Haematococcus pluvialis culture medium and preparation method thereof

Abstract

The invention belongs to the technical field of microalgae culture, and particularly provides a haematococcus pluvialis culture medium and a preparation method thereof.The haematococcus pluvialis culture medium is prepared from 1-3 mg / L of sodium acetate, 0.01-1 mg / L of cobalt nitrate hexahydrate, 0.01-1.6 mg / L of copper sulfate, 0.1-1.2 mg / L of zinc sulfate heptahydrate, 1-3 mg / L of manganese chloride tetrahydrate, 1-10 mg / L of boric acid, 1-30 mg / L of sodium carbonate, 1100-1500 mg / L of sodium nitrate, 0.01-6 mg / L of EDTA, 20-50 mg / L of calcium chloride, 70-100 mg / L of magnesium sulfate heptahydrate, 40-100 mg / L of monopotassium phosphate trihydrate and 0.02 g of inorganic phosphorus; the method comprises the following specific steps: S1, culturing haematococcus pluvialis to a stable phase by a conventional method, inoculating haematococcus pluvialis species into a haematococcus pluvialis culture medium, and culturing; s2, shaking table; s3, culturing for 45 hours; s4, inoculating the seed solution into a haematococcus pluvialis culture medium, and continuously culturing in the shaking table for 6-15 days; the culture medium disclosed by the invention has a good promotion effect on the growth of haematococcus pluvialis and the accumulation of astaxanthin.

Description

Technical Field

[0001] The invention belongs to the technical field of microalgae cultivation, and in particular relates to a Haematococcus pluvialis culture medium and a preparation method thereof. Background Art

[0002] Haematococcus pluvialis is a single-cell green algae mainly distributed in freshwater. It is currently an important source of natural astaxanthin. Its astaxanthin content has great advantages over other sources, such as crustacean extracts, Phaffia rhodozyma, soil Agrobacterium and Chlorella.

[0003] Current research shows that it is rich in astaxanthin, which can reach 5%, much higher than biological sources such as crustaceans and yeast, and is the main potential source of natural astaxanthin extraction. At the same time, compared with chemically synthesized astaxanthin, natural astaxanthin has higher biological activity and safety. Therefore, natural astaxanthin derived from Haematococcus pluvialis has broad application prospects in natural cosmetics and animal husbandry. At the same time, Haematococcus pluvialis is also the latest new resource food approved by my country, which can be directly used as an additive in the processing of health products and functional foods. Due to the high value attributes of Haematococcus pluvialis, the global demand for Haematococcus pluvialis has increased sharply year by year. In 2022, the market size of astaxanthin derived from Haematococcus pluvialis will be US$194.312 billion.

[0004] Compared with traditional culture media, the culture medium provided by the invention has a better promoting effect on the growth of Haematococcus pluvialis and the accumulation of astaxanthin, and the raw materials are easily available, so the invention has a good application prospect. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides a culture medium and a preparation method thereof, which can solve the technical problems of low growth and astaxanthin accumulation of Haematococcus pluvialis caused by traditional culture medium, and improve the astaxanthin production capacity of Haematococcus pluvialis by optimizing the composition ratio of the culture medium.

[0006] The present invention is achieved in that:

[0007] The invention provides a Haematococcus pluvialis culture medium, which comprises 1-3 mg / L of sodium acetate, 0.01-1 mg / L of cobalt nitrate hexahydrate, 0.01-1.6 mg / L of copper sulfate, 0.1-1.2 mg / L of zinc sulfate heptahydrate, 1-3 mg / L of manganese chloride tetrahydrate, 1-10 mg / L of boric acid, 1-30 mg / L of sodium carbonate, 1100-1500 mg / L of sodium nitrate, 0.0.1-6 mg / L of EDTA, 20-50 mg / L of calcium chloride, 70-100 mg / L of magnesium sulfate heptahydrate, 40-100 mg / L of potassium dihydrogen phosphate trihydrate and 0.02 g of inorganic phosphorus.

[0008] The technical effects of a Haematococcus pluvialis culture medium and a preparation method thereof provided by the present invention are as follows: using the Haematococcus pluvialis culture medium for cultivation can increase the astaxanthin production capacity of the Haematococcus pluvialis and have a favorable effect on the active substances in the Haematococcus pluvialis.

