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Method for preparing carrageenin oligosaccharides by compositely degrading eucheuma with carrageenanase and cellulase

A technology of carrageenan oligosaccharides and cellulase, which is applied in fermentation and other directions, and can solve problems such as consumption, large energy resources, and uneven distribution of products

Inactive Publication Date: 2014-03-12
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The traditional preparation method of carrageenan oligosaccharides is to pulverize Eucheuma oligosaccharides, add a large amount of acid and acidify under high temperature conditions, but the reaction conditions of this method are not easy to control, the product distribution is uneven, and it is difficult to prepare a large amount of high-purity oligosaccharides
There are also people trying to use κ-carrageenanase to directly degrade carrageenan to prepare carrageenan oligosaccharides, but because the yield of carrageenase used is low, it is not suitable for large-scale production, and the extraction process of carrageenan consumes a large amount of energy resources. The sulfate group in the gum has a removal effect, so it is not conducive to the preparation of highly active carrageenan sulfated oligosaccharides

Method used

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  • Method for preparing carrageenin oligosaccharides by compositely degrading eucheuma with carrageenanase and cellulase
  • Method for preparing carrageenin oligosaccharides by compositely degrading eucheuma with carrageenanase and cellulase

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Embodiment 1

[0018] 1. Wash and soak 20g of dried Eucheuma, the weight after rehydration is about 200g, add water and homogenate to a final volume of 600mL.

[0019] 2. Adjust the homogenate to pH 5.0, add cellulase according to the amount of 21,000 U / g dry Eucheuma, and shake and react at 50°C for 2.5h.

[0020] 3. Cool the reaction liquid to about 35°C, add carrageenase according to the amount of 5 U / g dry Eucheuma, and shake the reaction at 35°C for 8 hours. Boil the enzymolysis solution to kill the enzyme, then centrifuge at 5000rpm for 10min to remove solid particles such as seaweed residue.

[0021] 4. Concentrate the supernatant to 300mL by rotary evaporation, stir and add 300mL ethanol, and centrifuge at 8000rpm for 15min to remove the undegraded seaweed components.

[0022] 5. Concentrate by rotary evaporation again to 50mL, add 250mL ethanol, at this time, a large amount of white sugar precipitates appear. Centrifuge at 5000rpm for 10min to obtain a large amount of white precip...

Embodiment 2

[0025] 1. Wash and soak 5g of dried Eucheuma, the weight after rehydration is about 40g, add water and homogenate to a final volume of 200mL.

[0026] 2. Adjust the homogenate to pH 5.0, add cellulase according to the amount of 18,000 U / g dry Eucheuma, and shake at 50°C for 2 hours.

[0027] 3. Cool the reaction liquid to about 35°C, add carrageenase according to the amount of 5 U / g dry Eucheuma, and shake the reaction at 35°C for 12 hours.

[0028] 4. Centrifuge the enzymatic solution at 5000 rpm for 10 minutes to remove solid particles such as seaweed residue.

[0029] 5. Concentrate the supernatant to 100mL by rotary evaporation, add 100mL ethanol after stirring, and centrifuge at 8000rpm for 15min to remove undegraded seaweed components.

[0030] 6. Concentrate the supernatant to 20mL by rotary steaming, add 100mL ethanol to precipitate and collect carrageenan oligosaccharides, and dry them to obtain tetrasaccharides, hexasaccharides, octasaccharides, decasaccharides, and...

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Abstract

The invention relates to a method for preparing carrageenin oligosaccharides by compositely degrading eucheuma with carrageenanase and cellulase. The method comprises the following steps: homogenating eucheuma, and degrading with cellulase for some time; degrading with kappa-carrageenanase under the following reaction conditions: the substrate concentration is 3-4%, the cellulase addition amount is 9-21 thousand U / g dry eucheuma, the cellulase degradation is performed at 45-55 DEG C under the pH value of 4.0-7.0 for 1-3 hours, the carrageenanase addition amount is 3-9 U / g dry eucheuma, and the carrageenanase degradation is performed at 35-45 DEG C under the pH value of 5.5-5.8 for 4-12 hours; and centrifuging to remove impurities, precipitating with 0.5-1.5 times of alcohol to remove degraded seaweed components, precipitating with 3-8 times of alcohol, centrifuging to collect the precipitate, and drying the precipitate to obtain the carrageenin oligosaccharides. The method for preparing the carrageenin oligosaccharides by directly compositely degrading eucheuma with carrageenanase and cellulase has the advantages of mild reaction conditions, single product, high yield of active oligosaccharides, no pollution, low energy consumption, short time consumption, low production cost and the like, and can be used for mass production of carrageenin oligosaccharides of which the molecular weight is 600-2000Da.

Description

technical field [0001] The invention relates to a method for preparing carrageenan oligosaccharides by compounding carrageenanase and cellulase to degrade Eucheuma. Background technique [0002] Eucheuma, a marine red algae, is the main raw material source of carrageenan. Among them, the main components of Kappa auris, Kappa heterocladum, and Kappa longicardium are κ-carrageenan, which is an important raw material for the industrial production of κ-carrageenan, and its polysaccharide is a natural sulfated polysaccharide, which has High sulfate content. κ-carrageenan is composed of 3,6-endether-α-D-galactose and 4-sulfate-β-D-galactose linked by β-1,4-glucosidic bonds alternately. In recent years, polysaccharides, as a new type of antiviral drug, have received great attention from scientific researchers, and in the field of polysaccharides, sulfated polysaccharides are a research hotspot. It has been reported that carrageenan oligosaccharides and their derivatives have goo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/14
Inventor 牟海津刘哲民段芳媛田琳
Owner OCEAN UNIV OF CHINA
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