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Aspergillus fumigatus strain Bfum-5 and application thereof

A bfum-5, Aspergillus fumigatus technology, applied in fungi, microorganisms, biochemical equipment and methods, etc., can solve the problems of high cost, environmental pollution, complicated degradation conditions, etc., and achieve the effect of strong cellulose degradation

Active Publication Date: 2015-10-28
北京大地聚龙生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the previous methods have the disadvantages of high cost, cumbersome degradation conditions and easy to cause environmental pollution, while the biodegradation method mainly uses cellulase to catalyze the degradation of cellulose, which has become the main method due to its high specificity and low cost. a means of attention

Method used

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  • Aspergillus fumigatus strain Bfum-5 and application thereof
  • Aspergillus fumigatus strain Bfum-5 and application thereof
  • Aspergillus fumigatus strain Bfum-5 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1, the screening process of Aspergillus fumigatus strain Bfum-5 of the present invention

[0057] Materials and Methods

[0058] 1 Extraction method of cellulolytic bacteria

[0059] 1.1 Materials

[0060] The high-temperature compost mixture of pocket mushroom residue and duck manure (denoted by U), the high-temperature compost mixture of Pleurotus eryngii residue and duck manure (denoted by N).

[0061] 1.2 Method

[0062] dilution plate method

[0063] 1.2.1 Medium

[0064] Hutchison Filter Medium: KH 2 PO 4 1g / L, NaCl 0.1g / L, MgSO 4 0.3g / L, NaNO 3 2.5g / L, FeCl 3 0.01g / L, CaCl 2 ·6H 2 O 0.1g / L, 20g agar, several sheets of starch-removed filter paper (one for each plate).

[0065] 1.2.2 Main steps

[0066] a. Preparation of starch-removing filter paper: soak the filter paper with 1% acetic acid for a day and night, check with iodine solution to confirm that there is no starch, then rinse it with 2% sodium bicarbonate solution until neutral, a...

Embodiment 2

[0095] Example 2 Molecular Identification of Cellulolytic Bacteria CZ5-5

[0096] Materials and Methods

[0097] 1. Strain DNA extraction

[0098](1) Under sterilized conditions, pick an appropriate amount of spores with a sterilized pipette tip in the ultra-clean workbench and place them in a 2mL sterile centrifuge tube, add 0.8mL extraction buffer (1% CTAB; 100mM Tris-HCl pH 8.0; 20mM EDTA pH 8.0; 700mMNaCl8.0) and 20μL proteinase K, mix thoroughly. 37°C, 30min, mix every 10min.

[0099] (2) Add 160 μL of 10% SDS and mix thoroughly. Water bath at 65°C for 1 hour, and mix well every 15 minutes.

[0100] (3) Add an equal volume of chloroform, mix thoroughly, and centrifuge at 11,000 rpm at 4°C for 10 min.

[0101] (4) Take 700 microliters of supernatant to a 2mL centrifuge tube;

[0102] (5) Repeat steps 3 and 4;

[0103] (6) Add 0.6 times the volume of isopropanol to mix evenly to precipitate, and centrifuge.

[0104] (7) Add 700ul of 75% ethanol, mix well and centr...

Embodiment 3

[0123] Embodiment 3, verify the cellulose degrading effect of selected single strain

[0124] 1. Filter paper degradation test method

[0125] All the single strains obtained after screening, and some strains with strong ability to decompose cellulose obtained from the CMC-Na hydrolysis circle test were further screened by the filter paper degradation test.

[0126] 1 medium

[0127] Filter paper medium: starch-removed filter paper strips 1*3cm several sheets (one per bottle), (NH4)2SO42g / L, KH2PO41g / L, MgSO4·7H2O 0.5g / L, NaCl 0.5g / L, NaNO31g / L, FeCl30.01g / L, CaCl2·2H2O 0.2g / L, peptone 0.5g / L, yeast extract 0.5g / L, pH7.2 or so.

[0128] 2 main steps

[0129] a. Select the bacterial species, and do 3 repetitions for each bacterial species, including 1 sterile water control group and 2 filter paper medium experimental groups. Cultivate for 10 days in a shaking box at 35°C and 150r / min.

[0130] b. From the fourth day, observe once every other day to see if the filter paper ...

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Abstract

The invention relates to an aspergillus fumigatus strain Bfum-5 and its application and belongs to the field of microbial ecology. Preservation number of the aspergillus fumigatus strain Bfum-5 is CGMCC No.10929. The invention also provides a product and microbial inoculums used for degrading cellulose with the aspergillus fumigatus strain Bfum-5 as an active ingredient and a method for degrading cellulose based on the strain Bfum-5. Meanwhile, the invention provides an application of the strain Bfum-5 in the preparation of a product for degrading cellulose. Through CMC-Na hydrolyzation circle determination, filter paper degradation experiments and cellulase enzyme activity determination, it has been confirmed that the aspergillus fumigatus strain Bfum-5 has very strong capability of degrading cellulose.

Description

technical field [0001] The invention belongs to the field of microbial ecology, and in particular relates to an Aspergillus fumigatus strain Bfum-5 and its application. Background technique [0002] Cellulose is the most abundant and widely distributed natural organic matter in nature. It is the main component of the mature cell wall of cotton, wood and other higher plants. It is considered as the "basic substance of the cell", and billions of tons are produced by the action of light every year. Cellulose accounts for about 40-45% in dry stems of gramineous plants such as reeds, bamboos, and straws, about 40-50% in wood, and the highest content in cotton, reaching 95-99%. The use of cellulose can be traced back to the invention of papermaking two thousand years ago. With the intensification of the energy crisis, the development and utilization of bio-energy has gradually been paid attention to, and cellulose, as the cheapest and most abundant renewable energy on the earth,...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P1/02C12R1/68
Inventor 刘悦秋刘克峰王树涛高程达于峰张泽
Owner 北京大地聚龙生物科技有限公司
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