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Method for producing ligninase by using edible mushroom stick

A technology of ligninase and edible fungus, applied in the direction of biochemical equipment and methods, enzymes, enzymes, etc., can solve the problems of unfavorable large-scale application, low ligninase activity, high separation and purification costs, etc., to shorten the production of enzymes time, improve the environment, and reduce production costs

Inactive Publication Date: 2015-06-10
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has also been reported that ligninase is directly extracted from edible fungus residues. However, due to the low activity of ligninase in the residues cultured without enzyme-inducing solution, high separation and purification costs are required, and the economic benefits are low. Facilitate large-scale application
[0004] So far, there has not been any technology that uses edible fungus sticks as raw materials to add enzyme-producing inducing solution for secondary culture to produce ligninase

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 ligninase activity assay method

[0025] Take 10 μl of the crude enzyme solution obtained in step 4, divide it into 10 centrifuge tubes of 1 ml, centrifuge at 12000 r / min for 10 minutes, and take the supernatant to measure ligninase activity.

[0026] Manganese peroxidase (Mnp) enzyme activity assay method: at 30°C, the reaction system contains 50mmol, 2.0mL of tartrate-sodium tartrate buffer solution with pH 5.0, 0.8mL of 1.6mmol / L manganese sulfate solution, and 0.1 mL, 20mmol / L DMP 25μL, add 1.6mmol / L hydrogen peroxide 75μL to start the reaction. The change in absorbance at 470 nm was measured during the first 1 minute of the reaction. One unit of enzyme activity is defined as the amount of enzyme required to produce 1 μmol of product per minute.

[0027] Lignin peroxidase (Lip) enzyme activity assay method: with reveratrol as substrate, 2.3mL of tartrate-sodium tartrate buffer solution with pH 3.5, 100μL of 10mmol / L reveratrol, 0.4mmol / L peroxide Hyd...

Embodiment 2

[0030] Take 10kg of Pleurotus ostreatus mushroom sticks, crush them to obtain the chaff, add 15L of enzyme-producing inducing solution, and culture at 28°C for 3 days, soak the obtained system in 150L of distilled water, oscillate, and filter to obtain the crude enzyme solution and fungus chaff. Continue to add 15L of enzyme-producing induction solution to the remaining fungus chaff, incubate at 28°C for 3 days, soak the obtained system in 150L of distilled water, oscillate, filter to obtain crude enzyme solution and fungus chaff, and cycle like this for 3 times to obtain 450L of For the crude enzyme solution, the ligninase activity measured according to Example 1 is as follows: 4 U of manganese peroxidase activity, 3 U of lignin peroxidase activity, and 22 U of laccase activity.

Embodiment 3

[0032]Take 10kg of Agaricus bisporus sticks, crush them to obtain chaff, add 25L of enzyme-producing inducing solution, and incubate at 22°C for 4 days, soak the obtained system in 150L of distilled water, oscillate, and filter to obtain crude enzyme solution and chaff. Continue to add 25L of enzyme-producing induction solution to the remaining fungus chaff, incubate at 22°C for 4 days, soak the obtained system in 150L of distilled water, oscillate, filter to obtain crude enzyme solution and fungus chaff, and cycle like this for 3 times to obtain 450L of The ligninase activity of the crude enzyme solution measured according to Example 1 is as follows: 2 U of manganese peroxidase activity, 1 U of lignin peroxidase activity, and 17 U of laccase activity.

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Abstract

The invention discloses a method for preparing ligninase from mushroom residue. The method comprises the specific steps: crushing the mushroom stick to obtain mushroom residue, adding enzyme-production induced liquid, cultivating under proper condition, recycling crude enzyme and repeatedly adding the enzyme-production induced liquid; the enzyme activity of obtained paint enzyme is 1-10U, the manganese peroxidase is 1-10U, and the lignin peroxidase is 10-30U. compared with other prior art related to the ligninase production method, the method saves the mushroom growth process, saves the energy source and the material, the waste is utilized, the environment is improved, and the cost for preparing the ligninase is effectively reduced.

Description

technical field [0001] The invention relates to a method for producing ligninase, in particular to a method for producing ligninase by using edible mushroom sticks as raw materials. technical background [0002] Lignin-degrading enzymes is a general term for a group of enzymes that can catalyze the oxidative degradation of lignin, including laccase (abbreviated as Lac), manganese peroxidase (abbreviated as Mnp) and lignin peroxidase (abbreviated as Lip). Laccase is the system name of diphenol oxidoreductase, which acts on the oxidation reaction of o-ketol and p-ketol, aminophenol and diamine; manganese peroxidase is the system name of divalent manganese hydrogen peroxide oxidoreductase , to catalyze the oxidation reaction of divalent manganese to trivalent manganese; lignin peroxidase, the system name is lignin hydrogen peroxide oxidoreductase, which catalyzes the oxidation reaction of lignin. Studies have found that they have a strong ability to degrade many compounds con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/08C12N9/02
CPCC12N9/0061C12N9/0065C12Y110/03002C12Y111/01C12Y111/01013C12Y111/01014
Inventor 廖祥儒张永田乔鹏蔡宇杰管政兵
Owner JIANGNAN UNIV
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