A cultivation method of gonyostomum semen, an extraction method of active components of gonyostomum semen, and application of the extract in cosmetics

Abstract

The application discloses cultivation of a double-capsule algae, an active component extraction method and application of an extract thereof in cosmetics, and belongs to the field of marine microalgae cosmetics. Geminocystis The double-capsule algae (D. sp.) MH1 is derived from a tidal zone of the South China Sea, and has a preservation number of CCTCC NO: M 2026091. Experiments prove that water extract, alcohol extract and ester extract of the double-capsule algae have significant antioxidant, whitening, anti-inflammatory and sunscreen effects, and are suitable for preparation of cosmetics with antioxidant, whitening, sunscreen and anti-inflammatory four-in-one effects. The algal strain has high cultivation efficiency, the extraction process is simple, active components are rich, a new way is provided for development of natural, four-in-one cosmetic raw materials, and has a wide application prospect.

Description

Technical Field

[0001] This invention relates to the fields of marine microalgae biotechnology and cosmetics, specifically to a species of algae (Dicystis aeruginosa). Geminocystis Methods for the cultivation, harvesting, and extraction of active components of sp. MH1, and the application of its extracts in the preparation of cosmetics and other daily chemical products with antioxidant, whitening, anti-inflammatory, and UV-shielding active ingredients. Background Technology

[0002] Ultraviolet radiation, environmental pollution, and life stress exacerbate skin oxidative stress and photoaging. Traditional chemical sunscreens pose potential risks of skin irritation and environmental cumulative toxicity; while plant-derived active ingredients are generally milder, their activity intensity and stability are often insufficient.

[0003] In recent years, microalgae have become an important source of functional raw materials for cosmetics due to their advantages such as high growth rate, high photosynthetic efficiency, short cultivation cycle, abundant metabolites, small land occupation, and no competition for arable land with traditional agricultural crops. Microalgae are rich in a variety of high-value bioactive substances, including long-chain polyunsaturated fatty acids, polysaccharides, phycobiliproteins, vitamins, carotenoids, and sterols. They possess various activities such as anti-aging, anti-inflammatory, antioxidant, anticancer, antihypertensive, antibacterial, and immunomodulatory effects, and can be used to produce high-value-added chemicals for applications in pharmaceuticals, functional foods, and functional cosmetics.

[0004] Cyanobacteria, as photosynthetic prokaryotes with a long evolutionary history, are widely distributed in various environments, including extreme environments. Their long evolutionary history and survival experience under environmental stress have endowed them with the ability to synthesize protective compounds against external factors such as dryness, ultraviolet radiation, and high salinity. The phylum Cyanobacteria is diverse, and extracts from different species exhibit significant differences in efficacy. This significant difference stems from the different active ingredients contained in different species, leading to unpredictable activity variations in extracts even within the same phylum. For example… Oscillatoria agardhii The extract has some skin-whitening potential. Sacred Aphanotheca The extract has anti-inflammatory properties. Scytonema The extract of sp. has both antioxidant and sun protection activities.

[0005] Besides cyanobacteria, Mirata et al. also studied brown algae in the Mediterranean. Ericaria amentacea Extracts with antioxidant, anti-inflammatory, whitening and photoprotective activities were obtained. Ericaria amentacea Belonging to the phylum Phaeophyceae, it is a large seaweed, a multicellular eukaryote, with a complex thallus structure, typically exceeding 10-15 cm in length, and an apical meristem of 2-3 cm. Although E. amentaceaIt has been proven to possess multiple biological activities, but its biggest current problem is the low biomass yield, making it difficult to meet industrial-scale demands. Due to... E. amentacea As a protected species, its wild resource collection is restricted; moreover, as a large seaweed, its growth and reproduction are strictly controlled by seasonal factors, with doubling time usually measured in days to years, and its biomass accumulation rate is much lower than that of microalgae, and large-scale laboratory cultivation is difficult; in addition, its growth depends on the marine environment, making it difficult to achieve precise process control, resulting in large fluctuations in the content of active ingredients between batches collected at different times and locations, which limits actual production and large-scale application (Mirata et al., Mar. Drugs, 2023, 21,306).

