Microalgae with high yield of saponin and application thereof

Abstract

The invention discloses microalgae with high yield of saponin and application of the microalgae, and belongs to the technical field of microalgae biology. The 18S rDNA (ribosomal deoxyribonucleic acid) sequence of the Klebsormidium sp. FDA-14 is shown as SEQ ID NO.3, the Klebsormidium sp. FDA-14 is preserved in the China Center for Type Culture Collection, the preservation address is Wuhan University, Wuhan, China, and the preservation number is CCTCC NO: M 20252282. When the strain is cultured in a BG11 culture medium, saponin can be efficiently produced, and the content of saponin reaches up to 99.49 mg / g and is higher than the content of saponin in ginseng. Compared with ginseng, the yield of the methanol extract of the strain is high, the methanol extract and the crude polysaccharide extract are high in oxidation resistance, and the strain can be used as a potent antioxidant. Compared with other saponin production raw materials, the strain is short in culture period, is not influenced by factors such as environment, region and season, and is expected to provide a new efficient saponin production way which is independent of cultivated land, short in production period and high in content.

Description

Technical Field

[0001] This invention belongs to the field of microalgae biotechnology, and particularly relates to a microalgae that produces high levels of saponins and its applications. Background Technology

[0002] Saponins are glycosides of steroids or triterpenoids, widely found in higher plants and also in small amounts in marine organisms such as starfish and sea cucumbers. As a class of natural active ingredients, saponins are the main active components of many medicinal and edible plants (such as ginseng, Panax notoginseng, and licorice), exhibiting significant anti-inflammatory, antibacterial, antioxidant, anticancer, hypoglycemic, and immunomodulatory activities. With the continuous increase in demand for natural and health products, the saponin market is expanding, and its applications in pharmaceuticals, health products, and cosmetics are becoming increasingly widespread, indicating huge market potential.

[0003] Currently, the main source of saponins still relies on extraction from terrestrial plants, such as soapberry trees, ginseng, licorice, and platycodon. However, these plants are mostly perennials, and their growth is easily affected by environment, season, and region. Furthermore, they have low saponin content and high extraction costs. Taking ginseng as an example, the saponin content of ginseng increases year by year with age, reaching approximately 5% in four-year-old ginseng. Traditional saponin production methods face a series of severe challenges, such as unstable raw material supply, long production cycles, and competition for land with grain crops, which restrict the development of the saponin industry. Therefore, developing a new saponin resource that does not rely on arable land, has a short production cycle, is sustainable, and is easy to standardize is crucial for overcoming current industry bottlenecks.

[0004] Microalgae, as a type of photosynthetic autotrophic organism, possess advantages over higher plants, including rapid reproduction, short growth cycles, strong environmental adaptability, and controllable cultivation conditions. As ancestors of terrestrial higher plants, microalgae contain abundant bioactive components, including many similar to those found in medicinal plants, and are considered a green and sustainable source of various bioactive substances. Furthermore, microalgae cultivation is unaffected by environmental, geographical, or seasonal factors, and has a short cultivation cycle, making it a sustainable new form of biomass production. In recent years, a few studies have reported that some single-celled microalgae can synthesize bioactive saponins, but the content is low (4%), and there are currently no patents for the use of microalgae in saponin production. Utilizing microalgae to produce saponins is of great significance for promoting the development of the saponin industry. Obtaining high-yield, high-quality algal strains is a crucial means to achieve the goal of efficient saponin production from microalgae. Summary of the Invention

[0005] One of the objectives of this invention is to provide a high-saponin-producing strain of Kerria ( Klebsormidium sp.) FDA-14, the Kernelia (sp.) KlebsormidiumThe FDA-14 accession number (sp.) is CCTCC NO: M 20252282.

[0006] The second objective of this invention is to provide the aforementioned Kerria ( Klebsormidium The FDA-14 cultivation method includes the following steps: culturing Kerria using BG11 medium (sp.) Klebsormidium (spe.) FDA-14.

[0007] Preferably, the cultivation conditions are 24-26℃, white light, light intensity of 90-110 µmol / m2 / s, and a CO2 / air mixture is introduced, wherein the volume ratio of CO2 to air in the mixture is 1.5-2%.

[0008] More preferably, the culture period is 18 to 22 days.

[0009] More preferably, the culture period is 20 days.

