CN114774282B - Critical algae for co-producing gamma-aminobutyric acid and asiatic acid and application thereof - Google Patents

Critical algae for co-producing gamma-aminobutyric acid and asiatic acid and application thereof Download PDF

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CN114774282B
CN114774282B CN202210420496.2A CN202210420496A CN114774282B CN 114774282 B CN114774282 B CN 114774282B CN 202210420496 A CN202210420496 A CN 202210420496A CN 114774282 B CN114774282 B CN 114774282B
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scsio
algae
acid
culture
gaba
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CN114774282A (en
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吴华莲
向文洲
李涛
吴嘉仪
吴后波
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South China Sea Institute of Oceanology of CAS
Southern Marine Science and Engineering Guangdong Laboratory Guangzhou
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Southern Marine Science and Engineering Guangdong Laboratory Guangzhou
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Abstract

The invention discloses a kriging algae for co-producing gamma-aminobutyric acid and asiatic acid and application thereof, belonging to the field of microalgae application. Specifically, the invention provides a Klebsoldium (Klebsoldium sp.) SCSIO-46743 with the preservation number: CCTCCNO: m20211537. Compared with other production raw materials for enriching GABA and asiatic acid, the algae strain has short culture period, is not influenced by factors such as environment, region, seasonality and the like, can quickly adjust the production of the raw materials according to the market demands so as to meet the gap of the raw materials, and can be used for producing GABA and asiatic acid and related health care/functional foods, feeds, medicines, chemical materials, cosmetics, biological products for agriculture/forestry and the like.

Description

Critical algae for co-producing gamma-aminobutyric acid and asiatic acid and application thereof
Technical Field
The invention relates to the field of microalgae application, in particular to a kriging algae for co-producing gamma-aminobutyric acid and asiatic acid and application thereof.
Background
Gamma-aminobutyric acid (GABA), also known as 4-aminobutyric acid, is a natural amino acid composed of non-proteins, and is widely found in vertebrates, plants, and microorganisms. GABA is an important central nervous system inhibitory neurotransmitter in mammals, and has been reported to have various physiological activities such as activating glucose metabolism in the brain, promoting acetylcholine synthesis, anticonvulsant, lowering blood pressure, improving brain function, promoting growth hormone secretion, improving lipid metabolism, anxiolytic, preventing epilepsy, preventing skin aging, delaying brain aging, enhancing reproductive function, and the like. Based on the above-mentioned various physiological functions, GABA has been classified as a new resource food in China. In addition, GABA plays an important role in stress-tolerant physiology of plants and regulation thereof. Therefore, the GABA has wide prospect in the fields of medicines, foods, health products, feeds, cosmetics, biological products for agriculture and forestry and the like. There are 3 methods for GABA production, including chemical synthesis, plant enrichment and microbial metabolism. The chemical synthesis method has the advantages of more synthesis byproducts and poor product safety because of the harsh reaction conditions, so that the product prepared by the method is limited in the application field. Currently, most GABA products are produced by microbial metabolism and plant enrichment. The microbial metabolism method is a fermentation method, and the glutamic acid decarboxylase in the microorganism with high biological safety is utilized to catalyze the decarboxylation reaction to produce GABA. The microbial fermentation process is a heterotrophic process and is easily polluted by other microorganisms, so that the method has high requirements on environment and equipment. The plant enrichment method is to separate and extract GABA from plants with higher GABA content, however, most higher plants have lower GABA content, and some microalgae can also enrich GABA, but the types are very limited, and more microalgae with application value for developing GABA need to be deeply excavated.
Asiatic acid, also known as asiatic acid, is a pentacyclic triterpene compound, and has biological activities such as antiinflammatory, wound healing promoting, skin caring, liver protecting, antidepressant, blood sugar reducing, blood lipid reducing, neuroprotection, and antitumor. Asiatic acid is often used in cosmetics for the purpose of antioxidant, whitening, spot-removing, and skin conditioning. Centella asiatica is an active ingredient of certain traditional Chinese medicine plants, mainly derived from traditional Chinese herbal medicine centella asiatica of Umbelliferae, and is found in higher plants such as loquat leaves, broad-leaved syzygium jambolana, holly, potentilla chinensis and the like, but has not been reported in microalgae. At present, reports that raw materials can co-produce GABA and asiatic acid are not yet available.
