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Method for preparing hesperetin monoglucoside by fermentation, enzymolysis, extraction and coupling

A technology of monoglucoside and hesperetin, which is applied in the field of conversion and extraction of biochemical substances, can solve the problems of high price, easy degradation, and low yield of hesperidinase, and achieve reduced production costs, improved conversion rates, and improved product quality. yield effect

Active Publication Date: 2013-02-13
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The object of the present invention is to propose a method for preparing hesperetin monoglucoside by enzymatically hydrolyzing hesperidin with hesperidinase, which has low yield, easy to be degraded, and high price of hesperidinase. The method has a high yield and can not only reduce the production cost, but also prevent the generated hesperetin monoglucoside from being further degraded into hesperetin by coupling fermentation-enzymolysis-extraction to prepare hesperetin monoglucoside

Method used

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  • Method for preparing hesperetin monoglucoside by fermentation, enzymolysis, extraction and coupling

Examples

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Embodiment 1

[0046] A method for preparing hesperetin monoglucoside by coupling fermentation-enzymolysis-extraction, the method is carried out according to the following steps:

[0047] 1) Preparation of culture medium: the components of the following two types of culture medium and the mass percentage of each component are as follows:

[0048] a. Bacteria activation culture medium: 3% glucose, 0.3% beef extract, K 2 HPO 4 ·3H 2 O 0.3%, MgSO 4 ·7H 2 O 0.05%, FeSO 4 ·7H 2 O 0.01%, make up to 100% with water, and reserve;

[0049] b. Production of fermentation broth: 3% sucrose, 0.3% beef extract, K 2 HPO 4 ·3H 2 O 0.3%, MgSO 4 ·7H 2 O 0.05%, FeSO 4 ·7H 2 O 0.01%, make up to 100% with water, and reserve;

[0050] 2) Preparation of strain seed solution: Fusarium nivale ( Fusarium nivale ) HESPER strain CGMCC No.6238 is the strain, pick a small amount of strains in the strain activation culture medium, activate and cultivate at 30 ℃, 150 r / min for 48 h, until the number ...

Embodiment 2

[0058] In this example, step 2) preparation of strain seed liquid: culture speed is 170 r / min, activation time is 36 h; step 3) preparation of fermentation liquid for hesperidin production: strain seed liquid: production fermentation culture liquid: Hesperidin = 1.5mL: 75mL: 0.75g; Step 4) Cultivation of Hesperidin Production Fermentation Broth: Sterile air input rate is 2.5 L / min, stirring speed is 50 r / min; Step 5) Resin of Fermentation Broth Adsorption, reflux, discharge and feeding: the macroporous resin adopts the AB-8 model, the reflux rate of the fermentation broth is 2 column volumes / h, the material discharge interval is 20 h, the material discharge is 20%, and the feed is 20% at the same time ; Step 6) Elution and collection of resin: the amount of water washing is 5 column volumes / h, the ethanol concentration is 85%, and the ethanol elution rate is 1 column volume / h; the remaining steps and processes are the same as in Example 1.

[0059]

Embodiment 3

[0061] In this example, step 2) preparation of strain seed liquid: culture speed is 180 r / min, activation time is 24 h; step 3) preparation of fermentation liquid for hesperidin production: strain seed liquid: production fermentation culture liquid: Hesperidin = 2mL: 100mL: 1g; Step 4) Cultivation of Hesperidin Production Fermentation Broth: Input rate of sterile air was 5 L / min, stirring speed was 80 r / min; Step 5) Resin adsorption of Fermentation Broth, Reflux, discharge and feeding: the macroporous resin adopts the D101-I model, the reflux rate of the fermentation broth is 4 column volumes / h, the material discharge interval is 30 h, the material discharge is 30%, and the feed is 30% at the same time; steps 6) Resin elution and collection: the amount of water washing is 8 column volumes / h, the ethanol concentration is 100%, and the ethanol elution rate is 2 column volumes / h; the rest of the steps and processes are the same as in Example 1.

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Abstract

The invention discloses a method for preparing hesperetin monoglucoside by fermentation, enzymolysis, extraction and coupling, belonging to the technical field of biochemical substances extraction. The method comprises the following steps: preparing a nutrient solution; preparing a strain seed solution; preparing fermentation liquor for producing the hesperetin monoglucoside; culturing the fermentation liquor for producing the hesperetin monoglucoside; carrying out resin adsorption, back flow, discharging and feeding on the fermentation liquor; and eluting and collecting resin. According to the method disclosed by the invention, the Fusarium nivale HESPER strain CGMCCNo.6238 is adopted as a culture, and the hesperetin monoglucoside is prepared by continuous culture fermentation, hesperidin enzymolysis, extraction and coupling, thus the hesperetin monoglucoside is effectively prevented from being degraded into hesperetin, the yield is enhanced, and the production cost is reduced. The method disclosed by the invention can be popularized and applied to related areas and enterprises.

Description

technical field [0001] The invention relates to the technical field of conversion and extraction of biochemical substances, in particular to a method for preparing hesperetin monoglucoside by coupling fermentation-enzymolysis-extraction. Background technique [0002] Citrus is the world's largest fruit, and my country's citrus planting area and output both rank first in the world. Hesperidin is the main flavonoid in citrus. Due to the poor water solubility and absorption of hesperidin, the pharmacological effect is relatively weak. Remove a molecule of rhamnoside from hesperidin by chemical or enzymatic methods to generate hesperetin monoglucoside, which can improve its water solubility and effectively enhance its efficacy. The current market price of hesperetin monoglucoside is as high as 1,000 yuan / More than kg. However, the hydrolysis reaction conditions of the chemical method are difficult to grasp. At present, it has been reported that hesperidin monoglucoside is ge...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12P19/60C12R1/77
Inventor 张俊夏其乐陈剑兵邢建荣陆胜民李小莉
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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