[0006] In contrast, cyanobacteria, as a type of single-celled prokaryotic photosynthetic microorganism, do not rely on complex multicellular tissue differentiation for growth. Instead, they reproduce through simple binary fission, resulting in rapid growth, high biomass yield, no seasonal limitations, continuous year-round production, ease of large-scale laboratory cultivation, strong genetic operability, ease of metabolic engineering modification, precise and controllable culture system, and uniform product quality. These advantages make them a promising candidate to overcome the supply bottleneck of bioactive substances from giant kelp and more feasible for large-scale cultivation and production.

[0007] Dicystis ( Geminocystis *Synechocystis* is a new genus isolated from *Synechocystis* by J. Korelusova et al. in 2009. It belongs to the phylum Cyanophyta, class Cyanophyceae, order Chroococcales, and family Geminocystaceae. In May 2017, Ma Suchao et al. first isolated a strain of *Synechocystis* from Crescent Lake, a closed sub-lake of Qinghai Lake. This was the first reported record of a planktonic *Synechocystis* genus in my country, but research on its active ingredients and functions remains lacking. In recent years, *Synechocystis* has been collected from various regions, including Greece in southeastern Europe, the United States in North America, Brazil in South America, and Japan and South Korea in East Asia. Domestically, apart from Ma Suchao et al.'s first isolation in Qinghai Lake in 2019 and Chen et al.'s rediscovery in Xixi Wetland in Hangzhou in 2025, no other reports have been made of this algae in other regions (especially tropical seas). The habitats and ecological habits of *Dicystis* exhibit a very high degree of diversity, with collection sites including lakes, streams, wetlands, beaches, soil, saline lakes, and mineral water. However, no efficacy reports have been made for extracts from these *Dicystis* species. Therefore, the specific efficacy of *Dicystis* extracts and their potential applications in cosmetics are currently unpredictable. Summary of the Invention

[0008] This invention aims to provide a method for cultivating, harvesting, and extracting active components of *Dicystis jirovecii*, and the application of the extract in the preparation of cosmetics and other daily chemical products. The extract has been experimentally proven to possess multiple biological activities, including antioxidant, whitening, anti-inflammatory, and UV-shielding properties.

[0009] The objective of this invention is achieved through the following technical solution:

[0010] A type of bicysta ( Geminocystis sp.)MH1, deposited at China Center for Type Culture Collection, accession number: CCTCC NO: M 2026091.

[0011] The *Dicystis humicana* MH1 strain originates from the intertidal zone of the South China Sea. As a dominant algal species in tropical nearshore waters, this strain thrives in environments with high temperatures, strong ultraviolet radiation, and drastic salinity fluctuations, and its secondary metabolites are expected to possess multiple therapeutic effects. This invention aims to fill the gap in research on the efficacy of *Dicystis humicana* extracts. Through systematic research on the cultivation, harvesting, and active component extraction methods of *Dicystis humicana*, as well as the various activities of its extracts, this invention reveals the unique efficacy of *Dicystis humicana* extracts compared to other cyanobacteria, providing new directions and options for the development of natural cosmetic raw materials. Compared to traditional cosmetic raw materials, the natural origin of *Dicystis humicana* extract makes it an important subject in the field of cosmetic research. The *Dicystis humicana* extract, originating from the tropical intertidal zone of the South China Sea, possesses four highly effective activities (antioxidant, whitening, sun protection, and anti-inflammatory), achieving a triple effect of "sun protection + repair + whitening." Compared to known cyanobacteria extracts with single or dual activities, this exhibits superior uniqueness and innovation.

[0012] This invention also provides a method for cultivating Dicystis, comprising the following steps:

[0013] 1) Culture Dicystis in a microalgae photobioreactor. The culture medium is F / 2 medium, Zarrouk medium, BG11 medium or modified Dicystis-specific medium SYSU-Lvsan-1. The culture temperature is 20-40℃, the light intensity is 1000-5000 Lux, and the culture time is 1-30 days.

