

Method used

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  • Method for improving yield of laccase of Trametes versicolor

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Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1: Trametes versicolor laccase fermentation

[0032] (1) Inoculate the pre-cultivated Trametes versicolor mycelium into the laccase fermentation medium with an initial biomass of 10% (v / v) at a temperature of 26°C and 150r / min. fermentation culture;

[0033] (2) During the 6th to 48h of the fermentation process, the bacteria in the triangular flask were ultrasonically treated, the frequency was 45kHz, the ultrasonic power was 120w, and the ultrasonic time was 10s and 10s, 5min, 2 times; 60-120 hours, ultrasonic treatment was performed on the bacteria, the frequency was 20kHz, the power was 150w, and the number of times was 3 times. The rest of the conditions were unchanged, and the cultivation was continued after ultrasonication.

[0034] (3) From the first day to the third day of the fermentation process, adjust the pH to 4.5.

[0035] (4) After 7 days of fermentation and cultivation, the fermentation liquid was collected, and the production of laccase was...

Embodiment 2

[0037] Embodiment 2: Trametes versicolor laccase fermentation

[0038] (1) Inoculate the pre-cultivated Trametes versicolor mycelium into the laccase fermentation medium with an initial biomass of 10% (v / v) at a temperature of 26°C and 150r / min. fermentation culture;

[0039] (2) During the 6th to 48h of the fermentation process, the bacteria in the triangular flask were ultrasonically treated, the frequency was 50kHz, the power was 90w, the time was 10s and 10s, 10min in total, and the ultrasonic frequency was 1 time; ~ 120h, ultrasonic treatment was carried out on the bacteria, the frequency was 28kHz, the power was 120w, the number of ultrasonic waves was 2 times and other conditions remained unchanged, and the cultivation was continued after ultrasonication.

[0040] (3) From the first day to the third day of the fermentation process, adjust the pH to 4.5.

[0041] (4) After 7 days of fermentation and cultivation, the fermentation liquid was collected, and the production...

Embodiment 3

[0044] Embodiment 3: Trametes versicolor laccase fermentation

[0045] (1) Inoculate the pre-cultivated Trametes versicolor mycelium into the laccase fermentation medium with an initial biomass of 10% (v / v) at a temperature of 26°C and 150r / min. fermentation culture;

[0046] (2) During the 6th to 48h of the fermentation process, the bacteria in the triangular flask were ultrasonically treated, the frequency was 60kHz, the power was 60w, the time was 20s and 20s, 20min in total, and the number of ultrasounds was 1 time; ~ 120h, ultrasonic treatment was carried out on the bacteria, the frequency was 28kHz, the power was 120w, the number of ultrasonic waves was 2 times and other conditions remained unchanged, and the cultivation was continued after ultrasonication.

[0047] (3) From the first day to the third day of the fermentation process, adjust the pH to 4.5.

[0048] (4) After 7 days of fermentation and cultivation, the fermentation liquid was collected, and the productio...

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Abstract

The invention discloses a method for improving the laccase yield of Trametes versicolor. In the Trametes versicolor laccase fermentation process, fungi are subjected to ultrasonic treatment and continuous fermentation to generate laccase. By directly conducting ultrasonic treatment on the fungi in a shake flask in the fermentation process, operations are simple, and the contamination probability is reduced. In the mycelia growth stage, fungus mycelia have hyperbranched mycelium morphology and bulbous tops through high-intensity ultrasound so as to increase the area of secretion sites, and themembrane permeability is improved so as to accelerate the secretion of laccase. In the laccase generation stage, reactive oxygen type substances can be induced to be generated through low-intensity ultrasound so as to stimulate the expression of a laccase gene, and therefore the laccase yield of Trametes versicolor is effectively improved. It is shown through experiments that the laccase yield canbe improved by three times or above and 5-6 times at most by means of the method. The method is easy to operate, beneficial for scale-up production and suitable for large-scale laccase fermentation,thereby reducing laccase production cot and meeting the large-scale application requirement.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for increasing the yield of Trametes versicolor laccase. Background technique [0002] Laccase (laccase, EC1.10.3.2) is a copper-containing polyphenol oxidase that catalyzes a variety of phenolic compounds by obtaining oxygen to generate corresponding benzoquinones and water. In nature, laccase mainly exists in plants, fungi, and a few insects and bacteria. Trametes versicolor, as one of the recognized excellent strains for producing laccase, is the main "producer" of laccase. In recent years, laccases have received more and more attention due to their industrial applications, such as wastewater treatment, bioremediation, pulp bleaching, establishment of biosensors, and food processing. However, the low level of laccase fermentation expression becomes one of the reasons for its high production and application costs. Therefore, increasing the yield of laccase and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/02C12N1/14C12R1/645
CPCC12N1/14C12N9/0061C12Y110/03002
Inventor 王锋徐玲马海乐孙柯
Owner JIANGSU UNIV
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