Extraction method of algal polysaccharides

Abstract

The invention discloses an extraction method of algal polysaccharides, which comprises the following steps: putting seaweed powder, neutral cellulase, pectinase and papain into water for enzymolysis, and then performing high-temperature and high-pressure extraction and filtration to obtain an algal polysaccharide crude extracting solution; carrying out alcohol precipitation, centrifugation and absolute ethyl alcohol washing on the crude algal polysaccharide extracting solution to obtain a crude algal polysaccharide extract; dissolving the algal polysaccharide crude extract in water for purification treatment to obtain the algal polysaccharide, according to the extraction method of the algal polysaccharides, the raw materials are smashed firstly, extraction of the algal polysaccharides in the raw materials is facilitated, enzymolysis is conducted through cellulase, pectinase and papain, macromolecular cellulose and protein in the raw materials can be decomposed, sufficient precipitation of the algal polysaccharides is promoted, and finally the algal polysaccharides are extracted. The extraction of the algal polysaccharides can be fully realized by the assistance of high-temperature and high-pressure extraction, the extraction rate of the algal polysaccharides is remarkably improved, and meanwhile, the purity of the prepared algal polysaccharides is improved through a series of purification processes.

Description

Technical Field

[0001] The invention relates to the technical field of seaweed polysaccharide extraction, and in particular to a method for extracting seaweed polysaccharide. Background Art

[0002] There are two types of bioactive substances in seaweed. One is smaller in molecular weight and can directly or indirectly affect metabolites in the body after absorption, mainly including halogen compounds, terpenoid compounds, bromophenol compounds, hydroquinone, seaweed tannins, laminin, etc. The other is intercellular viscous polysaccharides that are difficult to digest and absorb, mainly including alginic acid, fucoidan, sulfated polysaccharides in brown algae, agar and carrageenan in red algae. Polysaccharides are important components of all living organisms and play an important role in controlling cell division, regulating cell growth and maintaining normal metabolism of living organisms. Seaweed polysaccharides mainly come from seaweeds such as kelp, Sargassum, giant kelp, Ascophyllum, and Fucus. Seaweed polysaccharides mainly include alginate, fucoidan and brown algae starch. The crude products of alginate, fucoidan and brown algae starch isolated from kelp are all white powders with a slight yellowish color.

[0003] At present, the extraction of seaweed polysaccharides is generally carried out through hot water extraction. Although this process can extract seaweed polysaccharides from algae, the extraction rate is low and the purity is low. It is impossible to fully extract seaweed polysaccharides from algae, resulting in a waste of resources.

[0004] In view of this, the present invention is proposed. Summary of the invention

[0005] The purpose of the present invention is to provide a method for extracting seaweed polysaccharides. The extraction method of the present invention can fully extract seaweed polysaccharides from algae and remove impurities through the control of various processes and parameters, thereby improving the extraction rate and purity of seaweed polysaccharides.

[0006] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are particularly adopted:

[0007] The present invention provides a method for extracting seaweed polysaccharides, which comprises the following steps:

[0008] (a) washing, drying and crushing seaweed to obtain seaweed powder;

[0009] (b) placing seaweed powder and neutral cellulase, pectinase and papain in water for enzymolysis, then extracting under high temperature and high pressure, filtering, and obtaining a crude seaweed polysaccharide extract;

[0010] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation, centrifugation, and washing with anhydrous ethanol to obtain a crude seaweed polysaccharide extract;

[0011] (d) dissolving the crude seaweed polysaccharide extract in water, and then sequentially performing decolorization, deproteinization and desalination treatments, and then performing alcohol precipitation and drying to obtain the seaweed polysaccharide.

[0012] Preferably, the drying is performed until the moisture content of the seaweed is 8% to 10%.

[0013] Preferably, the pulverization is performed to a particle size of not less than 80 meshes.

[0014] Preferably, the mass ratio of the seaweed powder to water is 1:(20-30).

[0015] Preferably, the addition amount of the neutral cellulase is 0.8% to 1.2% of the mass of the seaweed powder; the addition amount of the pectinase is 0.2% to 0.5% of the mass of the seaweed powder.

[0016] Preferably, the enzyme activity of the neutral cellulase is not less than 100,000 U / g; and the enzyme activity of the pectinase is not less than 20,000 U / g.

[0017] Preferably, the added amount of papain is 0.5% to 1% of the mass of the seaweed powder.

[0018] Preferably, the enzymatic activity of the papain is not less than 100,000 U / g.

[0019] The present invention does not specifically limit the decolorization, deproteinization and desalination methods, and conventional methods in the art may be used. Preferably, the decolorization method is an activated carbon adsorption method; the deproteinization method is a Sevage method; and the desalination method is an ion dialysis method.

[0020] Preferably, the enzymatic hydrolysis temperature is 50-55°C and the time is 1-2h.

[0021] Preferably, the high temperature and high pressure extraction pressure is 200-300 MPa, the temperature is 80-90° C., and the time is 20-40 min.

