Haematococcus pluvialis RXXHp1 and application thereof

Abstract

The invention relates to the technical field of microorganisms, and provides Haematococcus pluvialis RXXHp1 which is preserved in the China Center for Type Culture Collection on May 19, 2023, and the preservation number of the Haematococcus pluvialis RXXHp1 is CCTCC NO: M 2023791. The invention also provides an application of the haematococcus pluvialis RXXHp1. The application comprises the step of enriching at least one of organic selenium and astaxanthin in a selenium ion stress environment by using the haematococcus pluvialis RXXHp1. The invention further provides algae powder containing the haematococcus pluvialis RXXHp1 and application of the haematococcus pluvialis RXXHp1 or the algae powder in preparation of wine. The haematococcus pluvialis RXXHp1 provided by the invention has certain tolerance to a selenium ion stress environment, not only can be used for enriching organic selenium, but also can be used for promoting the accumulation of astaxanthin; besides, when the haematococcus pluvialis RXXHp1 is used for preparing the wine, the content of active substances such as astaxanthin and xanthophyll in the wine can be remarkably increased, so that the effects of improving the color and flavor of the wine and improving the oxidation resistance of the wine are achieved.

Description

Technical Field

[0001] The present disclosure relates to the field of microbial technology, for example, to Haematococcus pluvialis RXXHp1 and applications thereof. Background Art

[0002] In recent years, with a significant increase in health awareness, public dietary preferences have gradually shifted, first from "eating enough" to "eating well," and then from "eating well" to "dietary therapy." Against this backdrop, the phenomenon of hidden hunger is particularly concerning. This refers to a subtle but potentially serious form of malnutrition, characterized by a chronic lack of essential vitamins and minerals in the diet despite adequate caloric intake. According to the 2022 Global Nutrition Report, over 2 billion people worldwide suffer from hidden hunger. Among these, selenium, a trace element, is particularly difficult to supplement, making it a critical nutritional shortcoming that needs to be addressed.

[0003] Selenium, an essential nutrient for the human body, plays a crucial role in maintaining immune function, protecting cardiovascular health, and combating cancer. However, selenium is not essential for the growth and development of higher plants, making it difficult to provide a stable, accurate, and high-quality supply through conventional diets of grains, vegetables, and fruits. However, inorganic selenium is not an ideal choice due to its high toxicity. In contrast, organic selenium has gained attention due to its high bioavailability and low toxicity. Organic selenium primarily includes selenoamino acids, selenoproteins, selenopolysaccharides, selenonucleic acids, and various methylated selenium species; selenoproteins are the crown jewel of organic selenium. Research has shown that selenoproteins are primarily found in algae, humans, and animals, and have not yet been found in fungi, higher plants, or yeast. Compared to other species, algae have a higher number of selenoproteins and a richer variety of organic selenium. Microalgae, as the primary absorber of selenium in aquatic ecosystems, can rapidly absorb inorganic selenium from water and convert it into organic selenium, which is more efficiently absorbed by the human body. These microalgae are excellent carriers of selenium fortification. Therefore, to address the problem of selenium deficiency in hidden hunger, using microalgae as the base organism and increasing the concentration of selenoproteins through agronomic biofortification is considered to be a simple, fast and effective solution to address hidden hunger in the human body.

[0004] Haematococcus pluvialis is a single-celled eukaryotic green algae and a new resource food approved by the National Health Commission. As one of the earliest photosynthetic life forms on Earth, Haematococcus pluvialis has survived five mass extinction events. It not only does not compete with humans for food, land, fertilizer, or time, and is not restricted by seasons, but also has the advantages of being industrially scalable, having high energy efficiency, strong stress resistance, high yield, edible whole plants, and high nutritional value. More importantly, Haematococcus pluvialis can synthesize and accumulate astaxanthin, making it a high-quality source of natural astaxanthin. Astaxanthin is a natural tetraterpenoid secondary metabolite with potent antioxidant properties. Its antioxidant capacity is 500 times that of vitamin E and 6,000 times that of vitamin C. It also has the ability to scavenge free radicals efficiently, and is widely used in food, medicine, feed, cosmetics, and other fields.

[0005] For example, Chinese patent document CN202210992437.2 provides a method for heterotrophically cultivating microalgae products with high organic selenium content. This method improves selenium tolerance and enrichment capacity by targeted domestication of heterotrophic Chlorella. It has the advantages of simplicity, convenience, low cost, and a wide range of applications. The domesticated algae strains can be heterotrophically cultivated to obtain Chlorella products with high organic selenium content and high density. It can be seen that the above patent document only discloses a method for enriching selenium by domesticating heterotrophic Chlorella, but has not yet addressed Haematococcus pluvialis, which can enrich selenium and even synergistically enrich organic selenium and astaxanthin.

[0006] In summary, there is an urgent need for a Haematococcus pluvialis that can enrich organic selenium, or even synergistically enrich organic selenium and astaxanthin. Summary of the Invention

[0007] The purpose of the present disclosure is to overcome the deficiencies in the prior art and provide a Haematococcus pluvialis RXXHp1 and its application. The Haematococcus pluvialis RXXHp1 has a certain tolerance to selenium ion stress environments, can not only enrich organic selenium, but also promote the accumulation of astaxanthin; in addition, the Haematococcus pluvialis RXXHp1 can be used in the preparation of wine to significantly increase the content of active substances such as astaxanthin and lutein in the wine, thereby achieving the effect of improving the color and flavor of the wine and enhancing the antioxidant capacity of the wine.

[0008] The purpose of this disclosure is achieved through the following technical solutions:

[0009] On the one hand, a Haematococcus pluvialis RXXHp1 is provided. The Haematococcus pluvialis RXXHp1 was deposited in the China Center for Type Culture Collection on May 19, 2023, with a preservation number of CCTCC NO: M2023791.

[0010] In another aspect, a use of the Haematococcus pluvialis RXXHp1 as described in the above embodiment is provided, wherein the use comprises: using the Haematococcus pluvialis RXXHp1 to enrich at least one of organic selenium and astaxanthin under selenium ion stress.

[0011] In some embodiments, the application includes: using the Haematococcus pluvialis RXXHp1 to enrich organic selenium and astaxanthin under a selenium ion stress environment.

[0012] In some embodiments, the use includes: when the Haematococcus pluvialis RXXHp1 is used to enrich organic selenium and astaxanthin under a selenium ion stress environment, the Haematococcus pluvialis RXXHp1 is subjected to mixotrophic cultivation.

[0013] In some examples, the conditions for the mixotrophic culture include: a culture temperature of 24-26°C, a culture speed of 150-180 rpm, and a light intensity of 100-500 μmol·m -2 ·s -1 The culture time is 5 to 9 days.

[0014] Exemplarily, the conditions for the mixotrophic culture include: a culture temperature of 24-26°C, a culture speed of 180 rpm, and a light intensity of 300 μmol·m -2 ·s -1 , the culture time is 7 days.