[0014] 2) After the culture is completed, the algal mud is collected by centrifugation after settling for 1-3 days. After centrifugation and washing with deionized water 3 times, the desalinated algal mud is obtained. The algal mud is then freeze-dried to obtain freeze-dried algal powder.

[0015] Furthermore, the photobioreactor is either an intermittent photobioreactor or a continuous flow photobioreactor.

[0016] Furthermore, the formulation of the modified Dicystis-specific culture medium SYSU-Lvsan-1 is as follows: 53.00 mg / L urea, 5.65 mg / L NaH2PO4·H2O, and water as the solvent.

[0017] This invention also provides a method for preparing a Dicystis jirovecii extract, comprising the following steps:

[0018] Add an extraction solvent to algal mud or algal powder, and extract using physical crushing, thermal extraction or organic solvent extraction. Collect the supernatant by centrifugation, and concentrate or freeze-dry to obtain the extract of Dicystis jirovecii.

[0019] The physical fragmentation method includes ultrasonic fragmentation or repeated freeze-thaw cycles;

[0020] The thermal extraction method is boiling extraction;

[0021] The organic solvent extraction method includes alcohol extraction or ester extraction.

[0022] Furthermore, the ultrasonic disruption uses deionized water or buffer solution as the extraction solvent, with an ultrasonic power of 5-650W, an extraction time of 3 min-3 h per extraction, and a total of 1-30 extractions.

[0023] Furthermore, the repeated freeze-thaw cycles use deionized water or buffer solution as the extraction solvent, the freezing temperature is -20°C to -80°C, and the number of freeze-thaw cycles is 1-30.

[0024] Furthermore, the boiling extraction uses deionized water or buffer solution as the extraction solvent, with each extraction lasting 3 minutes to 3 hours, and is repeated 1 to 30 times.

[0025] Furthermore, the alcohol extraction method uses 30%-100% ethanol by volume as the extraction solvent, with each extraction lasting 5 min-72 h, and is repeated 1-30 times.

[0026] Furthermore, the ester extraction method uses ethyl acetate as the extraction solvent, with each extraction lasting 5 min to 72 h, and is repeated 1 to 30 times.

[0027] Specifically, the Dicystis extract of the present invention is prepared by any of the following methods:

[0028] 1) Preparation method of ultrasonic water extract of Dicystis: Add deionized water or buffer to algal mud or algal powder, stir evenly, and extract cells by ultrasonic disruption for 3 min-3 h at a power of 5-650 W. Centrifuge and collect the supernatant. Add an equal amount of deionized water or buffer to the algal body for ultrasonic disruption and extraction. Repeat the above operation 1-30 times, combine the supernatants, and freeze dry to obtain ultrasonic water extract of Dicystis.

[0029] 2) Preparation method of freeze-thaw water extract of Dicystis: Add deionized water or buffer solution to algal mud or algal powder, stir evenly, put it in the freezer, and after freezing is complete, take it out and thaw it, centrifuge and collect the supernatant. Add an equal amount of deionized water or buffer solution to the algal body and extract by ultrasonication. Repeat the above operation 1-30 times, combine the supernatant, and freeze dry to obtain freeze-thaw water extract of Dicystis.

[0030] 3) Preparation method of boiling water extract of Dicystis: Add deionized water or buffer to algal mud or algal powder, stir evenly, boil for 3 min-3 h, let stand and cool to room temperature, centrifuge and collect the supernatant, add an equal amount of deionized water or buffer to algal body and boil for extraction, repeat the above operation 1-30 times, combine the supernatant, freeze dry to obtain the boiling water extract of Dicystis.