[0022] Compared with the prior art, the beneficial effects of the present invention include at least:

[0023] The seaweed polysaccharide extraction method of the present invention first crushes the raw material, which is beneficial to the extraction of seaweed polysaccharides in the raw material. At the same time, cellulase, pectinase and papain are used for enzymatic hydrolysis, which can decompose the macromolecular cellulose and protein in the raw material and promote the full release of seaweed polysaccharides. Finally, high temperature and high pressure extraction can fully realize the extraction of seaweed polysaccharides, significantly improve the extraction rate of seaweed polysaccharides, and at the same time, through a series of purification processes, the purity of the prepared seaweed polysaccharides is improved. DETAILED DESCRIPTION

[0024] The following embodiments of the technical solution of the present invention are described in detail in conjunction with the embodiments. The following embodiments are only used to more clearly illustrate the technical solution of the present invention, and are therefore only used as examples, and cannot be used to limit the scope of protection of the present invention.

[0025] It should be noted that, unless otherwise specified, the technical terms or scientific terms used in this application should have the common meanings understood by those skilled in the art to which the present invention belongs.

[0026] The raw materials used in the following examples are as follows:

[0027] Neutral cellulase: 100,000 enzyme activities, commercially available;

[0028] Papain: 100,000 enzyme activities, commercially available;

[0029] Pectinase: 26,000 enzyme activities, commercially available;

[0030] Bromelain: 100,000 enzyme activities, commercially available.

[0031] Example 1

[0032] This embodiment is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0033] (a) washing and drying the seaweed until the water content of the seaweed is 8.2%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0034] (b) placing seaweed powder, neutral cellulase, pectinase and papain in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction, filtering, and obtaining a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:20; the amount of neutral cellulase added is 0.8% of the mass of seaweed powder; the amount of pectinase added is 0.2% of the mass of seaweed powder; the amount of papain added is 1% of the mass of seaweed powder; the enzymolysis temperature is 50° C., the time is 2 h; the high temperature and high pressure extraction pressure is 200 MPa, the temperature is 90° C., and the time is 20 min;

[0035] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0036] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0037] Example 2

[0038] This embodiment is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0039] (a) washing and drying the seaweed until the water content of the seaweed is 9.6%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0040] (b) placing seaweed powder, neutral cellulase, pectinase and papain in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction, filtering, and obtaining a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:30; the amount of neutral cellulase added is 1.2% of the mass of seaweed powder; the amount of pectinase added is 0.5% of the mass of seaweed powder; the amount of papain added is 0.5% of the mass of seaweed powder; the enzymolysis temperature is 55° C., the time is 1 hour; the high temperature and high pressure extraction pressure is 300 MPa, the temperature is 80° C., and the time is 40 minutes;

[0041] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0042] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0043] Example 3

[0044] This embodiment is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0045] (a) washing and drying the seaweed until the water content of the seaweed is 8.8%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0046] (b) placing seaweed powder, neutral cellulase, pectinase and papain in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction and filtering to obtain a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:25; the amount of neutral cellulase added is 1.2% of the mass of seaweed powder; the amount of pectinase added is 0.3% of the mass of seaweed powder; the amount of papain added is 0.8% of the mass of seaweed powder; the enzymolysis temperature is 52° C., the time is 1.5 h; the high temperature and high pressure extraction pressure is 240 MPa, the temperature is 85° C., and the time is 35 min;

[0047] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0048] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0049] Example 4

[0050] This embodiment is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0051] (a) washing and drying the seaweed until the water content of the seaweed is 9.3%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0052] (b) placing seaweed powder, neutral cellulase, pectinase and papain in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction, filtering, and obtaining a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:28; the amount of neutral cellulase added is 0.8% of the mass of seaweed powder; the amount of pectinase added is 0.4% of the mass of seaweed powder; the amount of papain added is 0.8% of the mass of seaweed powder; the enzymolysis temperature is 53° C., the time is 2 h; the high temperature and high pressure extraction pressure is 280 MPa, the temperature is 85° C., and the time is 30 min;

[0053] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0054] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0055] Example 5

[0056] This embodiment is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0057] (a) washing and drying the seaweed until the water content of the seaweed is 9.4%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0058] (b) placing seaweed powder, neutral cellulase, pectinase and papain in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction and filtering to obtain a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:25; the amount of neutral cellulase added is 1% of the mass of seaweed powder; the amount of pectinase added is 0.4% of the mass of seaweed powder; the amount of papain added is 0.8% of the mass of seaweed powder; the enzymolysis temperature is 53° C., the time is 1.5 h; the high temperature and high pressure extraction pressure is 250 MPa, the temperature is 85° C., and the time is 30 min;

[0059] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0060] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0061] Comparative Example 1