[0010] The third objective of this invention is to provide a solution derived from the aforementioned Kerria ( Klebsormidium The method for extracting saponins according to FDA-14 includes the following steps: take dried algal powder of Kerria japonica, add methanol solvent at a ratio of 1:18~22, sonicate and crush, collect the supernatant by centrifugation, filter through a 0.45 μm filter membrane and concentrate to obtain a methanol extract containing saponins.

[0011] The fourth objective of this invention is to provide the aforementioned Kerria ( Klebsormidium Application of FDA-14 (sp.) in the production of saponins.

[0012] The fifth objective of this invention is to provide the aforementioned Kerria ( Klebsormidium The use of extracts (sp.) FDA-14 in the preparation of antioxidants, wherein the extract is a methanol extract or a crude polysaccharide extract.

[0013] Compared with the prior art, the present invention has the following beneficial effects: (1) This invention isolates, purifies and screens a high-saponin-producing strain of Kerria from ginseng rhizosphere soil. Klebsormidium sp. FDA-14, when cultured in a column photobioreactor on BG11 medium, has a biomass of up to 4.63 g / L.

[0014] (2) The present invention contains Kerneliae Klebsormidium The total saponin content in sp. FDA-14 is much higher than that in ginseng, at 99.49 mg / g, making it a high-quality raw material for efficient saponin production.

[0015] (3) The present invention contains Kerneliae KlebsormidiumWhen extracting total saponins using methanol (spe. FDA-14), the yield of this algal strain's methanol extract was 2.65 times that of ginseng's methanol extract, and the total saponin content in the methanol extract was 45.45% higher than that in ginseng's methanol extract. The total saponin content of this algal strain was 3.86 times that of ginseng. Compared with ginseng, this algal strain has significant advantages in saponin production in terms of both extraction and content.

[0016] (4) The present invention contains Kerria clavatum Klebsormidium The extract of sp. FDA-14, compared with ginseng extract (methanol extract and crude polysaccharide extract), has strong antioxidant properties, specifically manifested in higher DPPH and ABTS scavenging rates, and can be used as a potent antioxidant.

[0017] (5) The present invention contains Kerneliae Klebsormidium sp. FDA-14 is a filamentous eukaryotic microalga. Compared with other saponin production raw materials, it has a short growth cycle and is not affected by environmental, regional, or seasonal factors. It is expected to provide a new and efficient way to produce saponins that does not rely on arable land, has a short production cycle, and produces high content.

[0018] FDA-14 biopreservation instructions for Klebsiella: Preservation institution: China Center for Type Culture Collection; Accession number: CCTCC NO: M20252282; Deposit date: October 21, 2025; Location of collection: Wuhan University, Wuhan, China; Taxonomic nomenclature: Klebsormidium sp. Attached Figure Description

[0019] Figure 1 Kerneliae in Example 1 Klebsormidium Optical microscope image of sp. FDA-14 (×1000).

[0020] Figure 2 Kerneliae in Example 1 Klebsormidium The position of sp. FDA-14 in the phylogenetic tree.

[0021] Figure 3 Kerria in Example 2 Klebsormidium Growth curve of sp. FDA-14 in BG11 medium.

[0022] Figure 4 Kerria in Example 2 Klebsormidium sp. FDA-14 and ginseng methanol extract content (A), methanol extract and total saponin content in ginseng / microalgae (B).

[0023] Figure 5 Kerria in Example 3 Klebsormidium sp. FDA-14 and ginseng methanol extract and crude polysaccharide extract at different concentrations (0.1, 0.5, 1 and 2 mg / mL) showed DPPH scavenging rate (A) and ABTS scavenging rate (B). Detailed Implementation

[0024] Example 1: Kerria Klebsormidium Isolation and identification of FDA-14 I. Sample Collection Soil samples from the top 0–20 cm layer were collected in August 2024 at a ginseng breeding base in Wanliang Town, Fusong County, Baishan City, Jilin Province (127.32086°N, 42.44858°E).

