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Thermophilic Aspergillus funigatus strain and application thereof in production of cellulase

A technology of cellulase and Aspergillus fumigatus, applied to a strain of Aspergillus fumigatus and its application in the production of cellulase, which can solve the problems of easy inactivation

Active Publication Date: 2015-06-10
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the cellulases produced by the above strains need to perform better at temperatures below 50°C, but are easily inactivated at higher temperatures

Method used

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  • Thermophilic Aspergillus funigatus strain and application thereof in production of cellulase
  • Thermophilic Aspergillus funigatus strain and application thereof in production of cellulase
  • Thermophilic Aspergillus funigatus strain and application thereof in production of cellulase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1, the acquisition and identification of Aspergillus fumigatus HS-1

[0020] 1. Acquisition of strain HS-1

[0021] Soil samples: collected from humus soil in Tengchong, Yunnan.

[0022] Plate screening medium: cellulose powder 10g / L, glucose 20g / L, potato juice 1L, agar 20g / L; pH natural; sterilized at 115°C for 15 minutes; add 15ml of the medium to a plate with a diameter of 9cm.

[0023] Prepare the soil sample into a suspension with a concentration of 10-2-10-5, take 0.1ml to smear the dish, and put it in a 45°C incubator for 2-3 days (see photos Figure 4 , the right picture is the control, and the left picture can see a very clear transparent circle) After that, a strain producing acid high-temperature cellulase was obtained, and it was named strain HS-1.

[0024] 2. Identification of strain HS-1

[0025] 1. Identification of morphological characteristics

[0026] The strain grows rapidly on the PDA medium, and it is white fluff at the beginning, and t...

Embodiment 2

[0031] Embodiment 2, the application of Aspergillus fumigatus HS-1 in the production of cellulase

[0032] 1. Seed cultivation

[0033] Inoculate Aspergillus fumigatus HS-1 on PDA slant and culture at 45°C for 5 days.

[0034] 2. Fermentation

[0035] Digging pieces from the inclined surface of the seeds were inserted into the fermentation medium, cultured on a rotary shaker at 45°C and 180r / min for 5 days, and then centrifuged at 5000g for 20min to collect the supernatant.

[0036] Fermentation medium: Take 50g of corncobs, 5g of yeast powder, 2.8g of ammonium sulfate, 4g of potassium dihydrogen phosphate, 0.9g of calcium chloride, and 0.9g of magnesium sulfate, dissolve them in water and dilute to 1L with water; pH4.5.

[0037] 3. Determination of Enzyme Activity

[0038] Determination of enzyme activity (reaction temperature is 50°C, reaction pH is 4.8): take 50mg filter paper (Xinhua No. 1) as substrate, add 1ml pH4.8, 0.05mM citric acid-sodium citrate buffer, bathe in wa...

Embodiment 3

[0041] Embodiment 3, the enzymatic property of cellulase

[0042] 1. Optimum reaction temperature

[0043] Concentrate the supernatant obtained in Step 2 of Example 2 to obtain a concentrated solution, and detect the enzyme activity of the concentrated solution with reference to the enzyme activity assay method in Step 3 of Example 2. The only difference is that the following reaction temperatures are used respectively: 30°C, 40°C , 50°C, 60°C and 70°C.

[0044] The result is as figure 1 shown. The cellulase produced by Aspergillus fumigatus HS-1 had the highest enzyme activity at 60°C.

[0045] 2. Optimum reaction pH

[0046] Concentrate the supernatant obtained in Step 2 of Example 2 to obtain a concentrated solution, and detect the enzyme activity of the concentrated solution with reference to the enzyme activity assay method in Step 3 of Example 2. The only difference is that the following buffer solutions are used to replace "pH4.8, 0.05mM citric acid-sodium citrate ...

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Abstract

The invention discloses a thermophilic Aspergillus funigatus strain and application thereof in production of cellulase. The collection number of the Aspergillus funigatus HS-1 provided by the invention is CGMCC No.7563. The invention also protects application of the Aspergillus funigatus HS-1 in preparation of cellulase. The invention also protects a cellulase preparation method which comprises the following steps: fermenting the Aspergillus funigatus HS-1, and collecting the fermentation product to obtain the cellulase. The strain provided by the invention can be induced to produce the cellulase in a culture medium taking a corn cob as the unique carbon source, and the maximum enzyme-producing activity is 1.004U / mL. The optimum pH value of the enzyme is 5.0, and the enzyme is active when the pH value is 3-8; the optimum temperature is 60 DEG C; and the enzyme has the characteristics of high-temperature cellulase.

Description

technical field [0001] The invention relates to a strain of Aspergillus fumigatus and its application in producing cellulase. Background technique [0002] Cellulase is a compound enzyme system that can degrade cellulose into glucose. It mainly consists of three components, namely: (1) endoglucanase (endo-β-1,4-D- glucanase, EG, EC3.2.1.4), also known as CMC enzyme; (2) exo-β-1,4-glucanase (EC3.2.1.91) or cellobiohydrolase (cello -biohydrolases, CBH); (3) Glucosidase (β-1,4-D-glucosidase, BG, EC3.2.1.21), also known as salicinase. Each of the above three enzymes has multiple isozymes. Endonuclease cuts long-chain cellulose from the inside, exonuclease cuts off disaccharides or oligosaccharides from the reducing or non-reducing ends of cellulose chains, and glucosidase hydrolyzes cellobiose or oligosaccharides into glucose. With the wide application of cellulase in industry (such as: food, feed, brewing, papermaking, especially in the textile industry and bioenergy), cellu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/42C12R1/68
Inventor 钞亚鹏钱世钧杨敬陈树林马延和石家骥张国青
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI