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Method for improving Trametes versicolor laccase output

A technology of Trametes versicolor and laccase, applied in the field of bioengineering, can solve the problems of high-yielding strains, such as difficulty, limited improvement, no advantage in output, etc., to reduce the production cost of laccase, increase the area, and meet the effects of large-scale application.

Active Publication Date: 2015-02-18
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, conventional breeding methods for high-yield strains of laccase are usually heavy workload, low efficiency, and difficult to obtain high-yield strains; Optimization of the amount of inducer can increase the expression level of fungal laccase, but the increase is very limited
Although laccase has been expressed through mature expression systems such as yeast and filamentous fungi, and some high-yielding engineering strains have been obtained, its yield is not superior to that of laccase-producing fungi, and further improvement is needed to meet industrial demands. needs

Method used

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  • Method for improving Trametes versicolor laccase output
  • Method for improving Trametes versicolor laccase output

Examples

Experimental program
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Effect test

Embodiment 1

[0039] Example 1: Activation and pre-cultivation of Trametes versicolor

[0040] Trametes versicolor (Trametes versicolor) was purchased from China Industrial Microorganism Culture Collection Management Center, number CICC14001.

[0041] (1) activation

[0042] The medium for slant activation was prepared by the usual potato medium preparation method, including 200g / L potato, 20g / L glucose, and 15g / L agar powder; Trametes versicolor (CICC14001) was inoculated on the medium, and cultured at 30°C for 7 sky.

[0043] (2) Pre-cultivation

[0044] Prepare the medium for pre-cultivation by using the common potato medium preparation method, including 200g / L potato and 20g / L glucose; inoculate the activated mycelium in the medium, and cultivate it at 25°C and 150r / min 7 days; After homogenizing the culture, inoculate 10% (v / v) inoculum into fresh medium and culture for 5 days to obtain pre-cultured mycelia for laccase fermentation.

[0045] A specific example of Trametes versicolo...

Embodiment 2

[0047] Embodiment 2: Trametes versicolor laccase fermentation

[0048] (1) Under the conditions of temperature 26°C, pH 6.0, stirring rate 400rpm and air flow 0.5vvm, the pre-cultured Trametes versicolor mycelium was inoculated into the laccase fermentation medium with an initial biomass of 2g / L , carry out fermentation culture under this condition;

[0049] (2) Dissolve farnesol in ethanol at a concentration of 200 mM, filter and sterilize with a 0.22 μm filter membrane to prepare a farnesol solution;

[0050] (3) At the 0th hour of the entire culture process, add the farnesol solution in step (2) until the concentration in the fermentation broth reaches 100 μM;

[0051] (4) After 24 hours of cultivation, adjust and maintain the pH at 8.0, adjust the rotation speed to 180rpm, adjust the ventilation to 0.15vvm, and continue to cultivate under these conditions;

[0052] (5) After culturing for 120 hours, collect the fermentation broth, remove the mycelia and measure the lacca...

Embodiment 3

[0055] Embodiment 3: Trametes versicolor laccase fermentation

[0056] (1) Under the conditions of temperature 26°C, pH 4.1, stirring rate 200rpm and ventilation volume 0.2vvm, the pre-cultured Trametes versicolor mycelium was inoculated into the laccase fermentation medium with an initial biomass of 0.5g / L In, carry out fermentation culture under this condition;

[0057] (2) Dissolve farnesol in isopropanol at a concentration of 2 mM, filter and sterilize with a 0.22 μm filter membrane to prepare a farnesol solution;

[0058] (3) At the 36th hour of the entire culture process, add the farnesol solution in step (2) until the concentration in the fermentation broth reaches 10 μM;

[0059] (4) After culturing for 48 hours, adjust and maintain the pH at 6.1, adjust the rotation speed to 80rpm, adjust the ventilation to 0.05vvm, and continue culturing under these conditions;

[0060] (5) After culturing for 168 hours, collect the fermentation broth, remove the mycelium and measu...

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Abstract

The invention discloses a method for improving Trametes versicolor laccase output. The method includes: at the mycelium growth stage of a Trametes versicolor laccase fermentation process, adding farnesol into a fermentation liquor, and further performing fermentation to produce laccase. By applying the method, the laccase output of the Trametes versicolor can be greatly improved. Also, the method is simple in operation, is easy for enlarged production, and is suitable for large-scale laccase fermentation, thereby reducing the laccase production cost, and meeting the requirements of large-scale application.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for increasing the production of Trametes versicolor laccase. Background technique [0002] Oxidoreductase-dominated enzyme-catalyzed oxidation technology has a wide range of industrial applications, including pulp and paper, textile and food industries. Among them, laccase (benzenediol: oxygen oxidoreductase, EC 1.10.3.2) is one of the most promising oxidoreductases, mainly used in organic synthesis, degradation of environmental pollutants and dye decolorization, biological detection And electrochemical analysis, pulp bleaching and wastewater treatment in the paper industry, degradation of fiber raw materials, etc. [0003] Fungal laccase is the main source of laccase. Trametes versicolor, a representative strain of this type of fungus, is considered to be one of the excellent strains for producing laccase, and this fungal laccase has been recognized by some int...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12R1/645
Inventor 刘春朝王锋郭晨
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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