[0025] II. Kernel Klebsormidium Isolation and purification of sp. FDA-14 Separation and purification steps: Weigh 10 g of the collected soil sample and pour it into an Erlenmeyer flask containing 90 mL of sterilized BG11 medium. Seal the mouth of the Erlenmeyer flask with sealing film, then place the Erlenmeyer flask in a shaker and shake to mix for 30 min to obtain a 10-fold dilution (10... -1 Soil suspension; 10 -1 10 mL of soil suspension was added to an Erlenmeyer flask containing 90 mL of sterile culture medium to obtain 10 -2 Soil suspension; repeat this process to obtain 10 -3 and 10 -4 Soil suspension was then placed in an Erlenmeyer flask for enrichment culture under light conditions of 24–26°C, white light, and a light intensity of 30 µmol / m². 2 Shake the conical flask periodically. Examine under an optical microscope. When the algal cell density reaches 3000–5000 cells / mL, the enrichment culture is complete, and separation and purification can proceed.

[0026] The microalgae were isolated and purified using the plate spreading method and the 96-well plate method. 100 µL of the enriched microalgae culture was evenly spread onto solid BG11 medium (containing 1.5% agar). After single colonies grew, they were picked out with sterile toothpicks and transferred to 150 µL of BG11 in a 96-well plate. The 96-well plate was sealed and incubated in a light incubator for 7-10 days. When the liquid in the 96-well plate turned green or brownish-yellow, the algal culture was examined under an inverted microscope. The algal culture free of contaminating algae and bacteria was transferred to an Erlenmeyer flask containing 100 mL of sterile medium for further expansion culture to obtain pure microalgae.

[0027] The formulation of the BG11 medium is as follows: 1.5 g / L NaNO3, 52.4 mg / L K2HPO4·3H2O, 75 mg / L MgSO4·7H2O, 36 mg / L CaCl2·2H2O, 6.6 mg / L C6H8O7·H2O, 6 mg / L C6H 11 FeNO7, 1.1 mg / L Na2EDTA·2H2O, 20 mg / L Na2CO3, 2.86 mg / L H3BO3, 1.86 mg / L MnCl2·4H2O, 0.22 mg / LZnSO4·7H2O, 0.39 mg / L Na2MoO4·2H2O, 0.08 mg / L CuSO4·5H2O, 0.05 mg / L Co(NO3)2·6H2O.

[0028] III. Kernel Klebsormidium FDA-14 certification 1. Kerria Klebsormidium Morphological identification of FDA-14 Kryptospira Klebsormidium Morphological characteristics of sp. FDA-14: Unbranched filamentous structures composed of a single row of cylindrical cells, 6-8 μm wide and 7-11 μm long, with chloroplasts in sheet form. Figure 1 ).

[0029] 2. Kernel Klebsormidium Molecular biological identification of FDA-14 Algal cells in the logarithmic growth phase were collected by centrifugation, and genomic DNA was extracted using a fungal genomic DNA extraction kit. This genomic DNA was then amplified as 18S rDNA. The amplification system consisted of 50 μL of the following: 1.0 μL DNA sample, 1.5 μL upstream primer (10 μM), 1.5 μL downstream primer (10 μM), 1.0 μL Taq polymerase (5 u / μL), 5.0 μL 10×Buffer, 1.0 μL dNTPs (10 Mm), and 39.0 μL ddH2O.

[0030] The PCR amplification program was as follows: 95℃ pre-denaturation for 5 min; 95℃ pre-denaturation for 30 s, 58℃ annealing for 30 s, 72℃ extension for 1.5 min, repeated for 35 cycles; final extension at 72℃ for 7 min. The upstream and downstream primers were GTAGTCATATGCTTGTCTC (NS1, SEQ ID NO.1) and TCCGCAGGTTCACCTACGGA (NS8, SEQ ID NO.2), respectively.

[0031] The PCR amplification products were sequenced by Shanghai Panoson Biotechnology Co., Ltd. The PCR amplification product was a partial sequence of 18S rDNA, 1727 bp in length.

[0032] Based on the sequencing results, the sequences were aligned with homologous sequences found in the GenBank database using MEGA11 software, and a phylogenetic tree was constructed using the neighbor-joining method (see [link to phylogenetic tree]). Figure 2 The results showed that this algal strain was related to... Klebsormidium The query coverage value of sp. BEA1108B (ON652610.1) reached 99%, and the perIdent value was 99.77%. Based on the phylogenetic tree, this algal strain can be confirmed as *Kreephyra*, with the Latin name... Klebsormidium sp., belonging to the phylum Charophytes, class Klebsormidiophyceae, order Klebsormidiales, family Klebsormidiaceae, and genus Klebsormidium.

