Aspergillus niger HC306 and application of aspergillus niger HC306 to prepare naringenin through naringin conversion

A technology of Aspergillus niger and catalyst, which is applied in the field of preparation of naringenin, can solve the problems of few by-products, low conversion yield, poor specificity, etc., and achieves low by-products, high conversion yield and stable batch Effect

Active Publication Date: 2016-08-10
ZHEJIANG SHUREN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a microbial strain producing naringinase—Aspergillus niger (Aspergillus niger) HC306, and its application in transforming naringin to prepare naringenin, which can overcome the exis

Method used

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  • Aspergillus niger HC306 and application of aspergillus niger HC306 to prepare naringenin through naringin conversion
  • Aspergillus niger HC306 and application of aspergillus niger HC306 to prepare naringenin through naringin conversion
  • Aspergillus niger HC306 and application of aspergillus niger HC306 to prepare naringenin through naringin conversion

Examples

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Example Embodiment

[0023] Example 1: Enrichment and isolation of transformed strains

[0024] Add about 20g of crushed fresh grapefruit peel to a 250mL Erlenmeyer flask and incubate at 28°C for 5 days. Dilute the enriched material covered with mold with sterile water 1×10 6 After doubling, spread it on a PDA plate medium, culture at 28°C for 4 days, pick out mold colonies with different colors and shapes, transfer to PDA slant medium, and place it at 28°C for 3 days to obtain a spore-rich slant strain Nine strains were numbered (HC301~HC309) and stored in a refrigerator at 4°C for later use.

[0025] Both the plate medium and the slope medium are potato dextrose agar medium (PDA), which are prepared according to the following composition and method: wash the potatoes, peel them and cut into small pieces, weigh 200g, add 1000mL tap water, boil for 30min, 4 Filter with a layer of gauze to remove the residue, make up the filtrate to 1000mL, add 20g of sucrose, 18g of agar, natural pH (measured 6.5), he...

Example Embodiment

[0026] Example 2: Screening and classification of transformed strains

[0027] Dip the slant spores of each strain obtained in Example 1 with cotton swabs 3 times, insert them into 50mL fermentation medium, culture for 4 days at 28°C and 200r / min constant temperature shaking for enzyme production, 50mL fermentation broth is pumped with Buchner funnel Filter, the collected filtrate is the crude enzyme solution, about 40mL. Take 25mL of crude enzyme solution and place it in a 150mL Erlenmeyer flask. Dissolve 0.25g of naringin in 25mL of pH 4.0 phosphate buffer. After mixing the two to form a transformation system (total volume: 50mL), shake at 40℃ and 200r / min. Transform under the conditions for 2h; after the transformation reaction, take 5mL of the transformation solution, centrifuge at 8000g for 10min, filter through a 0.45μm microporous membrane, and analyze the concentration of naringenin in the sample by high performance liquid chromatography (HPLC).

[0028] HPLC method was us...

Example Embodiment

[0038] Example 3: Application of Aspergillus niger HC306 strain transforming naringin to naringenin 1

[0039] Taking the Aspergillus niger HC306 screened in Example 2 as the enzyme-producing strain, the crude enzyme solution prepared by fermentation was expanded and cultured by fermentation to treat naringin. The molar conversion yield of naringenin was slightly higher than that in Example 2. Repeat There is no significant difference in the results of experiment 3, which indicates that the enzyme-producing naringin of this strain is stable to produce naringenin. The specific process steps are as follows:

[0040] (1) The Aspergillus niger HC306 slant strain stored in the refrigerator at 4°C was inoculated into fresh PDA slant medium, and the slant was cultured at 28°C for 3 days. The composition and preparation method of the PDA inclined plane culture medium are the same as in Example 1;

[0041] (2) Dip a cotton swab into the step (1) After activation and culture, the Aspergillus ...

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Abstract

The invention discloses aspergillus niger HC306 and an application of aspergillus niger HC306 to prepare naringenin through naringin conversion. The aspergillus niger HC306 is preserved at Guangdong Microbiology Culture Center and is provided with the accession number of GDMCC No: 60026. The preservation date is Mar. 21<st>, 2016, and the preservation address is the fifth floor, No. 59 building, No. 100 courtyard, Xianliezhonglu, Guangzhou, with the zip code being 510075. The aspergillus niger HC306 is free of toxicity or harmlessness, is safe to use, fast in growth, and good in microbial resistance, and is stable in batches. A fermentation matrix is simple in ingredient and low in market price, and therefore the production cost of naringinase is relatively low. A crude enzyme liquid containing naringinase can be straight applied to the preparation of phloretin, thereby avoiding the separation and purification step of an enzyme and assisting in the reduction of the production cost. The conversion yield of naringenin is high and can reach as high as 93.4%. The preparation method of aspergillus niger HC306 has advantages of fewer by-products and easy extraction of products.

Description

(1) Technical field [0001] The invention relates to a method for preparing naringenin, in particular to a strain of Aspergillus niger HC306 and the application of the strain in transforming naringenin to prepare naringenin. (2) Background technology [0002] Naringenin (naringenin), also known as citrus, naringenin and naringenin, chemical name 4',5,7-trihydroxydihydroflavone, CAS number 480-41-1, molecular formula C 15 h 12 o 5 , the molecular weight is 272.25. Naringenin is widely found in Rutaceae plants, such as grapefruit, tomato, grape and citrus fruits. Pharmacological studies have shown that naringenin has a wide range of physiological activities, including antibacterial, anti-inflammatory, antioxidant, anti-tumor, lowering blood fat, anti-aging, cough and expectorant, etc., so it can be developed and applied in the fields of medicine and food health care . [0003] The production of naringenin can be directly extracted from some natural plants, such as peach le...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P17/06C12R1/685
CPCC12P17/06C12N1/145C12R2001/685
Inventor 陈虹陈蔚青陆胤张建芬柯薇
Owner ZHEJIANG SHUREN UNIV
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