[0009] On the basis of the above technical solution, the culture medium of Haematococcus pluvialis and the preparation method thereof of the present invention can also be improved as follows:

[0010] Furthermore, the inorganic phosphorus is KH2PO4.

[0011] Further, the specific steps are as follows: S1: culturing Haematococcus pluvialis to a stable stage by conventional methods, inoculating Haematococcus pluvialis algae seeds with Haematococcus pluvialis culture medium, and culturing;

[0012] S2: Shaking table;

[0013] S3: culture for 45 h;

[0014] S4: The seed solution was inoculated into the culture medium of Haematococcus pluvialis and cultured in a shaking incubator for 6-15 days;

[0015] Furthermore, the pH of the culture medium in S1 is 7-8.

[0016] Furthermore, the shaking table speed in S2 is 200 rpm / min.

[0017] Furthermore, the culture temperature in S3 is 28°C.

[0018] Furthermore, the culture temperature in S4 is 28°C.

[0019] Furthermore, the shaking table speed in S4 is 250 rpm / min.

[0020] Compared with the prior art, the culture medium for Haematococcus pluvialis and the preparation method thereof provided by the present invention have the following beneficial effects: the culture medium has high culture efficiency, and compared with traditional culture medium, it has a better promoting effect on the growth of Haematococcus pluvialis and the accumulation of astaxanthin, and the raw materials are easily available, and it has a good application prospect and can effectively improve the production efficiency and industrial value of Haematococcus pluvialis. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without paying creative work.

[0022] In the accompanying drawings, the components represented by the reference numerals are listed as follows:

[0023] Figure 1 The present invention is a flow chart of a Haematococcus pluvialis culture medium and a preparation method thereof. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0025] Therefore, the following detailed description of the embodiments of the present invention provided in the accompanying drawings is not intended to limit the scope of the invention claimed for protection, but merely represents selected embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] It should be noted that similar reference numerals and letters denote similar items in the following drawings, and therefore, once an item is defined in one drawing, further definition and explanation thereof is not required in subsequent drawings.

[0027] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore should not be understood as a limitation on the present invention.

[0028] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. In the description of the present invention, the meaning of "plurality" is two or more, unless otherwise clearly and specifically defined.

[0029] Example

[0030] As shown in the figure, the present invention provides a culture medium for Haematococcus pluvialis and a preparation method thereof, which includes 1-3 mg / L of sodium acetate, 0.01-1 mg / L of cobalt nitrate hexahydrate, 0.01-1.6 mg / L of copper sulfate, 0.1-1.2 mg / L of zinc sulfate heptahydrate, 1-3 mg / L of manganese chloride tetrahydrate, 1-10 mg / L of boric acid, 1-30 mg / L of sodium carbonate, 1100-1500 mg / L of sodium nitrate, 0.0.1-6 mg / L of EDTA, 20-50 mg / L of calcium chloride, 70-100 of magnesium sulfate heptahydrate, 40-100 of potassium dihydrogen phosphate trihydrate, and 0.02 g of inorganic phosphorus.

[0031] Optionally, in the above technical solution, the inorganic phosphorus is KH2PO4.

[0032] Optionally, in the above technical solution,

[0033] S1: Cultivate the Haematococcus pluvialis to a stable stage using a conventional method, inoculate the Haematococcus pluvialis algae with a Haematococcus pluvialis culture medium for cultivation;

[0034] S2: Shaking table;

[0035] S3: culture for 45 h;

[0036] S4: The seed solution was inoculated into the culture medium of Haematococcus pluvialis and cultured in a shaking incubator for 6-15 days;

[0037] Optionally, in the above technical solution, the pH of the culture medium in S1 is 7-8.

[0038] Optionally, in the above technical solution, the shaking table speed in S2 is 200 rpm / min.

[0039] Optionally, in the above technical solution, the culture temperature in S3 is 28°C

[0040] Optionally, in the above technical solution, the culture temperature in S4 is 28°C.

[0041] Optionally, in the above technical solution, the shaking table speed in S4 is 250 rpm / min.

[0042] Example:

[0043] Haematococcus pluvialis was purchased from the Freshwater Algae Seed Bank of the Institute of Hydrobiology, Chinese Academy of Sciences. The Haematococcus pluvialis species was FACHB-877 and the culture medium was BG11.