[0031] 4) Preparation method of ethanol extract of Dicystis: Add ethanol aqueous solution (volume fraction of 30%-100%) to algal mud or algal powder, stir evenly, and extract with ultrasonic assistance for 1 min-1 h, followed by static cold soaking for 5 min-72 h. Centrifuge and collect the supernatant. Add an equal amount of ethanol aqueous solution to the algal body for further soaking. Repeat the above operation 1-30 times. Combine the supernatants and collect them by rotary evaporation at 30-45℃. This is the ethanol extract of Dicystis.

[0032] 5) Preparation method of bicystic algae ester extract: Add ethyl acetate to algal mud or algal powder, stir evenly, and extract with ultrasonic assistance for 1 min-1 h. Then, let stand and cold soak for 5 min-72 h. Centrifuge and collect the supernatant. Add an equal amount of ethyl acetate to the algae for further extraction. Repeat the above operation 1-30 times. Combine the supernatants and collect them by rotary evaporation at 25-40℃. This is the bicystic algae ester extract.

[0033] Through the above-mentioned three-phase independent extraction process of "water-alcohol-ester", extracts rich in different polar active ingredients can be obtained, achieving comprehensive extraction of hydrophilic and lipophilic functional components.

[0034] Furthermore, the amount of extraction solvent added is 1-50 mL / g algal mud, or 10-500 mL / g lyophilized algal powder.

[0035] Furthermore, the Dicystis extract possesses antioxidant, whitening, anti-inflammatory, and UV-shielding activities.

[0036] This invention also provides the application of *Dystrophariae* extract in the preparation of cosmetics or daily chemical products with antioxidant, whitening, anti-inflammatory or ultraviolet shielding effects.

[0037] Furthermore, the cosmetics or daily chemical products include skin care, washing and care, or makeup products.

[0038] Furthermore, the skincare products include toners, lotions, creams, serums, masks, eye creams, or sunscreens; the personal care products include facial cleansers, shower gels, shampoos, or conditioners.

[0039] The beneficial effects of this invention are as follows:

[0040] (1) Using the intertidal microalgae Dicystis in the South China Sea as raw material, the extract is enriched with polysaccharides, polyphenols, mycospore-like amino acids (MAAs), carotenoids and other active substances through three-phase independent extraction of water-alcohol-ester, so that the extract has antioxidant, whitening, anti-inflammatory and broad-spectrum ultraviolet shielding functions.

[0041] (2) All raw materials are derived from the photobioreactor microalgae cultivation system. The growth cycle is 7-14 days. Each 1.0 kg of dry algae powder produced can fix about 1.8 kg of CO2, achieving negative carbon production. The cultivation process does not require arable land, pesticides, or genetically modified risks.

[0042] (3) The extraction process is simple, energy consumption is low, no toxic organic solvents are used, and it is easy to scale up production, which meets the requirements of green cosmetic raw materials.

[0043] Dicystis ( Geminocystis sp.) MH1 was deposited on January 12, 2026 at the China Center for Type Culture Collection (CCTCC), Wuhan University, Wuhan, China, 430072, China, with accession number CCTCC NO: M2026091. Attached Figure Description

[0044] Figure 1 It is a type of algae called Dicystis. Geminocystis Morphological characteristics of sp. MH1.

[0045] Figure 2 It is a type of algae called Dicystis. Geminocystis Homology alignment analysis of the 16S rDNA sequence of sp. MH1.

[0046] Figure 3 It is a type of algae called Dicystis. Geminocystis Phylogenetic tree of the 16S rDNA gene of sp. MH1. Detailed Implementation

[0047] The following embodiments are further illustrations of the present invention, but not limitations thereof. Specific experimental conditions and methods are not specified in the following embodiments, and the techniques used are generally conventional methods well known to those skilled in the art.

