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Myceliophthora thermophila ferment as well as preparation method and application thereof

A technology of Myceliophthora thermophila and fermented products, which is applied in the field of fermented Myceliophthora thermophila and its preparation and application, and can solve the problems of easy inactivation and other problems

Active Publication Date: 2014-01-08
ANHUI NEW SIMON BIOTECH CO LTD
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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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  • Myceliophthora thermophila ferment as well as preparation method and application thereof
  • Myceliophthora thermophila ferment as well as preparation method and application thereof
  • Myceliophthora thermophila ferment as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1, the application of Myceliophthora thermophila in the production of cellulase

[0033] 1. Seed cultivation

[0034] Inoculate Myceliophthora thermophila on the PDA slant and culture at 45°C for 5 days.

[0035] 2. Fermentation

[0036] Adopt 250ml Erlenmeyer flask, and 50ml fermentation medium is housed in each Erlenmeyer flask.

[0037] Dig the seed slant (approximately 0.5×1 cm) into the fermentation medium, culture at 45°C, 180r / min shaking (radius 13mm) for 5 days, then centrifuge at 5000g for 20min, and collect the supernatant.

[0038] Fermentation medium (pH=5): take 50g of microcrystalline cellulose, 27g of corn steep liquor, 5g of ammonium sulfate, 6g of potassium dihydrogen phosphate, 2.5g of calcium carbonate, 1g of magnesium sulfate, 2.5g of glycerol and tween-802g, dissolve in water and make up to 1L with water.

[0039] 3. Determination of Enzyme Activity

[0040] Determination of enzyme activity (reaction temperature is 50°C, reaction pH...

Embodiment 2

[0042] Embodiment 2, optimization of the fermentation conditions of Myceliophthora thermophila

[0043] 1. The effect of different carbon sources on the production of cellulase

[0044] 1. Seed cultivation

[0045] Inoculate Myceliophthora thermophila on the PDA slant and culture at 45°C for 5 days.

[0046] 2. Fermentation

[0047] Adopt 250ml Erlenmeyer flask, and 50ml fermentation medium is housed in each Erlenmeyer flask.

[0048] Dig the seed slant (approximately 0.5×1 cm) into the fermentation medium, culture at 45°C, 180r / min shaking (radius 13mm) for 5 days, then centrifuge at 5000g for 20min, and collect the supernatant.

[0049] Fermentation medium (pH=5): 50g of carbon source (microcrystalline cellulose, corncob or rice straw powder), 27g of corn steep liquor, 5g of ammonium sulfate, 6g of potassium dihydrogen phosphate, 2.5g of calcium carbonate, 1g of magnesium sulfate, glycerin 2.5g and Tween-802g, dissolve in water and dilute to 1L with water.

[0050] 3. Det...

Embodiment 3

[0082] Embodiment 3, the enzymatic property of cellulase

[0083] 1. Optimum reaction temperature

[0084] Take step 2 of Example 1 to obtain the supernatant, and detect the enzyme activity with reference to the enzyme activity assay method of step 3 of Example 1. The only difference is that the following reaction temperatures are used respectively: 30°C, 40°C, 50°C, 60°C and 70°C ℃.

[0085] For supernatant results, see Figure 5 (The unit of the ordinate is U / mL). It has the highest enzyme activity at 60°C.

[0086] 2. Optimum reaction pH

[0087] Get the supernatant obtained in Step 2 of Example 1, and detect the enzyme activity with reference to the enzyme activity assay method in Step 3 of Example 1. The only difference is that the following buffer solution is used to replace "pH4.8, 0.05M disodium hydrogen phosphate-lemon Acid Buffer Buffer": 0.05M disodium phosphate-citrate buffer at pH 3.0, 4.0, 5.0 or 6.0.

[0088] For supernatant results, see Image 6 (The uni...

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Abstract

The invention discloses a myceliophthora thermophila ferment as well as a preparation method and an application thereof. The invention provides a method for preparing the ferment, wherein the method comprises the following steps: inoculating the myceliophthora thermophila ACCC No. 30572 into a fermenting culture medium, vibrating and culturing for 5-9 days under the condition of 40-55 DEC and 100-300rpm; the fermenting culture medium is prepared by the method comprising the following steps: mixing a carbon source, a nitrogen source, ammonium sulfate, monopotassium phosphate, calcium carbonate, magnesium sulfate, glycerol and tween-80 with water, thus obtaining the fermenting culture medium, wherein the concentration of the carbon source is 25-60g / L, the concentration of the nitrogen source is 10-40g / L, the concentration of the ammonium sulfate is 1-10g / l, the concentration of the monopotassium phosphate is 1-10g / L, the concentration of the calcium carbonate is 1-4g / L, the concentration of the magnesium sulfate is 0.5-4g / L, the concentration of the glycerol is 1-4g / L, and the concentration of the tween-80 is 0.5-3g / L. The ferment provided by the invention has the activity of cellulose, has excellent temperature stability and pH (potential of hydrogen) stability, namely, has higher enzyme activity under a high-temperature condition, and has higher enzyme activity within a wider pH scope.

Description

technical field [0001] The invention relates to a fermented product of Myceliophthora thermophila and its preparation method and application. Background technique [0002] Cellulase is a complex enzyme system that can degrade cellulose into glucose, including: (1) 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 cellobiohydrolases (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 textile industry and bioenergy), cellulase has become a hot spot in enzyme engi...

Claims

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Application Information

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IPC IPC(8): C12N9/42C12R1/645
CPCC12N9/2437C12N9/2445C12Y302/01004C12Y302/01021C12Y302/01091
Inventor 杨敬钞亚鹏钱世钧陈树林马延和张国青石家骥
Owner ANHUI NEW SIMON BIOTECH CO LTD
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