[0015] In some embodiments, in the selenium ion stress environment, the concentration of selenium ions is less than or equal to 1500 mg / L.

[0016] In some examples, in the selenium ion stress environment, the concentration of selenium ions is 100-1500 mg / L.

[0017] Exemplarily, in the selenium ion stress environment, the concentration of selenium ions is 1500 mg / L.

[0018] It is worth noting that the Haematococcus pluvialis RXXHp1 provided by the present disclosure has a certain tolerance to selenium ion stress environment, so that it can not only enrich organic selenium in a selenium ion stress environment, but also promote the accumulation of astaxanthin, and has the effect of synergistically enriching organic selenium and astaxanthin.

[0019] In another aspect, a use of the Haematococcus pluvialis RXXHp1 as described in the above embodiment is provided, wherein the use comprises: using the Haematococcus pluvialis RXXHp1 to prepare a product rich in organic selenium and / or astaxanthin.

[0020] In some examples, the product includes at least one of a food, a feed, and a cosmetic.

[0021] It should be understood that while Chinese patent document CN202210992437.2 provides a method for enriching selenium using microalgae, the algae species used therein are different from those used in the present disclosure. Specifically, the algae species used in Chinese patent document CN202210992437.2 is Chlorella vulgaris, while the algae species used in the present disclosure is Haematococcus pluvialis. Therefore, Chinese patent document CN202210992437.2 does not constitute technical inspiration for the present disclosure.

[0022] In another aspect, an algal powder is provided, comprising: Haematococcus pluvialis RXXHp1 as described in the above embodiment.

[0023] In another aspect, a method for preparing the algal powder described in the above embodiment is provided. The method comprises: inoculating Haematococcus pluvialis RXXHp1 into a modified BG11 liquid culture medium supplemented with ampicillin and cephalosporin for expansion to obtain an algal liquid; settling and separating the algal liquid to obtain an algal mud; and drying the algal mud to obtain the algal powder.

[0024] In some examples, the formula of the modified BG11 liquid culture medium is: sodium acetate 1.0-3.0 g / L, sodium nitrate 1.4-1.6 g / L, K2HPO4·3H2O 0.03-0.05 g / L, MgSO4·7H2O 0.065-0.085 g / L, CaCl2·2H2O 0.026-0.046 g / L, citric acid 0.005-0.007 g / L, ammonium ferric citrate 0.005-0.007 g / L, EDTA 0.0005-0.0015 g / L, sodium carbonate 0.015-0.025 g / L, boric acid 0.0027-0.0029 g / L, MnCl2·H2O 0.0017~0.0019g / L, ZnSO4·7H2O0.00021~0.00023g / L, CuSO4·5H2O 0.00007~0.00009g / L, Na2MoO4·2H2O 0.0003~0.0005g / L and Co(NO3)2·6H2O 0.00004~0.00006g / L; the pH value of the improved BG11 liquid culture medium is 6.8~7.2.

[0025] Exemplarily, the concentration of ampicillin is 10-40 mg / L.

[0026] Exemplarily, the concentration of cephalosporin is 25-75 mg / L.

[0027] In some examples, the conditions for the expanded culture include: a light-dark cycle of 8 to 16 h / 8 to 16 h, a light intensity of 30 to 70 μmol / m -2 / s -1 , the culture temperature is 20-28°C, and the culture speed is 150-200rpm.

[0028] In some examples, the settling includes natural settling.

[0029] Exemplarily, the natural sedimentation time is 2 to 3 days.

[0030] In some examples, the drying comprises spray drying.

[0031] Exemplarily, the spray drying temperature is 150-200°C.

[0032] On the other hand, the present invention provides a use of the Haematococcus pluvialis RXXHp1 as described in the above embodiment in the preparation of wine.

[0033] In some examples, the wine includes at least one of an infused wine and an liqueur wine.

[0034] In another aspect, a use of the algae powder described in the above embodiment or the method described in the above embodiment in the preparation of wine is provided.

[0035] In some embodiments, the wine comprises at least one of an infused wine and an liqueur wine.

[0036] In some examples, the method for preparing the soaking wine includes: mixing the algae powder with a white wine base wine, performing extraction, and obtaining the soaking wine.

[0037] Exemplarily, the mass of the algae powder accounts for 0.1% to 5% of the mass of the liquor base.

[0038] Exemplarily, the liquor base includes light-fragrance kaoliang liquor.

[0039] For example, the alcohol content of the light-fragrance kaoliang liquor is 42% vol to 100% vol.

[0040] Exemplarily, the extraction is carried out at room temperature in the dark.

[0041] For example, the extraction time is 7 to 100 days.

[0042] In some examples, the method for preparing the liquor includes: mixing the algae powder with a white wine base, homogenizing, soaking, and filtering to obtain the liquor.

[0043] Exemplarily, the mass of the algae powder accounts for 0.1% to 5% of the mass of the liquor base.

[0044] Exemplarily, the liquor base includes light-fragrance kaoliang liquor.

[0045] For example, the alcohol content of the light-fragrance kaoliang liquor is 42% vol to 100% vol.

[0046] Exemplarily, the homogenizing comprises high-pressure homogenizing.

[0047] For example, the pressure of the high-pressure homogenization is 30-40 MPa.

[0048] Exemplarily, the immersion is performed under light-proof conditions.

[0049] For example, the soaking temperature is 15-20° C., and the soaking time is 7-100 days.

[0050] The beneficial effects of the present disclosure are:

[0051] 1. The present invention provides a Haematococcus pluvialis RXXHp1 that has a certain tolerance to selenium ion stress environment, so that it can not only enrich organic selenium in a selenium ion stress environment, but also promote the accumulation of astaxanthin, and has the effect of synergistically enriching organic selenium and astaxanthin.

[0052] 2. The Haematococcus pluvialis RXXHp1 provided in the present disclosure can not only be used for the preparation of wine, but also can significantly increase the content of active substances such as astaxanthin and lutein in the wine, thereby achieving the effect of improving the color and flavor of the wine and enhancing the antioxidant capacity of the wine.