[0048] Example 1: Isolation and identification of Dicystis MH1

[0049] 1. Isolation of Dicystis MH1

[0050] On February 24, 2022, 10L of seawater was collected from Tangjiawan Beach (N 22°20'9'', E 113°35'32'') in Xiangzhou District, Zhuhai City, Guangdong Province. After initial filtration through a 300-mesh filter to remove sediment, the seawater was then filtered through a 0.22μm filter membrane to enrich microalgae. The enriched microalgae on the filter membrane was scraped off and transferred to F / 2 medium for cultivation for approximately 20 days. Observation under an optical microscope showed... Figure 1 As shown, the thallus exists singly or in small, free aggregates, without a distinct gelatinous sheath. The cells are spherical or nearly spherical, becoming hemispherical immediately after division, ranging in color from yellowish-brown to reddish-brown, with uniform intracellular contents and no air sacs. The cells are 2.8-4.9 μm in diameter and reproduce by binary fission.

[0051] 2. Identification of Dicystis MH1

[0052] The algal strain was entrusted to the Freshwater Algae Bank of the Chinese Academy of Sciences for morphological and molecular biological identification. The 16S rDNA fragment sequence of the obtained sample was compared with the GenBank database using BLAST. Sequences of similar algae were downloaded from GenBank, and gene sequences were aligned using Clustal X, supplemented by manual correction using SEAVIEW. A phylogenetic tree was constructed using MEGA5 to analyze the phylogenetic position of the sample.

[0053] Sequencing revealed that the 16S rDNA fragment of this algal strain was 1716 bp in size. Sequence homology comparison with data from the NCBI Genbank database showed that this algal strain was related to the genus *Dicystis* (…). Geminocystis They are closely related, with sequence homology exceeding 99%. Figure 2 ).

[0054] Phylogenetic tree constructed based on 16S rDNA sequence of cell ribosomal ribosomes ( Figure 3 The results showed that the algal strain Contig 1 was related to the genus Dicystis (…). Geminocystis They are closely related and belong to the same evolutionary branch.

[0055] Based on a comprehensive analysis of microscopic observation and 16S rDNA sequence sequencing results, the sample was identified as a species of *Dicystis*. Geminocystis sp., belonging to the phylum Cyanophyta, class Cyanophyceae, order Chroococcales, family Geminoccystaceae, genus Dicystus ( Geminocystis After comparing this algal species with those already reported in the literature of the genus *Dicystis*, it was determined to be a new species of the genus *Dicystis*, and was named *Dicystis*. GeminocystisThe algal strain MH1 was deposited on January 12, 2026 at the China Center for Type Culture Collection (CCTCC), located at Wuhan University, Wuhan, China, 430072, with accession number CCTCCNO: M 2026091.

[0056] Example 2: Preparation of Dicystis extract 1

[0057] Preparation of ultrasonic water extract of Dicystis: Take 5.0g of Dicystis algal mud, add 250mL of PBS buffer, stir evenly, and extract cells by ultrasonic disruption for 3h with power set to 5W. Centrifuge to collect the supernatant, freeze dry, and obtain 0.1g of ultrasonic water extract of Dicystis.

[0058] Example 3: Preparation of Dicystis extract 2

[0059] Preparation of ultrasonic water extract of Dicystis: Take 1.0 g of Dicystis algal powder, add 30 mL of deionized water, stir evenly, and extract cells by ultrasonic disruption for 3 min with the power set to 650 W. Centrifuge and collect the supernatant. Add 30 mL of deionized water to the algal body and continue ultrasonic disruption and extraction. Repeat the above operation 7 times. After freeze drying, 0.46 g of ultrasonic water extract of Dicystis was obtained.

[0060] Example 4: Preparation of Dicystis extract 3

[0061] Preparation of Dicystis freeze-thaw water extract: Take 2.0 g of Dicystis algal powder, add 20 mL of PBS buffer, stir well, and freeze in a -80℃ freezer. After freezing is complete, take it out and thaw, centrifuge and collect the supernatant. Add 20 mL of PBS buffer to the algal body for freeze-thaw extraction. Repeat the above operation 30 times, combine the supernatants, freeze dry, and obtain 0.8 g of Dicystis freeze-thaw water extract.

[0062] Example 5: Preparation of Dicystis aeruginosa extract 4

[0063] Preparation of the boiled water extract of Dicystis: Take 20.0g of Dicystis mud, add 20mL of deionized water, boil and continue heating for 3h for extraction. After cooling the extract to room temperature, centrifuge to collect the supernatant, freeze dry to obtain 0.2g of boiled water extract of Dicystis.