Microalgae is a photosynthetic microorganism, and has the advantages of high propagation speed, short growth period, strong environmental adaptability, controllable culture conditions and the like compared with higher plants. In addition, microalgae cells contain abundant physiological active substances, and are ideal resources for developing natural products. The microalgae cultivation is not affected by the factors of environment, region, season and the like, and the cultivation period is short, so that the microalgae cultivation is a sustainable raw material resource. The microalgae is used for extracting and preparing GABA and asiatic acid, and has great significance for promoting the development of microalgae cultivation industry and the wide application of microalgae extraction products in industries such as food, health care, cosmetics and the like.
CN103930560a discloses a method for producing 4-aminobutyric acid from algae, wherein red algae Cyanidium caldarium is acidophilic unicellular microalgae, the most suitable growth pH is 2-3, and growth cannot occur above pH 5. In practical application, the single-cell microalgae culture has the problems of high harvesting energy consumption and high raw material cost.
CN110106210a discloses the use of melatonin in promoting production of gamma-aminobutyric acid by haematococcus pluvialis: and adding melatonin mother liquor into haematococcus pluvialis seed liquid, and inducing haematococcus pluvialis cells to accumulate GABA under the compound stress of continuous illumination and melatonin. The method induces haematococcus pluvialis to synthesize and accumulate GABA through a two-step method.
CN109897873a discloses a method for enriching gamma-aminobutyric acid based on haematococcus pluvialis with good taste: the red algae is cultivated by the combination of culture until the late logarithmic phase, and the red algae is diluted and resuspended by fresh BBM culture medium, and the enrichment of GABA is realized under the combined action of salt (NaCl) and high light. The method realizes the induction from the culture of haematococcus pluvialis to GABA synthesis through a two-step method.
CN102578631B discloses the enrichment method of gamma-aminobutyric acid in green algae: firstly, soaking green algae in purified water, then placing the green algae in a sealed container, applying high pressure of 200-400 MPa to treat the green algae for 5-15 min, and airing the green algae at room temperature under a sterile environment until the weight is constant, thereby improving the GABA content of the green algae. The method has high requirements on conditions, and needs to adopt high-pressure technology treatment and a sterile airing environment.
CN106946971a discloses a process for extracting asiatic acid from centella asiatica: the method for extracting asiatic acid comprises the steps of crushing, primary decoloring, crystallizing, secondary decoloring, recrystallizing, extracting, separating, purifying, concentrating, crystallizing, drying and the like, and only relates to an extraction process, and does not relate to the disclosure of new raw materials.
In summary, the prior art shows the following problems:
(1) GABA content of the currently reported microalgae capable of enriching GABA is not high under normal culture conditions, and the GABA content is required to be improved by a two-step method;
(2) The asiatic acid has single source and is mainly derived from Chinese herbal medicine centella asiatica, and no report on the use of microalgae for producing the asiatic acid exists at present;
(3) Most microalgae are unicellular algae, so that the problems of easy invasion of enemy organisms, high harvesting cost and the like are faced in an open large-scale culture process;
(4) At present, reports on related raw materials for co-production of GABA and asiatic acid are not available.
Disclosure of Invention
In order to solve the technical problems, the invention provides a Klebsiella sp SCSIO-46743 strain which can accumulate GABA and asiatic acid simultaneously and can be used as raw materials for producing GABA and asiatic acid.
The first object of the invention is to provide a Klebsiella sp SCSIO-46743 strain with a preservation number of CCTCC NO: m20211537.
The kringle SCSIO-46743 is obtained by separating and purifying rock samples collected from mountains in Guangzhou urban area.
