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Low-viscosity fucosan and high-temperature high-pressure enzymatic coupling preparation method thereof

A technology of viscosity fucoidan, high temperature and high pressure, applied in lyase, fermentation, etc., can solve the problems of low sulfate retention and low enzymatic hydrolysis efficiency, and achieve the effect of low cost, high hydrolysis efficiency and mild reaction conditions

Active Publication Date: 2019-09-24
海南原海高慧科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome the defects of low sulfate retention in the existing acid, alkali degradation and oxidation degradation of fucoidan and low enzymolysis efficiency in the existing biological enzymatic degradation of fucoidan , and provide a method for preparing low-viscosity fucoidan with high temperature and high pressure-enzyme coupling coupling with high sulfate retention and high enzymolysis efficiency, and the low-viscosity fucoidan prepared by the method

Method used

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  • Low-viscosity fucosan and high-temperature high-pressure enzymatic coupling preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Fucoidan prehydrolysis process optimization

[0036] In this example, by comparing the hydrolysis effect of fucoidan by acid method, oxidation method and high temperature and high pressure treatment technology, the best pre-hydrolysis method is selected based on the retention of sulfuric acid group when the viscosity value of the polysaccharide solution drops by about 20%. process conditions.

[0037] (1) Experimental method

[0038] A. Acid hydrolysis: Prepare 2wt% aqueous solution of high-polymeric fucoidan, and add HCl solution to make the final concentrations 0, 0.05mol / L, 0.1mol / L and 0.2mol / L respectively, and then Hydrolysis was carried out with or without ultrasonic (frequency 40kHz) auxiliary technology, and the change of viscosity value during the hydrolysis process was measured every 15 minutes. After the reaction, the pH value of the hydrolyzate was adjusted to neutrality, and then passed through an 8kDa ultrafiltration membrane to remove the fa...

Embodiment 2

[0045] Example 2: Hydrolysis effect of fucoidan lyase on pretreated fucoidan

[0046] In this example, the fucoidan lyase extracted from scallop viscera or abalone viscera was used as a tool enzyme to further hydrolyze the high-temperature and high-pressure pre-hydrolyzed fucoidan solution obtained in Example 1, and the viscosity value of the polysaccharide solution was changed. Observe the hydrolysis effect as an indicator.

[0047] (1) Experimental method

[0048] A. Preparation of fucoidan lyase: After the fresh scallop viscera or abalone viscera are broken and homogenized by a homogenizer, the resulting homogenate is added with pH=7.0 at a ratio of 1:5 (g / mL). Phosphate buffer solution, leaching overnight at 4°C; refrigerated centrifugation (8000r / min, 20min), collecting the supernatant; adding ammonium sulfate to the supernatant to make the ammonium sulfate concentration 30% (w / v), And leave it to react at 4°C for 2h, refrigerate and centrifuge (8000r / min, 15min), colle...

Embodiment 3

[0054] Embodiment 3: the formula design of compound hydrolase

[0055] (1) Experimental method

[0056] A. The preparation of compound hydrolase: the fucoidan lyase that embodiment 2 obtains and β-galactosidase (30000U / g), β-glucosidase (30000U / g), alkaline protease (20000U / g) g) and pectinase (8000U / g) are compounded according to the weight ratio of 6:1:1:1:1 to form a composite hydrolase.

[0057] B. Hydrolysis effect: Dissolve high-polymeric fucoidan in deionized water to obtain a 2wt% aqueous solution of high-polymeric fucoidan; use the preferred high-temperature and high-pressure pretreatment conditions of Example 1 (110°C / 0.08MPa, 2-5 minutes) to pretreat the high-polymer fucoidan aqueous solution; after cooling to room temperature, add complex hydrolase to the pretreated fucoidan solution so that the final concentration is 0.01-0.05wt%, at room temperature After hydrolysis for 1-6 hours, analyze the viscosity change of the fucoidan solution.

[0058] C. Drying: spray...

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Abstract

The invention belongs to the field of biological preparations, and discloses a low-viscosity fucosan and a high-temperature high-pressure enzymatic coupling preparation method thereof. The method includes the following steps that 1, a high polymer fucosan aqueous solution is subjected to high-temperature high-pressure treatment, and a pre-hydrolysis solution is obtained; 2, a phosphate buffered solution is added into scallop viscera and / or abalone viscera obtained after tissue homogenization, the mixture is subjected to extraction and refrigerated centrifugation, supernate is collected, ammonium sulfate is added into the supernate, the mixed liquid is subjected to standing for reaction, freeze centrifugation is conducted, supernate is collected, ammonium sulfate is added into the supernate, then after a precipitate is dissolved, ultrafiltration membrane filtration and freeze-drying are conducted sequentially, and fucosan lyase is obtained; the fucosan lyase is mixed with beta-galactosidase, beta-glucosidase, alkaline protease and pectinase to obtain compound hydrolases; a pre-hydrolysis solution is subjected to enzymolysis under the action of compound hydrolases. The provided method is adopted for preparing the low-viscosity fucosan, the enzymolysis efficiency is high, the sulfuric acid group retention degree is not less than 90%, and the recovery rate is not less than 90%.

Description

technical field [0001] The invention belongs to the field of biological preparations, and in particular relates to a low-viscosity fucoidan and a high-temperature, high-pressure-enzyme coupling preparation method thereof. Background technique [0002] Fucoidan exists in the cell wall matrix of brown algae (such as Ascophyllum Nodosum, Fccus vesiculosus, Laminaria japonica), and is a protective mucopolysaccharide in the intertidal zone of brown algae. Fucoidans from different sources are quite different in structure such as monosaccharide composition, sulfate content and position, molecular weight, connection mode, and glycosyl connection sequence. According to reports, fucoidan has a variety of biological activities, such as anti-inflammatory, anti-virus, anti-coagulation, anti-tumor, etc., and is safe and has no toxic side effects. However, natural fucoidan has high degree of polymerization, large molecular weight (≥189kDa), and low biological activity, all of which hinder...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/88C12P19/14C12P19/04
CPCC12N9/88C12P19/04C12P19/14
Inventor 刘翼翔邹超傅秋叶张亚平陈昭华
Owner 海南原海高慧科技有限公司
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