[0033] Based on morphological and molecular biological identification results, the algal strain screened in this invention is *Kreecia*. Klebsormidium sp., and named it Kernel. Klebsormidium sp.FDA-14, whose 18S rDNA sequence is shown in SEQ ID NO.3, was deposited at the China Center for Type Culture Collection on October 21, 2025, with accession number CCTCCNO: M 20252282.

[0034] SEQ ID NO.3: agggcgcatg tctagtataa ctcttttata ctgtgaaact gcgaatggct cattaaatca gttatagttt atttgatggt accttactac tcggataacc gtagtaattc tagagctaat acgtgcacca aatcccgact tctggaaggg acgtatttat tagataaaag gccaatgcgg gcttgcccgg tattgcggtg aatcatgata actcgtcgaa tcgcacggcc tttgcgctgg cgatgtttca ttcaaatttc tgccctatca actttcgatg gtaggataga ggcctacat ggtggtaacg ggtgacggag aattagggtt cgattccgga gagggagcct gagaaacggc taccacatcc aaagaaggca gcaggcgcgc aaattaccca atcctgatac agggaggtag tgacaataaa taacaatgct gggctttca aagtctggca attggaatga gtgcaatcta aatccctcaa cgaggatcca ttggagggca agtctggtgc cagcagccgc ggtaattcca gctccaatag cgtatatta agttgttgca gttagaaagc tcgtagttgg attttgggtt ggggcggccg gtccgcctca cggtgtgcac cggccgaccc atccttcttg ccggggacgc gctcctggcc ttaactggtc gggacgtgga gtcggcgatg ttactttgaa aaaattagag tgttcaaagc aggcctacgc tctgaataca ttagcatgga ataacgtgat aggactctgg tcctattgtg ttggtcttcg ggaccggagt aatgattaat agggacagtt ggggatattc gtatttcatt gtcagaggtg aaattcttgg atttatgaaa gacgaacttc tgcgaaagca tttatcaagg atgttttcat taatcaagaa cgaaagttgg gggctcgaag acgatcagat accgtcctag tctcaaccat aaacgatgcc gactagggat tggcggatgt taatttgatg actccgccag caccttatga gaaatcaaag tttttgggtt ccggggggag tatggtcgca aggctgaaac ttaaaggaat tgacggaagg gcaccaccag gagtggagcc tgcggcttaa tttgactcaa cacggggaaa cttaccaggt ccagacatag taaggattga cagattgaga gctctttctt gattctatgg gtggtggtgc atggccgttc ttagttggtg gagtgatttg tctggttaat tccgataacg aacgagacct cagcctgcta actagttaca cgaagattct tctccgtggc caacttctta gagggactat ttggcgtcta cagccaatgg aagtttgagg caataacagg tctgtgatgc ccttagatgt tctgggccgc acgcgcgcta cactgatgaa ttcaacgagt ttataacctg ggccgaaagg tctgggtaat cttgtgaaat ttcatcgtga tggggataga ttattgcaat tattaatctt caacgaggaa ttcctagtaa gcgcgagtca tcagctcgcg ttgattacgt ccctgccctt tgtacacacc gcccgtcgct cctaccgatt gaatg.

[0035] Example 2: Kerria Klebsormidium culture, extract extraction and saponin content of FDA-14. I. Kerria Klebsormidium culture of FDA-14 Kerria cultured to the logarithmic growth phase Klebsormidiumsp. FDA-14 was inoculated at an initial density of 0.06 g / L into a columnar photobioreactor (5.5 cm in diameter) containing 650 mL of BG11 medium, and kept at a constant room temperature of 25°C and a light intensity of 100 µmol / m². 2 Under white light, the microalgae were continuously cultured with a CO2 / air mixture (2%, v / v). Once the microalgae reached a stable growth stage, they were harvested by centrifugation, washed 2-3 times with deionized water, and then freeze-dried to obtain dry algae powder.