Morphological characteristics of the Cryptophycoerythrin SCSIO-46743: the unbranched filaments are composed of columnar single-row cells, the cell width is 5-10 μm, the cell length is 8-20 μm, and pigment flakes. The algae can grow in common fresh water algae culture medium.
The second object of the present invention is to provide a preparation comprising the culture of the above-mentioned Klebsiella planticola SCSIO-46743, specifically, the preparation comprising the culture of the Klebsiella planticola SCSIO-46743 strain or a metabolite thereof as an active ingredient.
A third object of the present invention is to provide the use of the above-described krypton SCSIO-46743 or the above-described formulation in one or more of the following:
(1) Preparing gamma-aminobutyric acid (GABA);
(2) Preparing asiatic acid;
(3) Preparing algae powder;
(4) Preparing food, feed, medicine, cosmetic or skin care product containing culture of Sargassum SCSIO-46743 and/or GABA and/or asiatic acid as active ingredients;
(5) Protein was prepared.
The food includes, but is not limited to, functional foods, and in particular, the foods can activate glucose metabolism in brain, promote acetylcholine synthesis, resist convulsion, lower blood pressure, improve brain function, promote growth hormone secretion, improve lipid metabolism, resist anxiety, prevent epilepsy, delay brain aging, enhance reproductive function, and the like.
The medicine can be human medicine and agricultural and forestry biological products, and in particular, the medicine can be used for resisting inflammation, promoting wound healing, reducing blood sugar and blood fat, resisting tumors, protecting liver, resisting depression, protecting nerves, regulating and controlling stress-resistant physiology of plants and the like.
The cosmetic or skin care product can prevent skin aging, promote wound healing, etc.
The fourth object of the present invention is to provide a culture method of the above-mentioned kriging algae SCSIO-46743, comprising the following steps: after the culture of the kresophagitis SCSIO-46743 cultivated to the logarithmic phase is settled, the supernatant is removed and inoculated into a fresh culture medium for cultivation.
Preferably, the cultivation further comprises pouring the algae liquid obtained by inoculating into fresh culture medium into a column type photo-bioreactor for cultivation, wherein the cultivation conditions are as follows: the illumination intensity is 180-200 mu mol/m 2 s 1 The temperature is 25+/-2 ℃, and CO is continuously introduced 2 A gas content of 1%.
Preferably, the composition of the medium is as follows: the concentration of sodium nitrate is 0.3-1.5 g/L, dipotassium hydrogen phosphate is 0.04g/L, magnesium sulfate heptahydrate is 0.075g/L, calcium chloride dihydrate is 0.036g/L, citric acid is 0.006g/L, ferric ammonium citrate is 0.006g/L, EDTA disodium is 0.001g/L, trace element A is 51 ml/L, and the balance is deionized water;
the trace element A5 comprises the following components: 2.86g/L boric acid, 1.81g/L manganese chloride dihydrate, 0.222g/L zinc sulfate heptahydrate, 0.079g/L anhydrous copper sulfate, 0.390g/L sodium molybdate dihydrate and 0.049g/L cobalt nitrate hexahydrate.
A fifth object of the invention is to provide the use of microalgae for the synthesis and accumulation of asiatic acid.
Preferably, the microalgae is of the genus Klebsoldium.
Compared with the prior art, the invention has the following beneficial effects:
(1) The Sdiococcus celebris SCSIO-46743 can accumulate GABA and asiatic acid at the same time, and can be used as raw materials for producing GABA and asiatic acid. Culturing in BG11 culture medium with normal sodium nitrate concentration (1.5 g/L) with tube reactor, and GABA content can reach 0.56%. The culture was carried out in a column reactor in BG11 medium with sodium nitrate concentrations of 0.3,0.5,1.5g/L, the relative centella asiatica content being 6.60%,3.69% and 1.15% of the identifiable substances in the metabolite extract, respectively.
(2) Under the non-induction condition, the GABA content of the kresophagitis SCSIO-46743 accounts for 0.56% of the dry weight of cells, and has very obvious content advantage compared with the existing GABA raw material.