[0064] Example 6: Preparation of Dicystis extract 5

[0065] Preparation of the boiled water extract of Dicystis: Take 1.0 g of Dicystis algal powder, add 500 mL of PBS buffer, boil and continue heating for 3 min for extraction. After cooling the extract to room temperature, centrifuge and collect the supernatant. Add 500 mL of PBS buffer to the algal body and boil for extraction. Repeat the above operation 5 times. Combine the supernatants and freeze dry to obtain 0.4 g of boiled water extract of Dicystis.

[0066] Example 7: Preparation of Dicystis extract 6

[0067] Preparation of the ethanol extract of Dicystis: Take 1.0 g of Dicystis algae powder, add 500 mL of 30% ethanol aqueous solution, extract with ultrasonic assistance for 1 min, then extract with cold soaking for 5 min, collect the supernatant by centrifugation, add 500 mL of 30% ethanol aqueous solution to the algae, repeat the above extraction operation 3 times, combine the supernatants, and collect 0.1 g of Dicystis ethanol extract by rotary evaporation.

[0068] Example 8: Preparation of Dicystis extract 7

[0069] Preparation of ethanol extract of Dicystis: Take 50.0g of Dicystis mud, add 500mL of anhydrous ethanol, extract with ultrasonic assistance for 1h, then extract with cold soaking for 72h, collect the supernatant by centrifugation, add 500mL of anhydrous ethanol to the algae, repeat the above extraction operation 5 times, combine the supernatants, and collect 1.4g of ethanol extract of Dicystis by rotary evaporation.

[0070] Example 9: Preparation of Dicystis extract 8

[0071] Preparation of Bicystis ester extract: Take 5.0 g of Bicystis powder, add 200 mL of ethyl acetate, extract with ultrasonic assistance for 1 h, then extract with cold soaking for 72 h, collect the supernatant by centrifugation, add 500 mL of ethyl acetate to the algae, repeat the above extraction operation 3 times, combine the supernatants, and collect 0.15 g of Bicystis ester extract by rotary evaporation.

[0072] Test Example 1: Test of ABTS's Free Radical Scavenging Ability

[0073] Dissolve trolox (6-hydroxy-2,5,7,8-tetramethyltryptane-2-carboxylic acid) or *Dicystis pilosa* extract in a solvent to prepare sample solutions. Prepare separate solutions of 7 mM ABTS (2,2'-azido-bis-3-ethylbenzothiazoline-6-sulfonic acid) and 2.45 mM K₂S₂O₈ at a 1:1 volume ratio and react at room temperature in the dark for 12-16 h to prepare ABTS solutions. + Working solution. Take 20 μL of sample and add 180 μL of ABTS. + The working solution was reacted at room temperature in the dark for 6 min / 30 min, and the absorbance at 734 nm was measured and recorded as A1; 20 μL of sample was taken, 180 μL of blank solvent was added, and the reaction was carried out at room temperature in the dark for 6 min / 30 min, and the absorbance at 734 nm was measured and recorded as A2; 20 μL of blank solvent was taken, 180 μL of ABTS was added +The working solution was reacted at room temperature in the dark for 6 min and 30 min, and its absorbance at 734 nm was measured and recorded as A0. The ABTS radical scavenging rate was calculated according to the following formula: Scavenging rate = Plotting trolox concentration on the x-axis and ABTS radical scavenging rate on the y-axis, we obtain c. trolox - ABTS free radical scavenging rate standard curve. Calculate the ABTS free radical scavenging rate of the sample and substitute it into the standard curve to obtain the trolox-equivalent antioxidant capacity (TEAC) of the Dicystis extract.