[0053] Biological Deposits

[0054] The present invention provides a Haematococcus pluvialis RXXHp1, which was deposited in the China Center for Type Culture Collection (CCTCC) on May 19, 2023, with a deposit number of CCTCC NO: M 2023791. The deposit address is Wuhan University, No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, with a postal code of 430072. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 This is a schematic diagram of the morphological changes of algal cells of Haematococcus pluvialis in Part 5 of Example 2;

[0056] Figure 2 This is a BLAST sequence alignment result of the 18S rRNA gene sequence of Haematococcus pluvialis in Part 5 of Example 2 in the NCBI database;

[0057] Figure 3This is a BLAST sequence alignment result of the RbcL gene sequence of Haematococcus pluvialis in Part 5 of Example 2 in the NCBI database;

[0058] Figure 4 This is a BLAST sequence alignment result of the tufA gene sequence of Haematococcus pluvialis in Part 5 of Example 2 in the NCBI database;

[0059] Figure 5 The neighbor-joining tree constructed based on the 18S rRNA gene sequence of Haematococcus pluvialis in Part 5 of Example 2;

[0060] Figure 6 The neighbor-joining tree constructed based on the RbcL gene sequence of Haematococcus pluvialis in Part 5 of Example 2;

[0061] Figure 7 The neighbor-joining tree constructed based on the tufA gene sequence of Haematococcus pluvialis in Part 5 of Example 2;

[0062] Figure 8 This is a graph showing the effects of different concentrations of sodium selenite treatment on the macroscopic phenotype of RXXHp1 heterotrophically cultured cells in Part 2 of Example 3;

[0063] Figure 9 This is a graph showing the effects of different concentrations of sodium selenite treatment on RXXHp1 heterotrophically cultured microscopic cells in Part 2 of Example 3;

[0064] Figure 10 This is a graph showing the effect of different concentrations of sodium selenite treatment on the optical density of RXXHp1 heterotrophically cultured cells in Part 2 of Example 3;

[0065] Figure 11 This is a graph showing the effect of different concentrations of sodium selenite treatment on chlorophyll fluorescence in heterotrophic culture of RXXHp1 in Part 2 of Example 3;

[0066] Figure 12 This is a graph showing the effect of different concentrations of sodium selenite treatment on the maximum photosynthetic efficiency and actual photosynthetic efficiency of heterotrophic culture of RXXHp1 in Part 2 of Example 3; wherein, Figure 12 A shows the changes in the maximum photosynthetic efficiency of RXXHp1; Figure 12 B shows the changes in the actual photosynthetic efficiency of RXXHp1;

[0067] Figure 13 This is a graph showing the effect of high concentration sodium selenite treatment on the macroscopic phenotype and chlorophyll fluorescence of heterotrophically cultured RXXHp1 cells in Part 2 of Example 3;

[0068] Figure 14This is a graph showing the effect of high concentration sodium selenite treatment on RXXHp1 heterotrophically cultured microscopic cells in Part 2 of Example 3;

[0069] Figure 15 This is a graph showing the effect of high concentration sodium selenite treatment on the maximum photosynthetic efficiency and actual photosynthetic efficiency of heterotrophic culture of RXXHp1 in Part 2 of Example 3; wherein, Figure 15 A shows the changes in the maximum photosynthetic efficiency of RXXHp1; Figure 15 B shows the changes in the actual photosynthetic efficiency of RXXHp1;

[0070] Figure 16 This is a graph showing the effects of different concentrations of sodium selenite treatment on the macroscopic phenotype of RXXHp1 mixotrophic cultured cells in Part 2 of Example 4;

[0071] Figure 17 This is a graph showing the effect of different concentrations of sodium selenite treatment on the optical density of RXXHp1 mixotrophic culture cells in Part 2 of Example 4;

[0072] Figure 18 This is a graph showing the effect of different concentrations of sodium selenite treatment on chlorophyll fluorescence in mixotrophic culture of RXXHp1 in Part 2 of Example 4;

[0073] Figure 19 This is a graph showing the effect of different concentrations of sodium selenite treatment on the maximum photosynthetic efficiency and actual photosynthetic efficiency of RXXHp1 mixotrophic culture in Part 2 of Example 4; wherein, Figure 19 A shows the changes in the maximum photosynthetic efficiency of RXXHp1; Figure 19 B shows the changes in the actual photosynthetic efficiency of RXXHp1;

[0074] Figure 20 This is a graph showing the effect of different concentrations of sodium selenite treatment on the astaxanthin content of RXXHp1 mixotrophic culture in Part 2 of Example 4;

[0075] Figure 21 This is a graph showing the effect of different concentrations of sodium selenite treatment on the organic selenium content in the mixotrophic culture of RXXHp1 in Part 2 of Example 4.

[0076] Figure 22 This is a graph showing the effects of different addition amounts of Haematococcus pluvialis RXXHp1 algae powder on the physicochemical properties and active ingredients of 42% vol liquor in Example 5;

[0077] Figure 23 This is a graph showing the effects of different addition amounts of Haematococcus pluvialis RXXHp1 algae powder on the physicochemical properties and active ingredients of 47% vol liquor in Example 6;

[0078] Figure 24 This is a graph showing the effects of different addition amounts of Haematococcus pluvialis RXXHp1 algae powder on the physicochemical properties and active ingredients of 53% vol liquor in Example 7;

[0079] Figure 25 This is a graph showing the effects of different addition amounts of Haematococcus pluvialis RXXHp1 algae powder on the physicochemical properties and active ingredients of liquor in Example 8;

[0080] Figure 26 This is a graph showing the effects of different addition amounts of Haematococcus pluvialis RXXHp1 algae powder on the physicochemical properties and active ingredients of liquor in Example 8;

[0081] Figure 27 This is a flavor quality analysis chart of the liquor prepared with different addition amounts of Haematococcus pluvialis RXXHp1 algae powder in Example 8;

[0082] Figure 28 This is a graph showing the effects of different addition amounts of Haematococcus pluvialis RXXHp1 algae powder on the physicochemical properties and active ingredients of 75% vol liquor in Example 9;

[0083] Figure 29 This is a diagram showing the effects of different addition amounts of Haematococcus pluvialis RXXHp1 algae powder on the physicochemical properties and active ingredients of 100% vol liquor in Example 10. DETAILED DESCRIPTION

[0084] The following is a clear and complete description of the technical solutions in some embodiments of the present disclosure. Obviously, the embodiments described are only some of the embodiments of the present disclosure, not all of them. Based on the embodiments provided by the present disclosure, all other embodiments obtained by ordinary technicians in this field are within the scope of protection of the present disclosure.

[0085] Unless the context requires otherwise, throughout the specification and claims, the term "comprising" is to be construed in an open, inclusive sense, that is, meaning "including, but not limited to."

[0086] When describing some embodiments, the expression "A and / or B" may be used. It is easy to understand that "A and / or B" includes the following three combinations: only A, only B, and a combination of A and B.

[0087] When describing some embodiments, the expressions "at least one of A, B and C" and "at least one of A, B or C" may be used, both of which have the same meaning and include the following combinations of A, B and C: only A, only B, only C, the combination of A and B, the combination of A and C, the combination of B and C, and the combination of A, B and C.

[0088] Example 1 Culture medium

[0089] 1. BG11 liquid culture medium, its formula is: sodium nitrate 1.4-1.6 g / L, K2HPO4·3H2O 0.03-0.05 g / L, MgSO4·7H2O 0.065-0.085 g / L, CaCl2·2H2O 0.026-0.046 g / L, citric acid 0.005-0.007 g / L, ammonium ferric citrate 0.005-0.007 g / L, EDTA 0.0005-0.0015 g / L, sodium carbonate 0.015-0.025 g / L, boric acid 0.0027-0.0029 g / L, MnCl2·H2O 0.0017-0.0019 g / L, ZnSO4·7H2O 0.00021~0.00023g / L, CuSO4·5H2O0.00007~0.00009g / L, Na2MoO4·2H2O 0.0003~0.0005g / L and Co(NO3)2·6H2O 0.00004~0.00006g / L; pH value is 6.8~7.2.

