Aspergillus awamori and application thereof in preparation of tannase and degradation of tannin

A kind of Aspergillus awamori and tannin technology, applied in the field of tannin degradation, can solve the problems of high corrosion resistance of equipment, difficulty in industrialization, limited application, etc., achieve wide pH adaptability, realize industrial production, and not easy to material denaturation Effect

Active Publication Date: 2021-04-13
NANJING NORMAL UNIVERSITY
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  • Claims
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Problems solved by technology

Chemical degradation is oxidative degradation or high sulfitation by using oxidants such as strong acid or strong alkali in high temperature environment. Its degradation efficiency is high, but it is easy to cause environmental pollution and has high requirements for corrosion resistance of equipment, so it is not easy to apply to industrialization
The biodegradation method mainly uses the key restriction enzyme tannase to degrade tannins, which can act on hydrolyzed tannins, complex tannins, and esters formed by gallic acid and alcohol. The degradation is specific, and the production process is green and pollution-free. However, the enzyme activity of commercial enzymes currently used for biodegradation is about 200U / g, and the price is relatively high, which limits its application in industry

Method used

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  • Aspergillus awamori and application thereof in preparation of tannase and degradation of tannin

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preparation example Construction

[0046] According to the present invention, the method for degrading tannin preferably includes: contacting the bacterium agent with the sample containing tannin, the bacterium agent being the fermentation product of the Aspergillus awamori; wherein, the fermentation product of the Aspergillus awamori The preparation method comprises the following steps: performing seed culture and fermentation culture of the Aspergillus awamori, obtaining fermentation cells through solid-liquid separation II, and performing solid-liquid separation III on the fermentation cells after cell crushing to obtain clear liquid.

[0047] Preferably, the cell breaking process includes: washing the fermented cells with a buffer solution, then mixing with the buffer solution and performing ultrasonic disruption. Exemplarily, the solid-liquid separation II is separated by filtration. Exemplarily, it can be filtered by sterilized gauze; the buffer is citric acid-sodium citrate buffer, and the ultrasonic crus...

Embodiment 1

[0053] S1, sampling: the sample is collected from the soil sample under the tea tree in Liuzhou, Guangxi (25°13' north latitude, 108°54' east longitude);

[0054] S2. Enrichment: Take 1g of soil sample in 9mL of sterile water, add glass beads to disperse the soil sample and shake it evenly, and dilute the soil bacteria suspension to 10% by gradient dilution method. -1 -10 -5 The enrichment medium is Bengal red culture medium (the main components are Bengal red, chloramphenicol, magnesium sulfate, potassium dihydrogen phosphate, agar, glucose, peptone), absorb 200 μL of the bacterial solution after gradient dilution, and use a coating stick to Spread evenly in Bengal red medium, and carry out enrichment culture to obtain enriched strains;

[0055] S3, initial screening: select Cha's solid medium plus tannic acid (tannic acid content is 2% by weight) as the initial screening medium, the enriched strains obtained in S2 are picked and inoculated on the initial screening medium wi...

Embodiment 2

[0059] (1) Grind the dried pomegranate peel into powder, add 30 mL of ethanol-water solution with an ethanol concentration of 70% by volume to 1 g of pomegranate peel powder, vibrate for 1 hour at a temperature of 60°C and a rotational speed of 180rpm, and then shake it at a temperature of 60°C 1. After ultrasonic treatment for 1 h under the condition of ultrasonic power of 550 W, centrifuge at 8000 rpm for 5 min at room temperature to obtain the extract;

[0060] (2) The extract obtained in step (1) is removed by rotary steaming to remove ethanol until it is colloidal to obtain a rotary steamed product, and the rotary steamed product is mixed with 30 mL of pure water to obtain a sample containing tannin, and the ellagitannin in the sample is determined The content is about 7.90g / L;

[0061] (3) After activating the Aspergillus awamori obtained in Example 1, inoculate it in the seed medium, cultivate it for 48 hours at a temperature of 30° C. and a rotation speed of 220 rpm to...

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Abstract

The invention relates to a tannin degradation technology, and discloses aspergillus awamori and application thereof in preparation of tannase and degradation of tannin. Specifically, the aspergillus awamori is provided, and the preservation number of the aspergillus awamori is GDMCC No.61266. The invention also provides a microbial inoculum. The microbial inoculum contains the aspergillus awamori. The invention also provides a method for degrading tannin. The method comprises the step of contacting the aspergillus awamori and / or the microbial inoculum with a tannin-containing sample. The aspergillus awamori provided by the invention can be used for efficiently producing tannase, and the conditions of an enzymolysis reaction have wide pH adaptability, so that degradation of tannin can be efficiently catalyzed in a short time, and substance denaturation is not easily caused.

Description

technical field [0001] The invention relates to tannin degradation technology, in particular to Aspergillus awamori and its application in preparing tannase and degrading tannin. Background technique [0002] Tannin is a polyphenolic compound that widely exists in plants. Among plant phenolic substances, its content is second only to lignin, and it mostly occurs in forage trees, shrubs, legumes and grains with high nutritional value middle. Tannin can destroy and hinder the digestion and utilization of nutrients. It is a typical anti-nutritional factor. At present, its utilization is mainly in the form of small molecules such as ellagic acid, catechin, anthocyanin and its derivatives. Therefore, by degrading tannin , Obtaining low molecular weight tannins is an important way to improve the utilization rate of tannins and the fine utilization of tannins. [0003] The degradation of macromolecular tannins mainly includes chemical degradation and biological degradation. Chem...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N9/18C12R1/66
CPCC12N1/14C12N9/18C12Y301/0102
Inventor 黄和王文慧屈艳君曹家南齐雅雯韦智匀宋萍徐晴
Owner NANJING NORMAL UNIVERSITY
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