[0062] This comparative example is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0063] (a) washing and drying the seaweed until the water content of the seaweed is 9.4%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0064] (b) placing seaweed powder, neutral cellulase and pectinase in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction and filtering to obtain a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:25; the amount of neutral cellulase added is 1% of the mass of the seaweed powder; the amount of pectinase added is 0.4% of the mass of the seaweed powder; the enzymolysis temperature is 53° C., the time is 1.5 h; the high temperature and high pressure extraction pressure is 250 MPa, the temperature is 85° C., and the time is 30 min;

[0065] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0066] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0067] Comparative Example 2

[0068] This comparative example is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0069] (a) washing and drying the seaweed until the water content of the seaweed is 9.4%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0070] (b) placing seaweed powder, pectinase and papain in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction and filtering to obtain a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:25; the amount of pectinase added is 0.4% of the mass of the seaweed powder; the amount of papain added is 0.8% of the mass of the seaweed powder; the enzymolysis temperature is 53° C., the time is 1.5 h; the high temperature and high pressure extraction pressure is 250 MPa, the temperature is 85° C., and the time is 30 min;

[0071] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0072] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0073] Comparative Example 3

[0074] This comparative example is a method for extracting seaweed polysaccharides, and the method for extracting seaweed polysaccharides comprises the following steps:

[0075] (a) washing and drying the seaweed until the water content of the seaweed is 9.4%, crushing the seaweed through an 80-mesh sieve, and collecting the sieve residue to obtain seaweed powder;

[0076] (b) placing seaweed powder, neutral cellulase, pectinase and bromelain in water for enzymolysis and enzyme inactivation, then performing high temperature and high pressure extraction, filtering, and obtaining a crude seaweed polysaccharide extract, wherein the mass ratio of seaweed powder to water is 1:25; the amount of neutral cellulase added is 1% of the mass of the seaweed powder; the amount of pectinase added is 0.4% of the mass of the seaweed powder; the amount of bromelain added is 0.8% of the mass of the seaweed powder; the enzymolysis temperature is 53° C., the time is 1.5 h; the high temperature and high pressure extraction pressure is 250 MPa, the temperature is 5085° C., and the time is 30 min;

[0077] (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation (80% ethanol concentration), centrifugation, and anhydrous ethanol washing to obtain a crude seaweed polysaccharide extract;

[0078] (d) The crude seaweed polysaccharide extract is dissolved in water, and then decolorized by activated carbon, deproteinized by Sevage method, and desalted by ion dialysis method, and then precipitated with alcohol (80% ethanol concentration), and dried in vacuum to obtain the seaweed polysaccharide.

[0079] Experimental example

[0080] The extraction experiments of seaweed polysaccharides were carried out according to the extraction methods of Example 5 and Comparative Examples 1 to 3, and the extraction rate of seaweed polysaccharides was calculated and the purity was tested. The calculation results are shown in Table 1:

[0081] Table 1

[0082] Group Extraction rate purity Example 5 96.7％ 99.8％ Comparative Example 1 89.8％ 99.9％ Comparative Example 2 86.5％ 99.2％ Comparative Example 3 97.1％ 87.2％

[0083] From Table 1, we can see that:

[0084] The technical solution of the present application can better extract seaweed polysaccharides from seaweed and improve the purity of the product by limiting the specific process.

[0085] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein by equivalents. These modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention, and they should all be included in the scope of the claims and specification of the present invention.

Claims

1. A method for extracting seaweed polysaccharides, characterized in that: The steps include: (a) washing, drying and crushing seaweed to obtain seaweed powder; (b) placing seaweed powder and neutral cellulase, pectinase and papain in water for enzymolysis, then extracting under high temperature and high pressure, filtering, and obtaining a crude seaweed polysaccharide extract; (c) subjecting the crude seaweed polysaccharide extract to alcohol precipitation, centrifugation, and washing with anhydrous ethanol to obtain a crude seaweed polysaccharide extract; (d) dissolving the crude seaweed polysaccharide extract in water, and then sequentially performing decolorization, deproteinization and desalination treatments, and then performing alcohol precipitation and drying to obtain the seaweed polysaccharide.

2. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The drying step is performed until the water content of the seaweed is 8% to 10%.

3. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The pulverizing step is performed to a particle size of not less than 80 meshes.

4. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The mass ratio of the seaweed powder to water is 1:(20-30).

5. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The amount of the neutral cellulase added is 0.8% to 1.2% of the mass of the seaweed powder; The added amount of the pectinase is 0.2% to 0.5% of the mass of the seaweed powder.

6. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The enzyme activity of the neutral cellulase is not less than 100,000 U / g; The enzymatic activity of the pectinase is not less than 20,000 U / g.

7. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The added amount of the papain is 0.5% to 1% of the mass of the seaweed powder.

8. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The enzyme activity of the papain is not less than 100,000 U / g.

9. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The enzymolysis temperature is 50-55° C. and the time is 1-2 hours.

10. The method for extracting seaweed polysaccharides according to claim 1, characterized in that: The high temperature and high pressure extraction pressure is 200-300 MPa, the temperature is 80-90° C., and the time is 20-40 min.