[0090] 2. BG11 solid plate, its formula is: agar powder 12-17 g / L, sodium nitrate 1.4-1.6 g / L, K2HPO4·3H2O 0.03-0.05 g / L, MgSO4·7H2O 0.065-0.085 g / L, CaCl2·2H2O 0.026-0.046 g / L, citric acid 0.005-0.007 g / L, ammonium ferric citrate 0.005-0.007 g / L, EDTA 0.0005-0.0015 g / L, sodium carbonate 0.015-0.025 g / L, boric acid 0.0027-0.0029 g / L, MnCl2·H2O 0.0017~0.0019g / L, ZnSO4·7H2O0.00021~0.00023g / L, CuSO4·5H2O 0.00007~0.00009g / L, Na2MoO4·2H2O 0.0003~0.0005g / L and Co(NO3)2·6H2O0.00004~0.00006g / L; pH value is 6.8~7.2.

[0091] 3. Improved BG11 liquid culture medium, whose formula is: sodium acetate 1.0-3.0 g / L, sodium nitrate 1.4-1.6 g / L, K2HPO4·3H2O 0.03-0.05 g / L, MgSO4·7H2O 0.065-0.085 g / L, CaCl2·2H2O 0.026-0.046 g / L, citric acid 0.005-0.007 g / L, ammonium ferric citrate 0.005-0.007 g / L, EDTA 0.0005-0.0015 g / L, sodium carbonate 0.015-0.025 g / L, boric acid 0.0027-0.0029 g / L, MnCl2·H2O 0.0017~0.0019g / L, ZnSO4·7H2O0.00021~0.00023g / L, CuSO4·5H2O 0.00007~0.00009g / L, Na2MoO4·2H2O0.0003~0.0005g / L and Co(NO3)2·6H2O 0.00004~0.00006g / L; pH value is 6.8~7.2.

[0092] Example 2 Collection, activation, separation, purification and identification of algae species

[0093] 1. Collection of algae species

[0094] On July 20, 2022, water samples were collected from Tianfu New District, Chengdu City, Sichuan Province (30°24′10″N, 104°7′18″E) using a phytoplankton net with a pore size of 64 μm (i.e., 200 mesh).

[0095] 2. Activation of algae species

[0096] Transfer the collected water sample to a 15 mL centrifuge tube and let it stand for 1 to 2 hours. Gently remove most of the supernatant, retaining 2 to 3 mL of liquid at the bottom. After thorough mixing, shake and incubate at a temperature of 25 ± 1 ° C and a speed of 180 rpm for 6 hours to fully activate the sample.

[0097] 3. Isolation of Algae Species

[0098] 10 μL of the activated sample was dropped onto a glass slide. After confirming the presence of suspected target algae cells under a microscope, a single cell was picked up under a microscope using the capillary siphon separation method. The aspiration, microscopic examination, and dilution were repeated until only a single target algae cell was contained in the water droplet. The sample was then transferred to a 96-well plate containing 100 μL of BG11 liquid culture medium (without antibiotics) and cultured statically. The light-dark cycle was 12 h / 12 ​​h, and the light intensity was 20-30 μmol / m -2 / s -1 The culture temperature was 25±1°C, and BG11 liquid culture medium was appropriately supplemented to about 100 μL during static culture.

[0099] 4. Purification of Algae

[0100] After culturing a single target algae cell for 20 to 30 days, examine the growth of the algae under a microscope. If the algae cell grows well, dilute it to 10 0 , 10 -1 , 10 -2 and 10 -3 The concentration was fully mixed, and 200 μL was taken from each gradient dilution and spread on the plate containing 50 mg / L ampicillin (Amp 50+ )、50mg / L kanamycin (Kanamycin 50+ ) and 100mg / L cephalosporin (Cef 100+ ) on a BG11 solid plate, and the plate was placed in an inverted culture in a constant temperature light incubator with a light-dark cycle of 12 h / 12 ​​h and a light intensity of 10-30 μmol / m -2 / s -1 The culture temperature is 25±1℃, and the plate is inverted and cultured for 7-15 days until a single algae colony grows. The growth of single algae colonies and bacteria on the plate is observed. If there are obvious bacteria on the plate, pick a single algae colony and add it to 200μL BG11 liquid culture medium (containing Amp 100+ 、Kan 50+ , Cef 100+ ) Mix by pipetting and dilute to 10 -1 and 10 -2 Concentration fully mixed, re-coated on BG11 solid plate (also contains Amp 100+ 、Kan 50+ , Cef 100+ ) and cultured on the plate. The algae were purified by plating the plate multiple times until there were no obvious foreign bacteria on the plate and the single algae colony. Then the single algae colony was picked and placed in the modified BG11 liquid medium (containing Amp 25+ , Cef 50+ ) were cultured in a shaker at a temperature of 25 ± 1 °C, a speed of 180 rpm, a light-dark cycle of 12 h / 12 ​​h, and a light intensity of 50 μmol / m -2 / s -1 , to reserve algal cells for subsequent steps.

[0101] 5. Identification of algae species

[0102] 5.1 Morphological observation of algae species

[0103] A small amount of fresh purified algal cells were taken and their morphology was observed under a Leica DM5000 inverted microscope (Leica, Germany). Images were captured using a Leica DFC320 CCD, and differential interference contrast (DIC) and phase contrast (PH) photography were performed.

[0104] The morphological changes of algal cells Figure 1 As shown in FIG, the morphology is described as follows: the plant body is a single cell, the motile cells are green, oval, spherical or pear-shaped, with two flagella of equal length, and the cell wall is clearly separated from the protoplast to form a periplasmic space; as the cell volume increases, the motile cells begin to turn green-brown (red in the middle, green on the outside), and the shape is mainly spherical; as the culture time increases, most cells lose their flagella and produce new walls outside the protoplast to become spherical immobile cells; the immobile cell period can last for several months, during which time the cells can still slowly proliferate and gradually increase in size; the diameter of the immobile cells is 20 to 50 μm, and as the volume increases, most cells no longer retain the original cell wall and periplasmic space except for the thickened new wall, and the cell contents also change from green-brown to bright red or dark red.

[0105] 5.2 Molecular identification of algal species

[0106] The purified algae were centrifuged at 5000 rpm for 5 min to collect the algae, resuspended with sterile water several times to remove the liquid culture medium, and the algae were collected for further molecular identification. The algae genome was extracted using the Ezup column-type plant tissue genomic DNA extraction kit from Sangon Biotech Co., Ltd. according to the instructions; the algae genome was extracted using the New England Biolabs PCR amplification was performed using High-Fidelity DNA Polymerase.

