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Production method of hesperidinase and application

A technology of hesperidinase and production method, which is applied in the field of microbial fermentation and can solve the problems of poor thermal stability and low enzyme activity of the enzyme

Active Publication Date: 2016-05-25
成都康辉生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a production method of hesperidinase with mild reaction conditions, good selectivity and simple and optimized process, so as to solve the problems of low enzyme activity and poor thermal stability of the existing hesperidinase

Method used

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  • Production method of hesperidinase and application
  • Production method of hesperidinase and application
  • Production method of hesperidinase and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] A production method of hesperidinase, comprising the following steps:

[0098] 1), Aspergillus niger is carried out sporulation culture in substratum to form Aspergillus niger spore;

[0099] The culture medium is composed of the following components in parts by weight: 100 g of wheat bran, 70 ml of distilled water;

[0100] Described sporulation culture refers to that 1ml spore suspension (concentration is 10 6 individual / ml) Aspergillus niger spores were inoculated into 50g bran medium, and cultivated for 6 days at 30°C; until the bran medium turned dark brown completely, it indicated that Aspergillus niger had formed a large number of spores, and then used 500ml aseptic water to elute the spores to a spore concentration of 10 6 pcs / ml spare;

[0101] 2), Aspergillus niger spores are fermented in the enzyme-producing medium to produce hesperidinase;

[0102] The enzyme-producing medium is composed of the following components by weight percentage: 0.2% NaNO 3 , 0....

Embodiment 2

[0111] A production method of hesperidinase, comprising the following steps:

[0112] 1), Aspergillus niger is carried out sporulation culture in substratum to form Aspergillus niger spore;

[0113] The culture medium is composed of the following components by weight: 100g of wheat bran, 70ml of water;

[0114] Described sporulation culture refers to that 1ml spore suspension (concentration is 10 6 individual / ml) Aspergillus niger spores were inoculated into 50g bran medium, and cultivated for 7 days at 35°C; until the bran medium turned dark brown completely, it indicated that Aspergillus niger had formed a large number of spores, and then used 1000ml aseptic water to elute the spores to a spore concentration of 10 6 pcs / ml spare;

[0115] 2), Aspergillus niger spores are fermented in the enzyme-producing medium to produce hesperidinase;

[0116] The enzyme-producing medium is composed of the following components by weight percentage: 0.2% NaNO 3 , 0.005% KCl, 0.1% K 2 ...

Embodiment 3

[0125] A production method of hesperidinase, comprising the following steps:

[0126] 1), Aspergillus niger is carried out sporulation culture in substratum to form Aspergillus niger spore;

[0127] The culture medium is composed of the following components in parts by weight: 100 g of wheat bran, 70 ml of distilled water;

[0128] Described sporulation culture refers to that 1ml spore suspension (concentration is 10 6 Individual / ml) Aspergillus niger spores were inoculated into 50g bran medium, and cultivated for 6 days at 33°C; until the bran medium turned dark brown completely, it indicated that Aspergillus niger had formed a large number of spores, and then used 800ml aseptic water to elute the spores to a spore concentration of 10 6 pcs / ml spare;

[0129] 2), Aspergillus niger spores are fermented in the enzyme-producing medium to produce hesperidinase;

[0130] The enzyme-producing medium is composed of the following components by weight percentage: 0.2% NaNO 3 , 0....

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Abstract

The invention discloses a production method of hesperidinase and an application. The production method comprises the following steps: (1) carrying out spore-producing culture on aspergillus niger in a bran culture medium to form aspergillus niger spores; (2) carrying out fermentative enzyme production on the aspergillus niger spores in an enzyme-producing culture medium to produce the hesperidinase; and (3) carrying out separation and extraction to obtain an enzyme liquid, wherein the aspergillus niger is preserved in the China General Microbiological Culture Collection Center; the preservation name is KH005; and the preservation number is CGMCC12101. The hesperidinase produced by the aspergillus niger KH005 in an enzyme-production and fermentation culture medium has the advantages of high enzyme activity and good thermal stability. The aspergillus niger KH005 is applied to production of hesperetin by an enzymolysis approach; a new process selection is provided for production of the hesperetin; the enzymolysis approach is mild in reaction condition, good in selectivity; a parent ring of the hesperetin is not easy to denature; and the produced hesperetin is high in content and easy to separate.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a production method and application of hesperidinase. Background technique [0002] Hesperidinase is the key enzyme for hydrolyzing hesperidin and has high application value. Hesperetin monoglycoside, hesperetin, rhamnose and glucose can be obtained by hydrolyzing hesperidin with hesperidinase. In addition, hesperidinase is added to canned orange petals to hydrolyze the hesperidin in it, which can prevent white precipitation in canned oranges. [0003] Hesperidinase can be produced by a variety of microorganisms, such as Penicillium, Aspergillus oryzae, Aspergillus niger and Bacteroides. At present, domestic research on hesperidase-producing strains is still relatively small, and most of the reported hesperidase-producing strains are molds. [0004] In the existing method of producing hesperidinase by using mold, the production cost of hesperidinase is relatively high due...

Claims

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Application Information

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IPC IPC(8): C12N9/24C12P17/06C12R1/685
CPCC12N9/24C12P17/06C12Y302/0104
Inventor 陈文清魏勇冯唯成
Owner 成都康辉生物科技有限公司
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