[0036] II. Kernel Klebsormidium sp. FDA-14 methanol extract Using ginseng as a control group, 4-year-old ginseng was harvested from a ginseng cultivation base in Wanliang Town, Fusong County, Baishan City, Jilin Province (127.32086°N, 42.44858°E). The ginseng rhizomes were washed, freeze-dried, and ground to obtain ginseng powder. 1.0 g of algae powder or 1.0 g of ginseng powder was added to methanol solvent at a mass ratio of 1:20. The mixture was ultrasonically disrupted at 90% power for 90 min, centrifuged at 8000 ×g for 10 min, and the supernatant was collected. After filtration through a 0.45 μm filter membrane, the supernatant was concentrated by nitrogen blowing to obtain a solid methanol extract. After drying, the extract was stored at -80℃ and then freeze-dried under vacuum. The proportion of the methanol extract to the stem cells could be calculated by weighing.

[0037] III. Kernel Klebsormidium SP. FDA-14 Extraction and Content Determination Using ginseng saponin content as a control group, saponins in microalgae and ginseng were extracted using an ultrasound-assisted methanol method, the same as the methanol extract method. The total saponin content in microalgae and ginseng was determined using the vanillin-perchloric acid colorimetric method. A certain volume of methanol solution was added to the above-mentioned solid methanol extracts of microalgae and ginseng to prepare a 10 mg / mL methanol extract solution. Using ginsenoside monomer Re as a standard, standard solutions of 4 mg / mL, 2 mg / mL, 1 mg / mL, 0.5 mg / mL, 0.1 mg / mL, and 0.05 mg / mL were prepared with methanol. 200 µL of the methanol extract or standard solution was pipetted into 10 mL test tubes, and 2 mL of methanol was added to each tube. 0.5 mL of 5% vanillin-perchloric acid solution was added to the test tubes, and the mixture was shaken well and placed in a water bath at 60°C for 15 min, followed by cooling in ice water. 5 mL of 77% sulfuric acid solution was added to the test tubes, and the absorbance at 540 nm was measured using a UV spectrophotometer. Finally, the total saponin content in the methanol extracts of microalgae and ginseng was calculated based on the standard curve. The total saponin content in microalgae and ginseng was calculated based on the content of the methanol extract.

[0038] Figure 3The results showed that after 20 days of cultivation, Klebsormidium The biomass concentration of sp. FDA-14 is as high as 4.63 g / L, and the biomass productivity is 0.23 g / L / d. Figure 4 The results showed that under these culture conditions, the content of the microalgae methanol extract was 35.44% DM, which was 2.65 times that of the ginseng methanol extract (13.37% DM). The total saponin content in the microalgae methanol extract was as high as 280.66 mg / g, while the total saponin content in the ginseng methanol extract was only 192.97 mg / g. Based on the methanol extract content, the total saponin content in the microalgae could be calculated to be 99.49 mg / g, far exceeding the total saponin content in ginseng (25.79 mg / g), and is 3.85 times that of ginseng. Therefore, Klebsormidium sp. FDA-14 is an excellent algal strain for efficient saponin production, with significantly higher content of methanol extract and total saponins than ginseng.

[0039] Example 3: Kerria Klebsormidium Study on the antioxidant properties of FDA-14 extracts I. Kerria Klebsormidium extraction of FDA-14 and ginseng extract. The microalgae and ginseng extracts include methanol extracts and crude polysaccharide extracts. The extraction method for the methanol extract is the same as in Example 2.

[0040] The crude polysaccharide extraction method is as follows: Take 1.0 g of algae powder or ginseng powder and mix it with deionized water at a mass ratio of 1:25. Heat the mixture in a water bath at 100℃ for 4 h. After the liquid cools down, centrifuge at 8000 g for 10 min to collect the supernatant. Filter the supernatant through a 0.45 μm filter membrane and freeze-dry it to reduce the volume of the supernatant to 1 / 4 of the original volume. Add four times the volume of anhydrous ethanol to the concentrated supernatant, shake to mix, let it stand at 4℃ for 18 h, centrifuge to collect the precipitate, add deionized water to dissolve the precipitate, and freeze-dry to obtain crude polysaccharide powder.

[0041] II. Kernel Klebsormidium Antioxidant properties determination of FDA-14 and ginseng extracts Methanol extracts of microalgae and ginseng were prepared to concentrations of 0.1, 0.5, 1.0, and 2.0 mg / mL with methanol, respectively. Crude polysaccharides were prepared to concentrations of 0.1, 0.5, 1.0, and 2.0 mg / mL with deionized water, respectively. The antioxidant properties of the microalgae and ginseng extracts were determined using DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) and ABTS (2,2'-azido-bis-3-ethylbenzothiazoline-6-sulfonic acid) free radical scavenging rates.

