Method for preparing purified Naringoside acid, and application of enzyme preparation

A technology for purifying naringinase and naringinase, which is applied in the directions of hydrolase, fungi, food science, etc., can solve problems such as failure to form industrialized production, and achieve broad-spectrum substrate hydrolysis activity and temperature, high conversion rate, Ease of cultivation and reproduction

Inactive Publication Date: 2007-12-26
颜廷和
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0003] At present, the research on naringinase in China is in the stage of bacterial strain screening, and industrial production has not been formed. However, through the isolation of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The preparation of naringinase is operated as follows: 1) take Aspergillus fungus as bacterial classification, named aspergillus niger, morphological feature is: bacterial strain is on Cha's culture medium, and aerial hyphae grows well, and cotton-wool shape has protrusion, surface White, the color of the back turns from light brown to brown after prolonged time, and the spores are brown; adopt conventional microbial inoculation method to inoculate: culture on the slant of test tube, adopt Chapei solid medium; transfer (enzyme production): transfer the above-mentioned grown strains Into the sporulation solid medium, 28 ℃, cultivated for 3 days, until the spores are produced: the sporulation medium uses 1000ml of 20 potato leaching juice, 10g of glucose, 3g of potassium dihydrogen phosphate, 1.5g of magnesium sulfate, and a trace amount of vitamin B1. Agar 20g, put a filter paper strip in each inclined test tube; 2) above-mentioned bacterial strain spore is added to solid...

Embodiment 2

[0021] The difference from Example 1 is:

[0022] In the solid fermentation step 2) of naringinase, the spores of the belonging strains are added to the solid culture concentration by 3% inoculum, the carbon in the medium is wheat bran, the nitrogen is bean cake and bean cake powder, and the control carbon: the ratio of nitrogen is 2:1, keep humidity 30%, temperature 25°C, pH is natural value, naringinase is produced in 60 hours; the naringinase culture produced is soaked with water, filtered, and the filtrate is used as a debittering agent for orange juice, and the addition amount is 0.1‰-2‰ of orange juice.

[0023] The naringinase prepared by the invention is used as the debittering agent of orange juice, and the weight gain effect is remarkable.

Embodiment 3

[0025] The difference from Example 1 is that in the naringinase fermentation preparation step 2), the spores of the strain are added to the solid medium according to 5% inoculum size, and the carbon in the solid medium is wheat bran, and the nitrogen is soybean cake and bean cake powder, control the ratio of carbon and nitrogen to 5:4, keep the humidity at 70%, temperature at 32°C, pH at natural value, naringinase will be produced in 90 hours; soak the produced naringinase in water, add ammonium sulfate to precipitate, control The saturation is 60%, and it is made into a pure enzyme preparation, and as a debittering agent for orange juice, the added amount is a few parts per million of the orange juice.

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Abstract

This invention discloses a method for preparing and purifying naringinase. The method comprises: (1) culturing bacterial strain with Czapek's medium, inoculating in a sporification medium, and culturing for 3-5 days to obtain spores as seeds; (2) inoculating the seeds (0.5-10%) in a solid medium, controlling C/N ratio at (2-5):(1-3.5), culturing at 25-35 deg.C, 30-70% humidity, and natural pH value for 40-90 h, extracting, adding 2.5 times pure water, centrifuging, filtering the supernatant, and precipitating with ammonium sulfate (saturation degree = 30-80%) and 40-90% organic solvent in sequence. The naringinase product has high activity, and can be used to remove bitterness of orange, and convert flavonoids aglycone in natural medicines.

Description

(1) Technical field [0001] The invention relates to production, preparation and purification of naringinase and its application in debittering of citrus products and biotransformation of flavonoids, in particular to a preparation and purification method of naringinase and its application in food and medicine. (2) Background technology [0002] Naringinase is a debittering enzyme produced by fungi. As an efficient biocatalyst, naringinase is also called a terminal non-reducing β-rhamnose residue decomposing enzyme in the β-rhamnoside molecule, releasing rhamnose. Naringinase can be produced by Aspergillus niger, Aspergillus oryzae and Penicillium. According to reports at home and abroad, the enzyme can be used for debittering of orange juice, canned oranges, and orange peels. [0003] At present, the research on naringinase in China is in the stage of strain screening, which has failed to form industrialized production. However, through the isolation of strains and continuou...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12N1/14A23L2/66
Inventor 颜廷和
Owner 颜廷和
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