[0107] The 50 μL PCR reaction system is: 25 ng of genomic DNA of RXXHp1 algae, Reaction Buffer 10μL, 10mM dNTP 1μL, 10μM upstream and downstream primers 2.5μL each, Add 0.5 μL of High-Fidelity DNA Polymerase and make up to 50 μL with double-distilled water.

[0108] PCR molecular identification was performed using three genes: 18S ribosomal RNA gene (18S rRNA), ribulose-1,5-bisphosphate carboxylase / oxygenase large subunit (RbcL), and translation elongation factor Tu (tufA). The reaction conditions were: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 1 min, annealing at 55°C for 30 s, extension at 72°C for 30 s, 35 cycles; extension at 72°C for 5 min.

[0109] Among them, there are three pairs of molecular identification primers, namely:

[0110] 1) Upstream primer 5'-AACCTGGTTGATCCTGCCAGT-3' and downstream primer 5'-TGATCCTTCTGCAGGTTCACCTAC-3' for amplifying the 18S rRNA region;

[0111] 2) amplification of the RbcL region using the upstream primer 5′-CAACCAGGTGTTCCASCTGAAG-3′ and the downstream primer 5′-CTAAAGCTGGCATGTGCCATAC-3′;

[0112] 3) The upstream primer 5'-TGAAACAGAAMAWCGTCATTATGC-3' and the downstream primer 5'-CCTTCNCGAATMGCRAAWCGC-3' were used to amplify the tufA region.

[0113] The products of 18S rRNA, RcbL and tufA amplified by PCR were recovered and purified using Takara gel recovery kit and sent to Chengdu Branch of Sangon Biotech (Shanghai) Co., Ltd. for sequencing. The sequencing results were submitted to the National Center for Biotechnology Information (NCBI) database for Nucleotide BLSAT sequence alignment. It was found that 18S rRNA (such as Figure 2 As shown), RbcL (as Figure 3 As shown) and tufA (as Figure 4 The gene sequencing results of (shown in Figure 2) had a query coverage (Query Cover) of 97%, 98%, and 97% with the corresponding sequences of Haematococcus pluvialis, and a consistency (Per.Ident) of 98.38%, 99.69%, and 99.57%, respectively, and the E value (Evalue) was 0.

[0114] The nucleotide sequences of 18S rRNA, RbcL and tufA of the algae strain are shown in SEQ ID No. 1, SEQ ID No. 2 and SEQ ID No. 3, respectively, and the joint neighbor-joining phylogenetic trees of the gene sequences of 18S rRNA, RbcL and tufA of the algae strain are shown in Figures 5-7 shown.

[0115] Analysis showed that the algae strain was in the same evolutionary branch as Haematococcus pluvialis.

[0116] 6. Preservation of algae species

[0117] Based on the results of comprehensive morphological observation and molecular identification, the algae strain obtained in this example is Haematococcus pluvialis, and the algae strain is identified as Haematococcus pluvialis RXXHp1, which was deposited in the China Center for Type Culture Collection (CCTCC) on May 19, 2023. The deposit address is Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province, Postal Code 430072, and its deposit number is CCTCC NO: M 2023791.

[0118] Example 3 Selenium tolerance test of Haematococcus pluvialis RXXHp1

[0119] 1. Experimental Methods

[0120] 1.1 Treatment with different concentrations of sodium selenite (i.e., Na2SeO3)

[0121] The Haematococcus pluvialis RXXHp1 (i.e., RXXHp1) was inoculated into a modified BG11 liquid medium containing 25 mg / L ampicillin and 50 mg / L cephalosporin for step-by-step culture on a shaker. The culture temperature was 25 ± 1 °C, the culture speed was 180 rpm, the light-dark cycle was 12 h / 12 ​​h, and the light intensity was 50 μmol·m -2 ·s -1 When Haematococcus pluvialis RXXHp1 was cultured to the late logarithmic growth phase, fresh modified BG11 liquid culture medium was replaced, and the initial concentration of algal cells was adjusted to OD680 = 0.2, and then divided into 50 mL Erlenmeyer flasks for subsequent treatment experiments.

[0122] The cells were treated with different concentrations of Na2SeO3, namely 0 mg / L, 100 mg / L, 200 mg / L, 500 mg / L, 1000 mg / L, 1500 mg / L, 2000 mg / L and 3000 mg / L. The culture temperature was 25±1℃, the culture speed was 180 rpm, and the heterotrophic culture was carried out without light for 7 consecutive days.

[0123] 1.2 Determination of microalgae growth curve

[0124] A certain amount of fresh algae liquid was taken, and the optical density of RXXHp1 cells at 680 nm was measured using a microplate reader (Metash, UV-9000, China), and a growth curve was drawn.

[0125] 1.3 Chlorophyll fluorescence analysis

[0126] Chlorophyll fluorescence was measured using MAXI-IMAGING-PAM (WALZ, Germany). After mixing the samples, 200 μL was pipetted into a black 96-well plate and the chlorophyll fluorescence parameters were measured after dark adaptation for 5 minutes.

[0127] 2. Experimental Results

[0128] 2.1 RXXHp1 has a certain tolerance to Na2SeO3 stress

[0129] This example studies the effects of different concentrations of Na2SeO3 treatment (0 mg / L, 100 mg / L, 200 mg / L, 500 mg / L, 1000 mg / L, 1500 mg / L, 2000 mg / L and 3000 mg / L) on the growth characteristics of RXXHp1 algae cells under heterotrophic, light-free culture conditions for 7 days. The results are shown in Figure 2. Figures 8 to 15 The specific performance is as follows:

[0130] Compared with the 0 mg / L Na2SeO3 treatment, the color of the RXXHp1 culture medium changed from green to light yellow with the increase of treatment concentration in the 100-1500 mg / L Na2SeO3 treatment (e.g. Figure 8 The color of algal cells changes from green to light brown, and some cells turn red (as shown in Figure 9 As shown in Figure 2 ); the cell optical density gradually increased with the extension of treatment time (as ... Figure 10 ); Chlorophyll fluorescence changes from blue-green to orange (as shown); Figure 11 As shown in Figure 2), the maximum photosynthetic efficiency Fv / Fm and the actual photosynthetic efficiency Y(II) gradually decreased with the extension of treatment time (as shown in Figure 2). Figure 12 A and 12B);

[0131] The significant difference is that after treatment with high concentrations of Na2SeO3 (2000 mg / L and 3000 mg / L respectively), the color of the culture medium of RXXHp1 turned into light yellow (as shown in Figure 2). Figure 13 The color of algal cells changes to reddish brown, the number of red cells is high, and some cells die (as shown in Figure 14 Chlorophyll fluorescence imaging can hardly detect values ​​on the 5th to 7th day of treatment (as shown in Figure 13 The maximum photosynthetic efficiency Fv / Fm and the actual photosynthetic efficiency Y(II) decreased most significantly. On the 5th to 7th day, the actual photosynthetic efficiency Y(II) was almost undetectable (as shown in Figure 2). Figure 15 A and 15B).