(3) The Cryptophycoerythrin SCSIO-46743 is cultured in BG11 culture medium with normal sodium nitrate concentration (1.5 g/L) by adopting a column reactor, and the biomass can reach 3.35g/L.
(4) In the cell of the gram-lining algae SCSIO-46743, the total amount of amino acids is 30.97g/100g, the types of the contained essential amino acids are complete, 9 essential amino acids account for 40% of the total amino acids, and the protein is high-quality.
(5) The kreson SCSIO-46743 is obtained through liquid culture, can grow in a common fresh water algae culture medium, does not occupy fertile land, has strong environmental adaptability, is easy for outdoor amplification culture, and reduces the raw material culture cost.
(6) Compared with other production raw materials of GABA and asiatic acid, the growth cycle of the kringle algae SCSIO-46743 is short, is not influenced by factors such as environment, region, seasonality and the like, and can be used for rapidly making cultivation adjustment according to the requirements of raw material gaps or markets.
Preservation description
The Klebsoldium sp SCSIO-46743 of the invention is preserved in China Center for Type Culture Collection (CCTCC) at the year 2021, 12 and 03, and has the address of China university of Wuhan, and the post code: 430072, deposit number: cctccc NO: m20211537.
Drawings
FIG. 1 is an optical microscope image of the morphology of the algae filaments and cells of the Cryoalgae SCSIO-46743.
FIG. 2 is a phylogenetic tree of Klebsiella SCSIO-46743 based on 18S rDNA sequences.
FIG. 3 shows the change in biomass dry weight of the Klebsiella in SCSIO-46743 cultured at different sodium nitrate concentrations.
FIG. 4 shows the relative asiatic acid content of the Klebsiella planticola SCSIO-46743 cultured in different sodium nitrate concentrations.
Detailed Description
The present invention will be described in further detail below, but is not limited to, embodiments of the invention, in order to better understand the technical solutions of the present invention for those skilled in the art, with reference to the accompanying drawings and the specific examples.
Example 1 isolation and identification of the algal strain Klebsiella SCSIO-46743
The sample is taken from rock on mountain in Guangzhou city, BG11 culture medium is added after the sample is brought back to a laboratory, the single-algae silk is selected under an inverted microscope and placed in a 96-well plate for single-algae silk culture until a water sample presents visual light green, a coating method is adopted under a sterile condition to separate and purify a solid culture medium of the culture after the single-algae silk culture, a flat plate is placed under weak light normal temperature after coating for culturing for 15-30 days, and the separation steps are repeated for a plurality of times until sterile target algae fall is obtained.
BG11 medium:
the deionized water is added with the following components: 1.5g/L of sodium nitrate, 0.04g/L of dipotassium hydrogen phosphate, 0.075g/L of magnesium sulfate heptahydrate, 0.036g/L of calcium chloride dihydrate, 0.006g/L of citric acid, 0.006g/L of ferric ammonium citrate, 0.001g/L of EDTA disodium and 51 ml/L of trace element A.
Trace element A5 composition: 2.86g/L boric acid, 1.81g/L manganese chloride dihydrate, 0.222g/L zinc sulfate heptahydrate, 0.079g/L anhydrous copper sulfate, 0.390g/L sodium molybdate dihydrate and 0.049g/L cobalt nitrate hexahydrate.
Morphological characteristics of Klebsoldium sp SCSIO-46743: the unbranched filaments were composed of columnar single-row cells, cell width 5-10 μm, cell length 8-20 μm, and pigment flakes (FIG. 1).