[0074] Test Example 2: Test of Ferrous Reduction Ability (FRAP)

[0075] Dissolve trolox or Diplocystis aeruginosa extract in a solvent to prepare sample solutions. Prepare 0.3 M acetate buffer (pH 3.6), 10 mM TPTZ (dissolved in 40 mM HCl), and 20 mM FeCl3 separately. Mix the three in a 10:1:1 volume ratio and preheat at 37°C for 10 min to prepare FRAP working solutions. Take 20 μL of sample, add 180 μL of FRAP working solution, incubate at 37°C for 4 min, and measure the absorbance at 593 nm, denoted as A1; take 20 μL of sample, add 180 μL of blank solvent, incubate at 37°C for 4 min, and measure the absorbance at 593 nm, denoted as A2. Plot c with trolox concentration on the x-axis and ΔA on the y-axis (ΔA = A2 - A1). trolox -A 593 Standard curve. By substituting the sample's ΔA into the standard curve, the iron-reducing capacity of the *Dicystis jirovecii* extract can be determined.

[0076] Test Example 3: Test for Tyrosinase Inhibitory Activity

[0077] Take kojic acid or Diplocystis extract, dissolve it in a solvent, and prepare a sample solution. Take 57 μL of the above sample, add 29 μL of tyrosinase (10 U / mL, dissolved in citrate buffer), incubate at 37℃ for 10 min, then add 114 μL of 1 mg / mL levodopa, and measure the absorbance at 475 nm, recorded as A1; take 57 μL of the sample, add 29 μL of citrate buffer, incubate at 37℃ for 10 min, then add 114 μL of 1 mg / mL levodopa, and measure the absorbance at 475 nm, recorded as A2; take 57 μL of citrate buffer, add 29 μL of tyrosinase, incubate at 37℃ for 10 min, then add 114 μL of 1 mg / mL levodopa, and measure the absorbance at 475 nm, recorded as A3; take 86 μL of citrate buffer, incubate at 37℃ for 10 min, then add 114 μL of 1 mg / mL levodopa, and measure the absorbance at 475 nm, recorded as A4. Calculate the tyrosinase inhibition rate using the following formula: Inhibition rate = Kojic acid (14.25 μg / mL) was used as a positive control.

[0078] Test Example 4: Test of Cyclooxygenase-2 (COX-2) Inhibitory Activity

[0079] Dicystis aeruginosa extract was dissolved in a solvent to prepare a sample solution. The COX-2 inhibitory activity of the samples was tested using the Beyotime S0168 cyclooxygenase-2 (COX-2) inhibitor screening kit.

[0080] Test Example 5: Sun Protection Factor (SPF) Test

[0081] Dissolve the Dicystis aeruginosa extract in a solvent, filter through a 0.22 μm filter membrane, and measure the absorbance of the filtrate in the wavelength range of 290-320 nm using a UV spectrophotometer. Calculate the SPF value of the sample using the following formula: The EE values ​​for each wavelength are as follows: 290 nm - 0.015, 295 nm - 0.0817, 300 nm - 0.2874, 305 nm - 0.3278, 310 nm - 0.1864, 315 nm - 0.0837, and 320 nm - 0.0180.

[0082] Example 10: Testing of the antioxidant, whitening activity and sun protection factor (SPF) of Dicystis aeruginosa extract

[0083] Following the methods described in Test Examples 1-5, the antioxidant, whitening, anti-inflammatory activities and sun protection factor (SPF) of the *Dicystis jirovecii* extracts obtained in Examples 2-9 were determined, and the results are shown in Tables 1-4.

[0084] Table 1. Antioxidant activity of the extracts from *Dicystis* obtained in Examples 2-9

[0085] *TE: Trolox equivalent.

[0086] Table 2. Tyrosinase inhibitory activity in the extracts of *Dicystis jirovecii* obtained in Examples 2-9

[0087] *The concentration of the Diplocystis aeruginosa extract in each example in Table 2 was 2.9 mg / mL. The positive control kojic acid (14.25 μg / mL) showed a tyrosinase inhibition rate of 55.8%.

