Aspergillusniger strain and application thereof

A technology of Aspergillus niger strain and Aspergillus niger, which is applied in the field of enzyme engineering, can solve problems such as low fermentation enzyme activity, and achieve the effects of stable yield, easy operation and low cost

Active Publication Date: 2010-12-01
CHINA PETROLEUM & CHEM CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the above-mentioned literature has put forward some reports related to the production of β-glucosidase by fermentation, the fermentative enzyme activity is generally low

Method used

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  • Aspergillusniger strain and application thereof
  • Aspergillusniger strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0031] Slope seed medium: add 20g of glucose and 20g of agar to the infusion juice made from 200g of potatoes, add tap water to make the volume to 1000ml, and the pH is natural. This slant is used for strain activation, preservation and test tube slant seeds, etc.

[0032] Screening double layer Congo red medium:

[0033] K H 2 PO 4 1g, (NH 4 ) 2 SO 4 0.7g, MgSO 4 0.3g, CaCl 2 0.15g, 0.5ml Mandel’s Nutrient Solution——constant volume to 500ml…recorded as solution (1)

[0034] The lower layer is 200ml solution (1)+4g agar

[0035]The upper layer is 1g of CMC, 0.1g of sodium deoxycholate, and 0.01g of Congo red, which are dissolved in 100ml of solution (1), and 1g of agar is added.

[0036] The initial bacteria were derived from the activated sludge of the wastewater treatment plant. Dilute the activated sludge, suspend it in sterile water, add fermentation medium, and incubate at 30°C for 72-84h. Take an appropriate amount of culture solution, press 10 -4 , 10 -...

example 2

[0038] Nitrite mutagenesis: 0.2 mol / L acetic acid and 0.2 mol / L sodium acetate were formulated into acetic acid-sodium acetate buffer solution with pH 4.4 at 63:37 (V / V), and the slant was mixed with acetic acid-sodium acetate buffer solution Aspergillus niger T2 spores on the surface were washed to make acetic acid spore suspension. Add 0.1mol / L NaNO 2 Solution 2mL, after mutagenesis for 5min and 10min, add 2mL 0.7mol / L Na 2 HPO 4 solution as a terminator. Take 0.05mol / L NaNO with the same operation 2 Perform mutagenesis. The bacterial solution after the above treatment and the blank control were serially diluted, and diluted to the original concentration of 10 -3 、10 -4 、10 -5 0.2mL each of the solution was coated on a plate, numbered and placed in a 30°C incubator. After 72 hours of constant temperature cultivation, a single colony with a large transparent circle was picked out, and a liquid fermentation experiment was carried out in a conical flask to determine the ...

example 3

[0042] Preparation of fermentation medium: 4.95g corncobs, 3.5g yeast powder, KH in a 500ml Erlenmeyer flask 2 PO 4 0.4g, MgSO 4 0.06g, CaCl 2 0.06g, trace elements: FeSO 4 1mg, MnSO 4 0.32mg, ZnSO 4 0.2mg, add water to 200ml, pH 5.0.

[0043] Seed preparation: put the above-mentioned culture medium in the Erlenmeyer flask (500ml), sterilize at 121° C. for 30 minutes, inoculate Aspergillus niger T2 of the present invention at 10% after cooling, and cultivate at a constant temperature of 28-33° C. for 24 hours.

[0044] Liquid state fermentation: sterilize the above medium at 121°C for 30 minutes, inoculate the prepared seeds into the medium at 10% after cooling, cultivate for 72 hours at 30°C with a rotation speed of 200 rpm. The test results of the above-mentioned Erlenmeyer flasks repeated for 4 batches showed that the average enzyme activity of β-glucosidase reached 2.46U / ml.

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Abstract

The invention discloses an aspergillusniger strain for producing beta-glucuroide. Aspergillusniger T2 is preserved in the China General Microbiological Culture Collection Center (CGMCC) on 26th September, 2008 with the preservation number of CGMCC No.2715. The aspergillusniger T2 strain is the aspergillusniger strain which is obtained by performing mutagenesis with ultraviolet rays and nitrous acid and screening by using a Congo red-carboxymethylcellulose (CMC) double-layer flat plate, has high enzymatic activity and is used for producing the beta-glucuroide. The aspergillusniger T2 strain is applied to the production of the beta-glucuroide. The beta-glucuroide produced by performing liquid state fermentation on corn cob, yeast powder and the like serving as raw materials and the aspergillusniger T2 strain serving as the strain has the characteristics of enzymatic activity, simple production method and the like.

Description

technical field [0001] The invention relates to an Aspergillus niger strain and application thereof, in particular to an Aspergillus niger strain for producing β-glucosidase, a production method and application thereof, and belongs to the field of enzyme engineering. technical background [0002] In the process of hydrolyzing natural cellulose into glucose, it must rely on the synergistic action of three enzymes: endoglucanase, exoglucanase and β-glucosidase. Among them, β-glucosidase hydrolyzes cellobiose or cellooligosaccharides into glucose molecules, which is the rate-limiting factor in this process, and its molecular weight is about 76ku. In the process of using cellulase to hydrolyze cellulose, the deficiency of β-glucosidase leads to the accumulation of cellobiose, and cellobiose will form a strong feedback inhibition on the catalysis of cellulase. Therefore, enhancing the activity of β-glucosidase is one of the key measures to improve the hydrolysis rate of cellulos...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C12N9/42C12R1/685
Inventor 唐开宇张全佟明友尹佩林
Owner CHINA PETROLEUM & CHEM CORP
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