[0132] In summary, RXXHp1 has a certain degree of tolerance to Na2SeO3 concentrations less than or equal to 1500 mg / L.

[0133] Example 4 Test of the effect of synergistic enrichment of organic selenium and astaxanthin by Haematococcus pluvialis RXXHp1

[0134] 1. Experimental Methods

[0135] 1.1 Treatment with different concentrations of sodium selenite (i.e., Na2SeO3)

[0136] The Haematococcus pluvialis RXXHp1 (i.e., RXXHp1) was inoculated into a modified BG11 liquid medium containing 25 mg / L ampicillin and 50 mg / L cephalosporin for step-by-step culture on a shaker. The culture temperature was 25 ± 1 °C, the culture speed was 180 rpm, the light-dark cycle was 12 h / 12 ​​h, and the light intensity was 50 μmol·m -2 ·s -1 When Haematococcus pluvialis RXXHp1 was cultured to the late logarithmic growth phase, fresh modified BG11 liquid culture medium was replaced, and the initial concentration of algal cells was adjusted to OD680 = 0.2, and then divided into 50 mL Erlenmeyer flasks for subsequent treatment experiments.

[0137] Different concentrations of Na2SeO3 were treated, namely 0 mg / L, 100 mg / L, 200 mg / L, 500 mg / L, 1000 mg / L and 1500 mg / L, the culture temperature was 25±1℃, the culture speed was 180 rpm, and the concentration of 300 μmol·m -2 ·s -1 The cells were cultured continuously under illumination for 7 days.

[0138] 1.2 Determination of carotenoid content

[0139] Determination of carotenoid content: 50 ± 5 mg of freeze-dried algae powder was weighed and astaxanthin and other carotenoids were extracted according to the national standard (GB / T311520-2015). Carotenoids were detected by HPLC-DAD and LC-QTOF-MS, and quantified using a standard substance purchased from Sigma-Aldrich.

[0140] 1.3 Determination of selenate and selenite ions

[0141] Weigh 100 ± 1 mg of freeze-dried algae powder, and determine and calculate the selenate and selenite contents by high performance liquid chromatography-inductively coupled plasma mass spectrometry according to the national supply and marketing cooperative industry standard (GH / T 1429-2023).

[0142] 1.4 Determination of total selenium content

[0143] Weigh 100 ± 1 mg of freeze-dried algae powder and determine and calculate the total selenium content using ICP-MS, Method 1, according to the national standard (GB 5009.93-2017). Organic selenium content = total selenium content - (selenate content + selenite content).

[0144] It should be noted that other experimental methods involved in this embodiment are the same as those in Example 2 and will not be repeated here.

[0145] 2. Experimental Results

[0146] 2.1 Effects of different concentrations of Na2SeO3 treatment on the growth of RXXHp1 under light

[0147] The initial concentration of RXXHp1 cells was OD680 = 0.2, 300 μmol·m -2 ·s -1 The results were as follows: Figures 16-17 As shown, the specific performance is: under the treatment of 100-1500 mg / L Na2SeO3, the color of the macroscopic phenotype of the microalgae changes to orange-red (such as Figure 16 The cell optical density value increased significantly, and the increasing trend of 1500 mg / L Na2SeO3 was relatively slow (as shown in Figure 2). Figure 17 shown).

[0148] The above results show that 100-1500 mg / L Na2SeO3 treatment promoted the accumulation of colored pigments in RXXHp1 algal cells, and its growth rate gradually slowed down with the increase of treatment concentration.

[0149] 2.2 Analysis of the effect of Na2SeO3 treatment on chlorophyll fluorescence of RXXHp1 under light

[0150] The results of the effects of different concentrations of Na2SeO3 on the chlorophyll fluorescence parameters of RXXHp1 under light are shown in the figure. Figures 18-19 As shown in Figure 3, compared with 0 mg / L Na2SeO3, under 100-1500 mg / L Na2SeO3 treatment, the chlorophyll fluorescence of RXXHp1 changed from blue-green to yellow-brown on the 0th to 5th day, and was alleviated on the 7th day (as shown in Figure 3). Figure 18 The maximum photosynthetic efficiency Fv / Fm and the actual photosynthetic efficiency Y(II) showed a downward trend from day 0 to day 5, but increased from day 5 to day 7 (as shown in Figure 19 A and 19B).

[0151] The above results indicate that RXXHp1 has the function of adaptive self-regulation and recovery after being subjected to selenium stress.

[0152] 2.3 Effects of different concentrations of Na2SeO3 treatment on the enrichment of organic selenium and astaxanthin by RXXHp1 under light

[0153] The results of the effects of different concentrations of Na2SeO3 treatment on the enrichment of organic selenium and astaxanthin by RXXHp1 under light are shown in the figure. Figures 20-21 As shown in the figure, the specific performance is as follows: the test results of astaxanthin content are as follows Figure 20 As shown in Figure 2, under light, the astaxanthin accumulation of RXXHp1 gradually increased with the increase of Na2SeO3 treatment concentration. After 7 days of treatment with 1500 mg / L Na2SeO3, the astaxanthin content reached 10.53 mg / g. The test results of organic selenium content are shown in Figure 2. Figure 21 As shown in the results, under light, the organic selenium content of RXXHp1 gradually increased with the increase of Na2SeO3 treatment concentration. After 7 days of treatment with 1500 mg / L Na2SeO3, the organic selenium content reached 739 mg / kg, and no inorganic selenium content was detected, indicating that RXXHp1 had a good effect in enriching organic selenium.

[0154] It should be noted that the astaxanthin content and organic selenium content in this example are only the data obtained by mixotrophic cultivation of RXXHp1 for 7 days under treatment with different concentrations of Na2SeO3; it should be understood that as the cultivation time increases, the astaxanthin content and organic selenium content will also increase accordingly.

[0155] In summary, Haematococcus pluvialis RXXHp1 can promote the accumulation of astaxanthin while enriching organic selenium.

[0156] Example 5 Application of Haematococcus pluvialis RXXHp1 in the Preparation of 42% vol Liquor

[0157] The algae liquid of Haematococcus pluvialis RXXHp1 after step-by-step expansion culture on a shaker in Example 2 was naturally settled for 2 to 3 days and then separated, the algae mud was collected, and then the algae mud was spray-dried using a spray dryer (temperature of 150 to 200° C.) to obtain algae powder, which was refrigerated and stored in the dark at a low temperature (temperature of 4 to 10° C.) for later use.

[0158] The preparation method of the above-mentioned dew wine includes: adding algae powder of Haematococcus pluvialis RXXHp1 to 42%vol of a light-fragrant sorghum wine base wine, stirring evenly, homogenizing it once with a high-pressure homogenizer (pressure of 30-40MPa), sealing it, soaking it in the dark at 15-20°C for 7-100d, and filtering it to obtain the wine.