[0088] Table 3. Sun Protection Factor (SPF) values ​​of the Dicystis aureus extracts obtained in Examples 2-9

[0089] *The concentration of the Diplocystis extract in each example in Table 3 is 2 mg / mL.

[0090] Table 4. COX-2 inhibitory activity in the extracts of *Dicystis jirovecii* obtained in Examples 2-9

[0091] *The concentration of the Dicystis extract in each example in Table 4 was 125 μg / mL, and the COX-2 inhibition rate of the positive control celecoxib (0.04 μg / mL) was 58.8%.

[0092] The above detailed description is a specific description of the embodiments of the present invention. These embodiments are not intended to limit the patent scope of the present invention. All equivalent implementations or modifications that do not depart from the present invention should be included in the patent scope of this case.

Claims

1. Dicystis ( Geminocystis sp.)MH1, characterized in that, The depositary institution is the China Center for Type Culture Collection, and the accession number is CCTCC NO: M 2026091.

2. A method for cultivating Dicystis aeruginosa, characterized in that, Includes the following steps: 1) The Dicystis thunbergii described in claim 1 is cultured in a microalgae photobioreactor, using F / 2 medium, Zarrouk medium, BG11 medium or modified Dicystis thunbergii-specific medium SYSU-Lvsan-1, at a temperature of 20-40℃, a light intensity of 1000-5000 Lux, and for a culture time of 1-30 days. 2) After cultivation, the algal mud is collected by centrifugation after settling for 1-3 days. The algal mud is then freeze-dried to obtain freeze-dried algal powder.

3. The breeding method according to claim 2, characterized in that, The modified Dicystis-specific culture medium SYSU-Lvsan-1 has the following formula: 53.00 mg / L urea and 5.65 mg / L NaH2PO4·H2O.

4. A method for preparing a Dicystis jirovecii extract, characterized in that, Includes the following steps: Add an extraction solvent to algal mud or algal powder, and extract using physical crushing, thermal extraction or organic solvent extraction. Collect the supernatant by centrifugation, and concentrate or freeze-dry to obtain the extract of Dicystis jirovecii. The physical fragmentation method includes ultrasonic fragmentation or repeated freeze-thaw cycles; The thermal extraction method is boiling extraction; The organic solvent extraction method includes alcohol extraction or ester extraction.

5. The preparation method according to claim 4, characterized in that, The ultrasonic disruption uses deionized water or buffer solution as the extraction solvent, with an ultrasonic power of 5-650 W, an extraction time of 3 min-3 h per extraction, and a total of 1-30 extractions. The repeated freeze-thaw cycles used deionized water or buffer solution as the extraction solvent, with freezing temperatures ranging from -20°C to -80°C and freeze-thaw cycles ranging from 1 to 30 times. The boiling extraction uses deionized water or buffer solution as the extraction solvent, and each extraction time is 3 min-3 h, for a total of 1-30 extractions. The alcohol extraction method uses 30%-100% ethanol as the extraction solvent, with each extraction lasting 5 min-72 h, and is repeated 1-30 times. The ester extraction method uses ethyl acetate as the extraction solvent, with each extraction lasting 5 min to 72 h, and is repeated 1 to 30 times.

6. The preparation method according to claim 4, characterized in that, The extraction solvent is added at a rate of 1-50 mL / g of algal mud or 10-500 mL / g of freeze-dried algal powder.

7. The *Dystrophariae* extract prepared by the method according to any one of claims 4-6, characterized in that, The extract of *Dystrophariae* has antioxidant, whitening, anti-inflammatory, and UV-shielding activities.

8. The use of the Dicystis aurea extract according to claim 7 in the preparation of cosmetics or daily chemical products with antioxidant, whitening, anti-inflammatory or ultraviolet shielding effects.

9. The application according to claim 8, characterized in that, The cosmetics or daily chemical products mentioned include skin care, washing and care, or makeup products.

10. The application according to claim 9, characterized in that, The skincare products include toners, lotions, creams, serums, masks, eye creams, or sunscreens; the personal care products include facial cleansers, shower gels, shampoos, or conditioners.

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