Picking single algae from a solid flat plate, placing the single algae into a triangular flask containing 20-30mL of BG11 culture medium, placing the triangular flask on an illumination culture rack for illumination culture, centrifugally collecting algae cells when algae filaments grow to a certain concentration, extracting algae strain DNA by using a plant DNA extraction kit (CTAB method), and amplifying related DNA sequences by using an 18S rDNA universal primer in a PCR reaction condition that: pre-denaturation at 94℃for 5min; denaturation at 94℃for 1min 20s, annealing at 52℃for 1min, extension at 72℃for 2min,36 cycles; finally, the extension is carried out for 10min at 72 ℃. The PCR product was sent to Guangzhou day-Hui gene technologies Co., ltd for sequencing to obtain an 18S rRNA gene partial sequence (cttttatactgtgaaactgcgaatggctcattaaatcagttatagtttatttgatggtaccttactactcggataaccgtagtaattctagagctaatacgtgcaccaaatcccgacttctggaagggacgtatttattagataaaaggccaatgcgggcttgcccggtattgcggtgaatcatgataactcgtcgaatcgcacggcctttgcgctggcgatgtttcattcaaatttctgccctatcaactttcgatggtaggatagaggcctaccatggtggtaacgggtgacggagaattagggttcgattccggagagggagcctgagaaacggctaccacatccaaggaaggcagcaggcgcgcaaattacccaatcctgatacagggaggtagtgacaataaataacaatgctgggcttttcaaagtctggcaattggaatgagtgcaatctaaatccctcaacgaggatccattggagggcaagtctggtgccagcagccgcggtaattccagctccaatagcgtatatttaagttgttgcagttaaaaagctcgtagttggattttgggttggggcagccggtccgcctcacggtgtgcaccggctgacccatccttcttgccggggacgcgctcctggccttaactggtcgggacgtggagtcggcgatgttactttgaaaaaattagagtgttcaaagcaggcctacgctctgaatacattagcatggaataacgtgataggactctggtcctattgtgttggtcttcgggaccggagtaatgattaatagggacagttggggatattcgtatttcattgtcagaggtgaaattcttggatttatgaaagacgaacttctgcgaaagcatttatcaaggatgttttcattaatcaagaacgaaagttgggggctcgaagacgatcagataccgtcctagtctcaaccataaacgatgccgactagggattggcggatgttaatttgatgactccgccagcaccttatgagaaatcaaagtttttgggttccggggggagtatggtcgcaaggctgaaacttaaaggaattgacggaagggcaccaccaggagtggagcctgcggcttaatttgactcaacacggggaaacttaccaggtccagacatagtaaggattgacagattgagagctctttcttgattctatgggtggtggtgcatggccgttcttagttggtggagtgatttgtctggttaattccgataacgaacgagacctcagcctgctaactagttacacgaagattcttctccgtggccaacttcttagagggactatttggcgtctacagccaatggaagtttgaggcaataacaggtctgtgatgcccttagatgttctgggccgcacgcgcgctacactgatgaattcaacgagtttataacctgggccgaaaggtctgggtaatcttgtgaaatttcatcgtgatggggatagattattgcaattattaatcttcaacgaggaattcctagtaagcgcgagtcatcagctcgcgttgattacgtccctgccctttgtacacaccgcccgtcgctcctaccgattgaatgatccggtgaagttttcggattgcggctactccggcggtccgc, specifically shown as SEQ ID NO. 1) of the strain, and homology alignment was performed with NCBI database, and the result showed that the strain was 100% similar to Klebsiella (Klebsormidium elegans), and a phylogenetic tree was constructed based on the 18S rRNA gene sequence, and as shown in FIG. 2, it was confirmed that the strain was Klebsiella sp.
18s rDNA universal primer sequence:
18S-F：5'CCAACCTGGTTGATCCTGCCAGTA3'，
18S-R：5'CCTTGTTAACGACTTCACCTTCCTCT3'。
through morphological and molecular taxonomic identification results, the screened algae strain is Klebsoldium sp, which is named as Klebsoldium sp SCSIO-46743 and is preserved in China Center for Type Culture Collection (CCTCC) at the year 2021, and is named as post code: 430072, deposit number: cctccc NO: m20211537.
