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Method for producing laccase by composite fermentation of white-rot fungus

A technology of composite fermentation and white rot fungus, which is applied in the field of bioengineering and microbial fermentation, can solve the problems such as the effect of enzyme production that has not yet appeared, and achieve the effects of high efficiency and stability of products, high efficiency of laccase production, and easy control

Inactive Publication Date: 2012-09-19
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few domestic reports on the industrial production of laccase, and the production of laccase is still in the stage of theoretical exploration and experimentation
Moreover, the current fermentation and cultivation methods of laccase are mainly limited to the liquid or solid fermentation culture of single bacteria, and the research is mainly aimed at optimizing the culture medium and fermentation conditions of liquid or solid fermentation culture of single bacteria, and there has not been any particularly prominent one that can be industrialized. Production Enzyme production effect

Method used

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  • Method for producing laccase by composite fermentation of white-rot fungus
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  • Method for producing laccase by composite fermentation of white-rot fungus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0021] Example 1-3 Comparison of laccase enzyme activity produced by different combinations of bacteria using corn cob powder raw materials

[0022] (1) Preparation of culture medium: take glucose 1.0g, peptone 0.03g, KH 2 PO 4 0.060g, ZnSO 4 0.0001g, K 2 HPO 4 0.04g, FeSO 4 0.00005g, MnSO 4 0.005g, MgSO 4 0.050g, add 1000mL distilled water, mix well, heat to dissolve, adjust the pH to 6.0, and obtain the basic liquid medium; divide the basic liquid medium into 250mL Erlenmeyer flasks, and the basic liquid medium contained in each Erlenmeyer flask 30 mL, each of which was added 3.0 g of corncob powder crushed to 60 mesh, and stirred evenly to obtain a culture solution.

[0023] (2) Activation of bacterial strains: Inoculate Pleurotus radiatus (PR), Pyriophthora cerevisiae (DS) and Cerexoides cerevisiae (CS) on the potato dextrose agar plate medium of the same thickness that has been sterilized respectively , activated and cultured at 28°C for 7 days.

[0024] (3) Inoc...

Embodiment 4-7

[0031] Example 4-7 Comparing the Enzyme Activity of Laccase Produced by Different Raw Materials by Combination Bacteria Phytophthora radiata and Pyreophthora filatus

[0032] (1) Preparation of culture medium: take glucose 1.0g, peptone 0.03g, KH 2 PO 4 0.060g, ZnSO 4 0.0001g, K 2 HPO 4 0.04g, FeSO 4 0.00005g, MnSO 4 0.005g, MgSO 4 0.050g, add 1000mL distilled water, mix well, heat to dissolve, adjust the pH to 6.0, and obtain the basic liquid medium; divide the basic liquid medium into 250mL Erlenmeyer flasks, and the basic liquid medium contained in each Erlenmeyer flask 30 mL, add 3.0 g of corn stalk powder crushed to 60 mesh, or 3.0 g of corn stalk powder crushed to 60 mesh, or 3.0 g of wheat stalk powder crushed to 60 mesh, or 3.0 g of straw powder crushed to 60 mesh, and stir The culture solution containing different crop stalk powder raw materials can be obtained evenly.

[0033] (2) Activation of the strains: Inoculate Pleurotus radiata (PR) and Pyriophthora fi...

Embodiment 8-9

[0039] Example 8-9 Effects of Different Glucose Concentrations in the Culture Solution on the Laccase Production of the Combination Bacteria of A.

[0040] (1) Preparation of culture medium: take glucose, peptone 0.03g, KH 2 PO 4 0.060g, ZnSO 4 0.0001g, K 2 HPO 4 0.04g, FeSO 4 0.00005g, MnSO 4 0.005g, MgSO 4 0.050g, add 1000mL distilled water, mix well, heat to dissolve, adjust the pH to 6.0, and obtain the basic liquid medium, in which the glucose addition is 0.1g and 1.0g respectively; divide the basic liquid medium into 250mL Erlenmeyer flasks Each Erlenmeyer flask contained 30 mL of basic liquid culture medium, and 3.0 g of corn cob powder crushed to 60 mesh was added to each of them, and stirred evenly to obtain culture liquids containing different concentrations of glucose.

[0041] (2) Activation of the strains: Inoculate Pleurotus radiata (PR) and Pyriophthora filthii (DS) on potato dextrose agar plates of the same thickness that have been sterilized, respective...

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Abstract

The invention discloses a method for producing laccase by composite fermentation of white-rot fungus. The method comprises the steps of: inoculating a culture liquid with activated composite bacteria and carrying out fermentation culture at the temperature of 25-30 DEG C for 9-12 days, wherein the composite bacteria comprise at least two of phlebia radiata, dichomitus squalens and ceriporiopsis subvermispora; and the culture liquid is composed of a liquid base culture medium and crop straw powder which is at least one of corn stalk powder, corn cob powder, wheat stalk powder and rice straw powder. In the method, the corn cob powder is used as a raw material, and the combined synergic fermentation of phlebia radiate and dichomitus squalens is adopted, so that the activity of the generated laccase stably reaches 110-120 U / mL at the 12th day. The method has the characteristics that liquid fermentation is adopted, the materials are widely available, and the process is simple; aromatic compounds or heavy metal ion induction and the like are not needed and the conditions are mild; the generated laccase has high activity and good stability; and the method is a safe, environment-friendly,low-cost and efficient production method with very promising industrial prospects.

Description

technical field [0001] The invention relates to a new method for cultivating the compound fermentation of white-rot fungus with synergistic high-yield laccase, and uses different crop stalks as fermentation substrates, including corn stalks, corn cobs, wheat stalks and straw stalks, belonging to biological engineering and microbial fermentation. Background technique [0002] Laccase (Laccase, E.C.1.10.3.2) is a kind of copper-containing polyphenol oxidase, which was first discovered from the sap of sumac tree, and later found that it also widely exists in fungi, especially in white rot fungi. White rot fungi are thought to be the only organisms capable of degrading the entire woody component. Laccase uses molecular oxygen as the final electron acceptor, can widely oxidize polyphenols, methoxyphenols, diamines and other phenolic and aromatic compounds, can oxidize phenolic substances into quinones, and can catalyze the oxidation of aromatic rings. The cleavage of branched C...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12R1/645
Inventor 陈启和董亚晨何国庆傅明亮冯宇
Owner ZHEJIANG UNIV
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