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Method for hydrolytic preparing biological tangeritin by enzyme

A technology for flavonoid monoglycoside and enzymatic preparation, which is applied in the field of enzymatic hydrolysis to prepare citrus bioflavonoid monoglycoside, can solve the problem of difficulty in obtaining naringenin monoglycoside or hesperetin monoglycoside, and achieves convenient separation and purification of products , The enzymatic hydrolysis process is easy and the effect of less by-products

Inactive Publication Date: 2007-12-19
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is also difficult to obtain naringenin monoglycoside or hesperetin monoglycoside by enzymatic hydrolysis of naringin or hesperidin
So far, the extraction of naringin and hesperidin has been relatively mature technology at home and abroad, but it has failed to provide an effective method to obtain naringin monoglycoside and hesperetin monoglycoside.

Method used

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  • Method for hydrolytic preparing biological tangeritin by enzyme
  • Method for hydrolytic preparing biological tangeritin by enzyme

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: preparation of solid enzyme

[0023] Aspergillus niger WZ001 (Zhejiang University of Technology Institute of Biology and Environmental Engineering provides) after 5 days of Cha's medium activation, inoculate a loop to the liquid seed medium (4% bran, 3% bean cake powder, 8% bean dregs, and the remainder is water) in 100 mL for 36 h to obtain a seed solution.

[0024] Liquid fermentation medium: 5% bran, 4% bean cake powder, 10% bean dregs, the remaining water, natural pH, 0.1Mpa steam pressure sterilization for 20 minutes, cooling, and adding Aspergillus niger seed liquid to each bottle at a volume ratio of 2%. Fermentation temperature 30-37 ℃, rotation speed 150-200r / min, cultured for 5-7 days, filtered to get the crude enzyme liquid, the enzyme activity of hesperidinase was 14104.9U / g fermentation raw material, the enzyme activity of naringinase The activity is 14799.6U / g fermentation material. Add 270mL of ethanol to 100mL of the obtained crude enzyme...

Embodiment 2

[0025] Embodiment 2: Preparation of immobilized enzyme

[0026] Aspergillus niger WZ001 was activated by Chase medium for 5 days, inoculated one loop into 100 mL of liquid seed medium (4% bran, 3% bean cake powder, 8% bean dregs, and the balance was water) and cultivated for 36 hours to obtain seed liquid.

[0027] Liquid fermentation medium: 5% bran, 4% bean cake powder, 10% bean dregs, the remaining water, natural pH, 0.1Mpa steam pressure sterilization for 20 minutes, cooling, and adding Aspergillus niger seed liquid to each bottle at a volume ratio of 2%. The fermentation temperature is 30-37° C., the rotation speed is 150-200 r / min, and the cultivation is carried out for 5-7 days, and the crude enzyme liquid is obtained by filtration.

[0028] Method (1): Take 10ml of enzyme solution, mix with 50mL of sodium alginate solution with a mass concentration of 1.5%, add 0.02g of tannic acid and 0.02g of polyvinyl alcohol, granulate with a syringe, and drop into 100mL of a mass ...

Embodiment 3

[0031] Embodiment 3: Preparation of naringenin monoglycoside

[0032] 0.2 g of naringin was dissolved in 100 ml of 20% dimethylformamide organic solution with a volume concentration of 20%, so that the concentration of naringin reached 2 g / L. Under the conditions of pH 3.0-6.0 and temperature 20-60°C, add 1-10g of glucose, use free or immobilized naringinase to enzymatically hydrolyze naringin, the enzymatic hydrolysis time is 20-60min, and quantify the product by HPLC Analysis, detection of naringenin monoglycoside and rhamnose content:

[0033]1. Under the conditions of pH 3.5 and temperature 20°C, add the solid enzyme 7U / mg naringin obtained in Example 1, add 1g glucose, and enzymatically hydrolyze it for 20 minutes to obtain 40 mg naringenin monoglycoside and 14.2 mg rhamnose, After enzymatic hydrolysis, the conversion liquid is separated with a macroporous adsorption resin, and eluted with a 30% volume concentration ethanol solution. The obtained substance is naringenin ...

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Abstract

In the invented method, citrus bioflavonoid and corresponding enzyme are used as raw materials, being dissolved in organic solvent, adding glucoside solution at concentration of 1-100g / L, under pH value of 3.0-7.0 and temperature of 20-65 deg.C for 20-60 min. After separation and purification, obtained is citrus bioflavonoid monoglycoside. With this invention, naringoside enzymolysis and cirmtim enzymolysis method for production of naringen in monoglycoside and hesperetin monoglycoside, compared with chemical method, has advantages of:easily to be controlled of enzymolysis, mild reaction, less by-products and easy to be separated of products. By using this invented method, the enzymolysis of naringoside and cirmtim for production of naringenin monocoside and hesperetin monoglycoside can be effectively controlled, and only the first glycosidic bond is broken, so obtained is highest yield of naringenin monoglycoside, hesperetin monoglycoside and rhamnose.

Description

(1) Technical field [0001] The invention relates to a method for preparing citrus bioflavonoid monoglycoside by enzymatic hydrolysis, in particular to a method for obtaining naringenin monoglycoside or hesperetin monoglycoside by enzymatically hydrolyzing naringin or hesperidin. (2) Background technology [0002] Most of the citrus bioflavonoids exist in the form of flavonoid glycosides, and the flavanone glycosides hesperidin (Hesperidin) and naringin are the most abundant in citrus fruits. The molecular structure of naringin (narirutin) is naringenin-7-β-neohesperidose, which is the main bitter substance of citrus and greatly affects the quality of citrus products. The naringin extracted from citrus has a strong bitter taste, and its application is limited. Hesperidin, also known as tangeridin, hesperidin, hesperetin 7-O-rutinoside, hesperetin-7-rhamnoglucoside, its chemical structure is a molecule of rutin and hesperetin (Hesperedin) composition Glycosides, hesperidin h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/06
Inventor 汪钊申丽静曹小燕沈雪亮章银军
Owner ZHEJIANG UNIV OF TECH
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