[0042] 1. DPPH free radical scavenging rate DPPH powder was dissolved in anhydrous ethanol to prepare a 48 mg / L DPPH solution, which was stored at -20°C in the dark. 2.0 mL of DPPH solution was mixed with 1.0 mL of sample solution, and the mixture was reacted in the dark for 30 min. The absorbance (A1) was measured at 517 nm. Anhydrous ethanol was used instead of the sample solution, and the absorbance (A0) was measured under the same conditions. Anhydrous ethanol was used instead of the DPPH solution, and the absorbance (A2) was measured under the same conditions. VC was used as a positive control, and each sample was tested in triplicate. The DPPH radical scavenging rate was calculated using the formula: .

[0043] 2. ABTS free radical scavenging rate A 7 mmol / L ABTS solution and a 2.45 mmol / L potassium persulfate solution were mixed at a volume ratio of 1:1 and incubated at room temperature in the dark for 12 h to obtain an ABTS+ radical solution. Before use, the solution was diluted with anhydrous ethanol to achieve an absorbance of 0.70 ± 0.02 at 734 nm. 1 mL of the test sample and 3 mL of the ABTS+ solution were mixed and reacted at 25°C in the dark for 30 min. The absorbance at 734 nm was measured (A1). Anhydrous ethanol was used as a blank control, and the absorbance (A0) was measured. Vitamin C was used as a positive control, and each sample was tested in triplicate. The ABTS radical scavenging rate was calculated using the formula: .

[0044] Antioxidant properties of FDA-14 and ginseng methanol extract and crude polysaccharides, such as Figure 5 As shown, the methanol extract and crude polysaccharide extract of FDA-14, at different concentrations, exhibited higher DPPH and ABTS free radical scavenging rates than ginseng extract, demonstrating better antioxidant properties. When the concentration of crude FDA-14 extract was between 0.5 and 1 mg / mL, it showed excellent scavenging effects on DPPH and ABTS. For the methanol extract, at a concentration of 1 mg / mL, its DPPH scavenging rate was similar to that of vitamin C, reaching 96.25%, which is 4.11 times that of ginseng; at a concentration of 0.5 mg / mL, its ABTS scavenging rate reached 97.51%, which is 2.41 times that of ginseng. For the crude polysaccharide extract, at a concentration of 1 mg / mL, its DPPH scavenging rate was 92.82%, which is 15.00 times that of ginseng; at a concentration of 0.5 mg / mL, its ABTS scavenging rate was the highest (97.87%), which is 6.49 times that of ginseng.

[0045] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A high-saponin-producing strain of Kerria ( Klebsormidium sp.) FDA-14, characterized in that, The Kerria ( Klebsormidium The FDA-14 accession number (sp.) is CCTCC NO: M 20252282.

2. A type of Kerria as described in claim 1 ( Klebsormidium The culture method for FDA-14 (sp.) is characterized by, Includes the following steps: Culturing Kerria using BG11 medium ( Klebsormidium (spe.) FDA-14.

3. The cultivation method according to claim 2, characterized in that, The cultivation conditions are 24–26℃, white light, and a light intensity of 90–110 µmol / m². 2 / s, a CO2 / air mixture is introduced, wherein the volume ratio of CO2 to air in the mixture is 1.5 to 2%.

4. The cultivation method according to claim 2 or 3, characterized in that, The culture period is 18-22 days.

5. The cultivation method according to claim 4, characterized in that, The culture period is 20 days.

6. A method for processing Kerria as described in claim 1 ( Klebsormidium The method for extracting saponins according to FDA-14 (sp.) is characterized by, Includes the following steps: Take dried algal powder of Kerria, add methanol solvent at a ratio of 1:18~22, sonicate to break it up, collect the supernatant by centrifugation, filter it through a 0.45 μm filter membrane and concentrate it to obtain a methanol extract containing saponins.

7. The Kernelosae as described in claim 1 ( Klebsormidium Application of FDA-14 (sp.) in the production of saponins.

8. The Kerneliae as described in claim 1 ( Klebsormidium The application of extracts from FDA-14 (sp.) in the preparation of antioxidants, characterized in that, The extract is a methanol extract or a crude polysaccharide extract.

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