Example 2 GABA content of the algal strain Klebsiella SCSIO-46743
After the culture of the Klebsiella planticola SCSIO-46743 cultured to the logarithmic phase was allowed to stand for precipitation, the supernatant medium was removed, and the culture was re-inoculated into 1.2LBG-11 medium (the inoculum size was 0.26 g/L), and the algae liquid was poured into a column type photobioreactor having an optical path of 6cm for cultivation. Culture conditions: the illumination intensity is 180-200 mu mol/m 2 s 1 (light intensity ranges detected at different positions beside the tube) at 25+ -2deg.C, and continuously introducing CO 2 A gas content of 1%. After 18 days of cultivation, a biomass concentration of 3.35g/L (FIG. 3) was obtained, and the algal biomass concentration was measured by a dry weight method. The total amount of amino acids under the culture conditions is 30.97%, the specific amino acid types and contents are shown in Table 1, wherein the GABA content is 0.56%, and the yield is 18.76mg/L. In addition to the cystine and tryptophan content assays using the GBT18246-2000 method, other amino acid content assays using the GB5009.124-2016 method. The BG-11 medium composition was as described in example 1.
TABLE 1 amino acid-producing species of Sdiola SCSIO-46743 and content thereof
Example 3 relative asiatic acid content of the algal strain Klizakii SCSIO-46743
After the culture of the Klebsiella planticola SCSIO-46743 cultured to the logarithmic phase was allowed to stand for precipitation, the supernatant medium was removed, and the culture was re-inoculated into 1.2LBG-11 medium (the inoculum size was 0.26 g/L), and the algae liquid was poured into a column type photobioreactor having a diameter of 6cm for cultivation. Culture conditions: the illumination intensity is 180-200 mu mol/m 2 s 1 The temperature is 25+/-2 ℃, and CO is continuously introduced 2 Gas culture at 1%. After 18 days of continuous illumination culture, the biomass concentration is 3.35g/L(FIG. 3), the measurement method was the same as in example 2. Under this culture condition, the cells were cultured for 18 days, and asiatic acid was 1.15% of the identifiable substance in the extract of the metabolite (FIG. 4). The obtained algae cell methanol extract is subjected to liquid chromatography-mass spectrometry (LC-MS) non-targeted metabonomics full identification, and partial metabolites are finally identified, wherein the relative content of asiatic acid is expressed by the ratio of peak area to the sum of peak areas of identifiable metabolites in a sample. The BG-11 medium composition was as described in example 1.
Example 4 relative asiatic acid content of the algal strain Klizakii SCSIO-46743
After the culture of the kresophagitis SCSIO-46743 cultivated to the logarithmic phase is settled, the supernatant medium is removed, and the culture is inoculated into 1.2L of BG-11 medium limited by nitrogen (the inoculum size is 0.26 g/L), and the algae liquid is poured into a column type photo-bioreactor with an optical path of 6cm for cultivation. Culture conditions: the illumination intensity is 180-200 mu mol/m 2 s 1 The method comprises the steps of carrying out a first treatment on the surface of the The temperature is 25+/-2 ℃, and CO is continuously introduced 2 Gas culture at 1%. After 18 days of continuous light culture, the biomass concentration was 3.48g/L (FIG. 3), and the measurement method was the same as in example 2. Under this culture condition, the cells were cultured for 18 days, and asiatic acid contained 3.69% of the identifiable substance in the extract of the metabolite (FIG. 4), and the measurement method was the same as in example 3.
Nitrogen limited BG-11 medium: the sodium nitrate concentration was 0.5g/L and the other components of the medium were as described in example 1.
Example 5 relative asiatic acid content of the algal strain Klizakii SCSIO-46743
After the culture of the Klebsiella planticola SCSIO-46743 cultivated to the logarithmic phase was allowed to stand for precipitation, the supernatant medium was removed, and the culture was re-inoculated into 1.2L of nitrogen-limited BG-11 medium (inoculum size: 0.26 g/L), and the algae solution was poured into a column type photobioreactor with an optical path of 6cm for cultivation. Culture conditions: the illumination intensity is 180-200 mu mol/m 2 s 1 The temperature is 25+/-2 ℃, and CO is continuously introduced 2 Gas culture at 1%. After 18 days of continuous light culture, the biomass concentration was 3.23g/L (FIG. 3), and the measurement method was the same as in example 2. Under the culture condition, the asiatic acid in the cells can be identified in the extract of the metabolites after culturing for 18 days6.60% of the material (FIG. 4), the measurement method was the same as in example 3.