[0159] It should be noted that, in this embodiment, a total of 4 groups of liquor were prepared, in which the mass of the algae powder of Haematococcus pluvialis RXXHp1 accounted for 0%, 1%, 2% and 5% of the mass of the 42% vol light-fragrant kaoliang liquor base liquor, respectively.

[0160] The results are as follows Figure 22 As shown, except for the wine prepared without adding algae powder of Haematococcus pluvialis RXXHp1, the wines prepared in the remaining groups all effectively extracted the red substance in Haematococcus pluvialis. The higher the concentration of algae powder added, the more obvious the red color was. The taste of the wine had both fresh fragrance and algae fragrance, and the astaxanthin content and free radical scavenging ability in the wine also increased with the increase of the concentration of algae powder added.

[0161] Therefore, Haematococcus pluvialis RXXHp1 can not only effectively improve the color of 42%vol wine, but also increase its astaxanthin content, thereby enhancing its free radical scavenging ability.

[0162] Example 6 Application of Haematococcus pluvialis RXXHp1 in the Preparation of 47% vol Liquor

[0163] The algae liquid of Haematococcus pluvialis RXXHp1 after step-by-step expansion culture on a shaker in Example 2 was naturally settled for 2 to 3 days and then separated, the algae mud was collected, and then the algae mud was spray-dried using a spray dryer (temperature of 150 to 200° C.) to obtain algae powder, which was refrigerated and stored in the dark at a low temperature (temperature of 4 to 10° C.) for later use.

[0164] The preparation method of the above-mentioned dew wine includes: adding algae powder of Haematococcus pluvialis RXXHp1 to 47%vol of a light-fragrant sorghum wine base wine, stirring evenly, homogenizing it once with a high-pressure homogenizer (pressure of 30-40MPa), sealing it, soaking it in the dark at 15-20°C for 7-100d, and filtering it to obtain the wine.

[0165] It should be noted that, in this embodiment, a total of 4 groups of liquor were prepared, in which the mass of the algae powder of Haematococcus pluvialis RXXHp1 accounted for 0%, 1%, 2% and 5% of the mass of the 47% vol light-fragrant kaoliang liquor base liquor, respectively.

[0166] The results are as follows Figure 23 As shown, except for the wine prepared without adding algae powder of Haematococcus pluvialis RXXHp1, the wines prepared in the remaining groups all effectively extracted the red substance in Haematococcus pluvialis. The higher the concentration of algae powder added, the more obvious the red color was. The taste of the wine had both fresh fragrance and algae fragrance, and the astaxanthin content and free radical scavenging ability in the wine also increased with the increase of the concentration of algae powder added.

[0167] Therefore, Haematococcus pluvialis RXXHp1 can not only effectively improve the color of 47%vol wine, but also increase its astaxanthin content, thereby enhancing its free radical scavenging ability.

[0168] Example 7 Application of Haematococcus pluvialis RXXHp1 in the Preparation of 53% vol Liquor

[0169] The algae liquid of Haematococcus pluvialis RXXHp1 after step-by-step expansion culture on a shaker in Example 2 was naturally settled for 2 to 3 days and then separated, the algae mud was collected, and then the algae mud was spray-dried using a spray dryer (temperature of 150 to 200° C.) to obtain algae powder, which was refrigerated and stored in the dark at a low temperature (temperature of 4 to 10° C.) for later use.

[0170] The preparation method of the above-mentioned dew wine includes: adding algae powder of Haematococcus pluvialis RXXHp1 to 53%vol of fragrant sorghum wine base wine, stirring evenly, homogenizing it once with a high-pressure homogenizer (pressure is 30-40MPa), sealing it, soaking it in the dark at 15-20℃ for 7-100d, and filtering it to obtain the wine.

[0171] It should be noted that, in this embodiment, a total of 4 groups of liquor were prepared, in which the mass of the algae powder of Haematococcus pluvialis RXXHp1 accounted for 0%, 1%, 2% and 5% of the mass of the 53% vol light-fragrant kaoliang liquor base liquor, respectively.

[0172] The results are as follows Figure 24 As shown, except for the wine prepared without adding algae powder of Haematococcus pluvialis RXXHp1, the wines prepared in the remaining groups all effectively extracted the red substance in Haematococcus pluvialis. The higher the concentration of algae powder added, the more obvious the red color was. The taste of the wine had both fresh fragrance and algae fragrance, and the astaxanthin content and free radical scavenging ability in the wine also increased with the increase of the concentration of algae powder added.

[0173] Therefore, Haematococcus pluvialis RXXHp1 can not only effectively improve the color of 53%vol wine, but also increase its astaxanthin content, thereby enhancing its free radical scavenging ability.

[0174] Example 8 Application of Haematococcus pluvialis RXXHp1 in the Preparation of Liquor

[0175] The physicochemical properties, bioactivity, and flavor of the wines prepared from different amounts of Haematococcus pluvialis RXXHp1 algae powder (HPSW) were comprehensively evaluated and analyzed. The results are as follows:

[0176] With the increase of algae powder addition, Figure 25 As shown in (A), the color of the wine gradually becomes darker; Figure 25 As shown in (B), the pH of the wine gradually increases and becomes weakly acidic; Figure 25 As shown in (C), the total acid content of wine gradually increases; Figure 25 As shown in (D), in terms of alcohol content, at a 1% addition amount, the alcohol content of the wine is reduced to 44 degrees; Figure 25 As shown in (E) and (F), the addition of algae powder reduced the content of reducing sugars and total esters; Figure 25 As shown in (G) and (H), in terms of active ingredients, the addition of algae powder can significantly increase the content of two active ingredients, total flavonoids and total polyphenols.

[0177] like Figure 26 As shown in (A), with the increase of the amount of algae powder added, the antioxidant activity indicators of wine such as DPPH, ABTS, and FRAP were significantly improved; Figure 26 As shown in (B), compared to baijiu (channel 5), rooibos exhibits superior DNA-protective activity, mitigating DNA oxidation in the highly oxidative environment of Fenton's reagent. These results demonstrate that rooibos prepared using Haematococcus pluvialis RXXHp1 algae powder exhibits excellent antioxidant and DNA-protective bioactivities, making it healthier for the human body.

[0178] In addition, electronic noses and electronic tongues can simulate human senses and reflect the flavor quality of food. Figure 27 As shown, the addition of algal powder resulted in a difference in aroma between baijiu and 0.1% HPSW, 0.5% HPSW, and 1% HPSW, with the most significant difference. Furthermore, the electronic tongue demonstrated distinct tastes between baijiu, 0.1% HPSW, 0.5% HPSW, and 1% HPSW, with the 1% HPSW liquor exhibiting stronger sweetness and bitterness. These data demonstrate that the addition of algal powder to 1% HPSW improves the flavor quality of baijiu.

