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Method for Aspergillus oryzae to secrete lignin to prepare peroxidase

A technology of peroxidase and lignin, which is applied in the crude product field of lignin peroxidase, and can solve problems such as unseen

Inactive Publication Date: 2013-12-18
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Aspergillus oryzae is a feed-grade microbial additive strain that can be directly fed to animals recognized by the U.S. Food and Drug Administration (FDA), the American Feed Association (AAFCO), and the Ministry of Agriculture of my country, but no lignin peroxide secretion by Aspergillus oryzae has been seen Enzyme patents and related literature reports

Method used

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  • Method for Aspergillus oryzae to secrete lignin to prepare peroxidase

Examples

Experimental program
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Effect test

Embodiment 1

[0029] 1. Production of bacteria on test tube slant

[0030] Insert a ring of Aspergillus oryzae (Aspergillus oryzae) into the newly configured potato sucrose medium ("Industrial Microbiology Experimental Technology Manual" edited by Zhuge Jian and Wang Zhengxiang, 1994, page 367) (China Common Microbial Cultures Preservation Management Center, CGMCC5992), 26°C, incubation time 50 hours; store at 4°C for later use.

[0031] 2. Production of cultured strains in liquid shake flasks

[0032]Accurately weigh 5 grams of glucose, 0.1 grams of ammonium tartrate, 0.2 grams of benzyl alcohol, 0.2 grams of magnesium sulfate, 0.5 grams of Tween 80, 2 grams of potassium dihydrogen phosphate, 3 grams of phthalic acid buffer, 1000 mL of water, pH 5 , divided into 250mL triangular flasks, 50 grams per bottle, a total of 20 bottles, sterilized at 120 °C for 40 minutes; after cooling, each bottle was inserted into a ring of Aspergillus oryzae strains preserved at 4 °C, at 25 °C, 150 rpm, Cul...

Embodiment 2

[0042] 1. Production of bacteria on test tube slant

[0043] Insert a ring of Aspergillus oryzae into the newly configured potato sucrose medium ("Industrial Microbiology Experimental Technology Manual" edited by Zhuge Jian and Wang Zhengxiang, 1994, page 367), at 30°C, and cultivate for 100 hours ; Store at 7°C for later use.

[0044] 2. Production of cultured strains in liquid shake flasks

[0045] Accurately weigh 150 grams of glucose, 250 grams of ammonium tartrate, 15 grams of benzyl alcohol, 0.4 grams of magnesium sulfate, 4 grams of Tween 80, 5 grams of potassium dihydrogen phosphate, 20 grams of phthalic acid buffer, 10 L of water, pH 6, Divide into 250mL triangular flasks, 100 grams per bottle, 100 bottles in total, sterilize at 130°C for 30 minutes; after cooling, insert a ring of Aspergillus oryzae strains preserved at 7°C, and cultivate 50 hours;

[0046] 3. Preparation of first-level liquid strains

[0047] Accurately weigh 2000 grams of glucose, 200 grams of ...

Embodiment 3

[0055] 1. Production of bacteria on test tube slant

[0056] Insert a ring of Aspergillus oryzae into the newly configured potato sucrose medium ("Industrial Microbiology Experimental Technology Manual" edited by Zhuge Jian and Wang Zhengxiang, 1994, page 367), at 35°C, and cultivate for 144 hours ; Store at 10°C for later use.

[0057] 2. Production of strains cultured in liquid shake flasks

[0058] Accurately weigh 300 grams of glucose, 50 grams of ammonium tartrate, 30 grams of benzyl alcohol, 10 grams of magnesium sulfate, 100 grams of Tween 80, 90 grams of potassium dihydrogen phosphate, 180 grams of phthalic acid buffer, 10 L of water, pH7.5 , divided into 250mL triangular flasks, each bottle 120 mL, a total of 80 bottles, sterilized at 140 ° C for 20 minutes; after cooling, each bottle was inserted into a ring of Aspergillus oryzae strains stored at 10 ° C, at 35 ° C, 150 rpm, Cultivate for 72 hours;

[0059] 3. Preparation of first-level liquid strains

[0060] Ac...

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Abstract

Relating to the field of biological engineering, the invention provides to a method for Aspergillus oryzae to secrete lignin to prepare peroxidase. The production process includes: subjecting straw to crushing, pre-wetting and other pre-treatment technologies, and subjecting Aspergillus oryzae to shaking culture, first class seed culture, and mixing with straw to undergo solid state fermentation so as to obtain a lignin peroxidase-containing straw fermentation product with enzyme activity of 3-800U / g dry fermentation product, and carrying out water extraction, molecular interception and freeze drying on the fermentation product so as to obtain a crude enzyme preparation with enzyme activity of 900-8000U / g dry crude product. The fermentation process is simple, has no pollution to the environment, and causes no resource waste. The lignin peroxidase prepared by the method can be used to degrade lignin macromolecular structures, release small molecule substances, and improve the digestion utilization rate of lignin-containing straw, thus being helpful to improve the feeding value of straw.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to fermenting straw culture medium without high temperature and high pressure treatment to prepare lignin peroxidase product, extracting the product with water, and lyophilizing the extract to obtain lignin peroxidase crude products. Background technique [0002] my country is a large agricultural country, with an annual output of more than 600 million tons of crop straw, except for a small amount that is used in the textile and papermaking industries, but most of it is dumped into the environment in the form of accumulation and incineration, which not only wastes resources but also causes great harm to the environment . [0003] Straw contains more than half of the energy accumulated by plant photosynthesis. It is rich in raw sugar, crude protein and other ingredients that are beneficial to livestock and poultry. As a non-competitive feed resource, as long as it can be processed a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/08C12R1/69
Inventor 张志才陈钧任晓锋李佳少黄达明
Owner JIANGSU UNIV
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