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Method for preparing series of neoagarool igosaccharode with single degree of polymerization by utilizing immobilized enzyme

A new agar oligosaccharide, immobilized enzyme technology, applied in biochemical equipment and methods, microorganism-based methods, immobilized on or in inorganic carriers, etc., can solve the problem that the activity is easily affected by enzymatic hydrolysis conditions, product separation Purification effect, difficulty in rapid development and other problems, to achieve the effect of simple and effective immobilization method, reuse, and simplify post-processing steps

Pending Publication Date: 2019-03-15
HUAQIAO UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Although the enzymatic hydrolysis method has many advantages, the activity of agarase, the key catalyst of the enzymatic hydrolysis reaction, is easily affected by the enzymatic hydrolysis conditions and is difficult to reuse. At the same time, it will also have a certain impact on the separation and purification of the product.
These deficiencies are also one of the reasons why the industrial development of new agar oligosaccharides is difficult to develop rapidly. Therefore, how to improve the activity, stability and reuse of agarase and make it an industrial catalyst has become one of the problems that need to be broken through in current research.

Method used

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  • Method for preparing series of neoagarool igosaccharode with single degree of polymerization by utilizing immobilized enzyme
  • Method for preparing series of neoagarool igosaccharode with single degree of polymerization by utilizing immobilized enzyme
  • Method for preparing series of neoagarool igosaccharode with single degree of polymerization by utilizing immobilized enzyme

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Experimental program
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Effect test

Embodiment 1

[0025] The preparation of embodiment 1 agarase enzyme liquid

[0026] Vibrio natriegens HJPHYXJ-1 (disclosed in patent application 201510390422.9, agarase-producing vibrio natriegens and its application, publication number CN105087427A, is preserved in China Type Culture Collection Center, address: Wuhan, China , Wuhan University, postal code: 430072, preservation number: CCTCC M2015244, preservation date: April 23, 2015) transferred from the slant culture medium to the seed culture medium, obtained the seed liquid by shaking culture, and inoculated the seed liquid into the liquid medium In this method, the inoculum size is 6%, the fermentation temperature is 35°C, the stirring speed is 100rpm, and the fermentation time is 48h. The supernatant obtained after the fermentation broth is centrifuged to remove bacteria is separated by ammonium sulfate precipitation, dialyzed, and finally freeze-dried to obtain agarase powder.

[0027] See CN105087427A for the specific formulations ...

Embodiment 2

[0028] The preparation of embodiment 2 immobilized enzyme

[0029] (1) Take 1.0g of graphene oxide (GO) and add it to 80mL of 60% ethanol solution. After ultrasonic dispersion for 0.5h, add 0.676g of ferric chloride hexahydrate and 0.695g of ferrous sulfate heptahydrate, and continue ultrasonication for 20min. Add 3.5mol L ～1 After 100 mL of ammonia solution was reacted at reflux temperature of 60 °C for 2 h, the product was separated with a magnet, washed with deionized water until neutral, and dried to obtain GO / Fe 3 o 4 Composite magnetic carrier.

[0030] The above-mentioned ferric chloride hexahydrate can also be replaced with at least one of other ferric chloride or its hydrate, ferric sulfate or its hydrate and iron nitrate or its hydrate of the same molar amount, and the above-mentioned ferrous sulfate heptahydrate can also be It can be replaced with at least one of other ferrous sulfate heptahydrate or its hydrates, ferrous chloride or its hydrates, and ferrous nit...

Embodiment 3

[0032] The influence of embodiment 3 glutaraldehyde concentration on immobilized agarase enzymatic activity

[0033] Agarase and immobilized carrier preparation and immobilization method refer to Examples 1 and 2, the difference is that when immobilizing the enzyme, the volume concentration of glutaraldehyde aqueous solution is 1%, 2%, 3%, 4%, 5%.

[0034] Table 1 Effect of glutaraldehyde concentration on immobilized agarase activity

[0035] Glutaraldehyde concentration (%)

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Abstract

The invention discloses a method for preparing a series of neoagarool igosaccharode with single degree of polymerization by utilizing an immobilized enzyme. The method comprises the following steps: (1) acquiring agarase powder from vibrio natriegens HJPHYXJ-1; (2) preparing an immobilized carrier; (3) preparing the immobilized enzyme; (4) preparing a series of neoagarool igosaccharode. Accordingto the method, an immobilized agarase is prepared by adopting an immobilized enzyme technology and is applied in the preparation of the neoagarool igosaccharode; through simple magnetic separation, reutilization of the agarase can be realized, and aftertreatment steps are simplified, therefore, the production cost of the neoagarool igosaccharode is greatly lowered; additionally, the immobilized carrier used in the method is simple to prepare, and an immobilization method is simple and effective, so that the theoretical and technical supports can be provided for future industrial large-scale production of the neoagarool igosaccharode.

Description

technical field [0001] The invention belongs to the technical field of preparation of new agar oligosaccharides, and in particular relates to a method for preparing a series of new agar oligosaccharides with a single degree of polymerization by using immobilized enzymes. Background technique [0002] Agarose is mainly derived from the cell walls of red algae such as Gracilaria, Geliflower, and Porphyra, and is linked by β-D-galactofuranose linked by α(1→3) glycosidic bonds and β(1→4) glycosidic bonds. A linear neutral polysaccharide formed by alternating linkages of 3,6-endether-α-L-galactofuranose. The new agar oligosaccharide is an oligosaccharide with a polymerization degree of 2-10, which is obtained by hydrolysis of agarose and is connected by agarose as a repeating unit, with D-galactose as the reducing end. The new agar oligosaccharide has a variety of physiological activities, including hypoglycemic, hypolipidemic, anti-inflammatory, anti-oxidant, prebiotic, whiteni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14C12P19/04C12N11/14C12R1/63
CPCC12N11/14C12P19/04C12P19/14
Inventor 叶静张娜肖美添林福娣黄雅燕张学勤
Owner HUAQIAO UNIVERSITY
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