[0044] Medium BG611 was set as control medium;

[0045] The components of the control culture medium are: component concentration (mg / L): sodium nitrate 1300; magnesium sulfate heptahydrate 100; potassium hydrogen phosphate trihydrate 40; citric acid monohydrate 10; calcium chloride 30; ferric citrate 5; disodium ethylenediaminetetraacetic acid 5; boric acid 5; manganese sulfate 5; zinc sulfate heptahydrate 0.5; sodium molybdate dihydrate 0.5; copper sulfate pentahydrate 0.05; cobalt chloride 0.05; sodium carbonate 25.

[0046] Among them, the pH of the culture medium BG611 is 7.2.

[0047] The formula of the culture medium of Example 1 is as follows: component concentration (mg / L): 2 mg / L of sodium acetate, 0.02 mg / L of cobalt nitrate hexahydrate, 1 mg / L of copper sulfate, 1.2 mg / L of zinc sulfate heptahydrate, 2 mg / L of manganese chloride tetrahydrate, 10 mg / L of boric acid, 15 mg / L of sodium carbonate, 1100 mg / L of sodium nitrate, 0.5 mg / L of EDTA, 25 mg / L of calcium chloride, 70 mg / L of magnesium sulfate heptahydrate, and 50 mg / L of potassium dihydrogen phosphate trihydrate, wherein the inorganic phosphorus is 0.02 g.

[0048] Wherein, the pH of the culture medium in Example 1 is 7.7.

[0049] After the culture was completed, the astaxanthin content and the cell dry weight of Haematococcus pluvialis were determined;

[0050] The content of astaxanthin was determined by HPLC method; pH was determined by pH detector;

[0051] The control culture medium and the culture medium of Example 1 were set up in triplicate.

[0052] As shown in the table:

[0053] Haematococcus pluvialis dry weight (mg / ml) Astaxanthin content in Haematococcus pluvialis (weight %) Control group 0.11 0.5 Control group 0.12 0.7 Control group 0.11 0.6 Example 1 0.8 1.1 Example 1 1.0 1.3 Example 1 1.1 1.2

[0054] The results show that the dry weight of Haematococcus pluvialis and the astaxanthin content in the Haematococcus pluvialis of the embodiment are higher than those of the control group. Therefore, the Haematococcus pluvialis culture medium of the embodiment can increase the dry weight of Haematococcus pluvialis and the astaxanthin content in the Haematococcus pluvialis, thereby solving the technical problems of the traditional culture medium on the growth of Haematococcus pluvialis and the low accumulation of astaxanthin.

[0055] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A culture medium for Haematococcus pluvialis, characterized in that: Including sodium acetate 1-3mg / L, cobalt nitrate hexahydrate 0.01-1mg / L, copper sulfate 0.01-1.6mg / L, zinc sulfate heptahydrate 0.1-1.2mg / L, manganese chloride tetrahydrate 1-3mg / L, boric acid 1-10mg / L, sodium carbonate 1-30mg / L, sodium nitrate 1100-1500mg / L, EDTA 0.01-6mg / L, calcium chloride 20-50mg / L, magnesium sulfate heptahydrate 70-100mg / , potassium dihydrogen phosphate trihydrate 40-100mg / L, and inorganic phosphorus 0.02g.

2. The method for culturing and preparing Haematococcus pluvialis according to claim 1, characterized in that: The inorganic phosphorus is KH2PO4.

3. A Haematococcus pluvialis culture medium according to claim 1, characterized in that: The specific steps are: S1: Cultivate the Haematococcus pluvialis to a stable stage using a conventional method, inoculate the Haematococcus pluvialis algae with a Haematococcus pluvialis culture medium for cultivation; S2: Shaking table; S3: culture for 45 h; S4: The seed solution is inoculated into the culture medium of Haematococcus pluvialis and cultured in a shaking incubator for 6-15 days.

4. The method for preparing a culture medium of Haematococcus pluvialis according to claim 3, characterized in that: The pH of the culture medium in S1 is 7-8.

5. The method for preparing a culture medium of Haematococcus pluvialis according to claim 3, characterized in that: The shaking speed in S2 is 200 rpm / min.

6. The method for preparing a culture medium of Haematococcus pluvialis according to claim 3, characterized in that: The culture temperature in S3 was 28°C.

7. The method for preparing a culture medium of Haematococcus pluvialis according to claim 3, characterized in that: The culture temperature in S4 is 28°C.

8. The method for preparing a culture medium of Haematococcus pluvialis according to claim 3, characterized in that: The shaking speed in S4 is 250 rpm / min.

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