[0179] Example 9 Application of Haematococcus pluvialis RXXHp1 in the Preparation of 75% vol Liquor

[0180] The algae liquid of Haematococcus pluvialis RXXHp1 after step-by-step expansion culture on a shaker in Example 2 was naturally settled for 2 to 3 days and then separated, the algae mud was collected, and then the algae mud was spray-dried using a spray dryer (temperature of 150 to 200° C.) to obtain algae powder, which was refrigerated and stored in the dark at a low temperature (temperature of 4 to 10° C.) for later use.

[0181] The preparation method of the above-mentioned dew wine includes: adding algae powder of Haematococcus pluvialis RXXHp1 to 75% vol of a light-fragrant sorghum wine base wine, stirring evenly, homogenizing it once with a high-pressure homogenizer (pressure of 30-40 MPa), sealing it, soaking it in the dark at 15-20°C for 7-100 days, and filtering it to obtain the wine.

[0182] It should be noted that, in this embodiment, a total of 8 groups of liquor were prepared, in which the mass of the algae powder of Haematococcus pluvialis RXXHp1 accounted for 0%, 0.075%, 0.1%, 0.15%, 0.25%, 0.4%, 1% and 5% of the mass of the 75% vol light-fragrant kaoliang liquor base liquor, respectively.

[0183] The results are as follows Figure 28 As shown, except for the wine prepared without adding algae powder of Haematococcus pluvialis RXXHp1, the wines prepared in the remaining groups all effectively extracted the red substance in Haematococcus pluvialis. The higher the concentration of algae powder added, the more obvious the red color was. The taste of the wine had both fresh fragrance and algae fragrance, and the content of astaxanthin and lutein in the wine and the free radical scavenging ability of the wine also increased with the increase of the concentration of algae powder added.

[0184] Therefore, Haematococcus pluvialis RXXHp1 can not only effectively improve the color of 75%vol wine, but also increase its astaxanthin and lutein contents, thereby enhancing its free radical scavenging ability.

[0185] Example 10 Application of Haematococcus pluvialis RXXHp1 in the Preparation of 100% vol Liquor

[0186] The algae liquid of Haematococcus pluvialis RXXHp1 after step-by-step expansion culture on a shaker in Example 2 was naturally settled for 2 to 3 days and then separated, the algae mud was collected, and then the algae mud was spray-dried using a spray dryer (temperature of 150 to 200° C.) to obtain algae powder, which was refrigerated and stored in the dark at a low temperature (temperature of 4 to 10° C.) for later use.

[0187] The preparation method of the above-mentioned dew wine includes: adding algae powder of Haematococcus pluvialis RXXHp1 to 100% vol of a light-fragrant sorghum wine base wine, stirring evenly, homogenizing it once with a high-pressure homogenizer (pressure of 30-40 MPa), sealing it, soaking it in the dark at 15-20°C for 7-100 days, and filtering it to obtain the wine.

[0188] It should be noted that, in this embodiment, a total of 8 groups of liquor were prepared, in which the mass of the algae powder of Haematococcus pluvialis RXXHp1 accounted for 0%, 0.075%, 0.1%, 0.15%, 0.25%, 0.4%, 1% and 5% of the mass of the 100% vol light-fragrant kaoliang liquor base liquor, respectively.

[0189] The results are as follows Figure 29 As shown, except for the wine prepared without adding algae powder of Haematococcus pluvialis RXXHp1, the wines prepared in the remaining groups all effectively extracted the red substance in Haematococcus pluvialis. The higher the concentration of algae powder added, the more obvious the red color was. The taste of the wine had both fresh fragrance and algae fragrance, and the content of astaxanthin and lutein in the wine and the free radical scavenging ability of the wine also increased with the increase of the concentration of algae powder added.

[0190] Therefore, Haematococcus pluvialis RXXHp1 can not only effectively improve the color of 100%vol wine, but also increase its astaxanthin and lutein contents, thereby enhancing its free radical scavenging ability.

[0191] In summary, the physicochemical properties, active ingredients, antioxidant activity, DNA protection activity, and flavor quality of the liquor developed using the algae powder of Haematococcus pluvialis RXXHp1 are different from those of white liquor, and its overall quality has been improved, making it promising as a functional liquor with health value.

[0192] Therefore, the Haematococcus pluvialis RXXHp1 provided in the present disclosure has a certain tolerance to selenium ion stress environment, can not only enrich organic selenium, but also promote the accumulation of astaxanthin; in addition, when the Haematococcus pluvialis RXXHp1 is used in the preparation of wine, the content of active substances such as astaxanthin and lutein in the wine can be significantly increased, thereby achieving the effect of improving the color and flavor of the wine and enhancing the antioxidant capacity of the wine.

[0193] The foregoing description is merely a preferred embodiment of the present disclosure. It should be understood that the present disclosure is not limited to the form disclosed herein and should not be construed as excluding other embodiments. Rather, the present disclosure is applicable to various other combinations, modifications, and environments and can be modified within the scope of the concepts described herein through the above teachings or techniques or knowledge in the relevant fields. Modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the present disclosure are intended to be protected by the claims appended hereto.

Claims

1. A Haematococcus pluvialis RXXHp1, characterized in that The Haematococcus pluvialis RXXHp1 was deposited in the China Center for Type Culture Collection on May 19, 2023, and its preservation number is CCTCC NO: M2023791.

2. A use of the Haematococcus pluvialis RXXHp1 according to claim 1, characterized in that: include: The Haematococcus pluvialis RXXHp1 is used to enrich at least one of organic selenium and astaxanthin under a selenium ion stress environment.

3. The use according to claim 2, characterized in that include: The Haematococcus pluvialis RXXHp1 is used to enrich organic selenium and astaxanthin under a selenium ion stress environment.

4. The use according to claim 3, characterized in that include: When the Haematococcus pluvialis RXXHp1 is used to enrich organic selenium and astaxanthin under a selenium ion stress environment, the Haematococcus pluvialis RXXHp1 is cultured mixotrophically.

5. The use according to claim 3 or 4, characterized in that In the selenium ion stress environment, the concentration of selenium ions is less than or equal to 1500 mg / L.

6. A use of the Haematococcus pluvialis RXXHp1 according to claim 1, characterized in that: include: The Haematococcus pluvialis RXXHp1 is used to prepare a product rich in organic selenium and / or astaxanthin.

7. An algae powder, characterized in that: include: The Haematococcus pluvialis RXXHp1 according to claim 1.

8. A method for preparing the algae powder according to claim 7, characterized in that: include: inoculating the Haematococcus pluvialis RXXHp1 into a modified BG11 liquid culture medium supplemented with ampicillin and cephalosporin, performing expansion culture to obtain an algae liquid; sedimenting and separating the algae liquid to obtain algae mud; as well as The algae mud is dried to obtain the algae powder.

9. Use of the Haematococcus pluvialis RXXHp1 according to claim 1 in the preparation of wine.

10. Use of the algae powder according to claim 7 or the method according to claim 8 in the preparation of wine.

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