Nitrogen limited BG-11 medium: the sodium nitrate concentration was 0.3g/L and the other components of the medium were as described in example 1.
The above examples illustrate the various embodiments of the invention in detail, but the embodiments of the invention are not limited thereto. The objects of the present invention can be achieved by one of ordinary skill in the art in light of the present disclosure.
It will be apparent to those skilled in the art that various changes and modifications can be made to the present invention without departing from the principles of the invention, and such changes and modifications fall within the scope of the appended claims.
Sequence listing
<110> national academy of sciences of China, south sea and sea research institute
Guangdong Laboratory of Southern Marine Science and Engineering (Guangzhou)
<120> a gram of algae co-producing gamma-aminobutyric acid and asiatic acid and use thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1645
<212> DNA
<213> gram of the plant Miao-46743 (Klebsoldium sp.SCSIO-46743)
<400> 1
cttttatact gtgaaactgc gaatggctca ttaaatcagt tatagtttat ttgatggtac 60
cttactactc ggataaccgt agtaattcta gagctaatac gtgcaccaaa tcccgacttc 120
tggaagggac gtatttatta gataaaaggc caatgcgggc ttgcccggta ttgcggtgaa 180
tcatgataac tcgtcgaatc gcacggcctt tgcgctggcg atgtttcatt caaatttctg 240
ccctatcaac tttcgatggt aggatagagg cctaccatgg tggtaacggg tgacggagaa 300
ttagggttcg attccggaga gggagcctga gaaacggcta ccacatccaa ggaaggcagc 360
aggcgcgcaa attacccaat cctgatacag ggaggtagtg acaataaata acaatgctgg 420
gcttttcaaa gtctggcaat tggaatgagt gcaatctaaa tccctcaacg aggatccatt 480
ggagggcaag tctggtgcca gcagccgcgg taattccagc tccaatagcg tatatttaag 540
ttgttgcagt taaaaagctc gtagttggat tttgggttgg ggcagccggt ccgcctcacg 600
gtgtgcaccg gctgacccat ccttcttgcc ggggacgcgc tcctggcctt aactggtcgg 660
gacgtggagt cggcgatgtt actttgaaaa aattagagtg ttcaaagcag gcctacgctc 720
tgaatacatt agcatggaat aacgtgatag gactctggtc ctattgtgtt ggtcttcggg 780
accggagtaa tgattaatag ggacagttgg ggatattcgt atttcattgt cagaggtgaa 840
attcttggat ttatgaaaga cgaacttctg cgaaagcatt tatcaaggat gttttcatta 900
atcaagaacg aaagttgggg gctcgaagac gatcagatac cgtcctagtc tcaaccataa 960
acgatgccga ctagggattg gcggatgtta atttgatgac tccgccagca ccttatgaga 1020
aatcaaagtt tttgggttcc ggggggagta tggtcgcaag gctgaaactt aaaggaattg 1080
acggaagggc accaccagga gtggagcctg cggcttaatt tgactcaaca cggggaaact 1140
taccaggtcc agacatagta aggattgaca gattgagagc tctttcttga ttctatgggt 1200
ggtggtgcat ggccgttctt agttggtgga gtgatttgtc tggttaattc cgataacgaa 1260
cgagacctca gcctgctaac tagttacacg aagattcttc tccgtggcca acttcttaga 1320
gggactattt ggcgtctaca gccaatggaa gtttgaggca ataacaggtc tgtgatgccc 1380
ttagatgttc tgggccgcac gcgcgctaca ctgatgaatt caacgagttt ataacctggg 1440
ccgaaaggtc tgggtaatct tgtgaaattt catcgtgatg gggatagatt attgcaatta 1500
ttaatcttca acgaggaatt cctagtaagc gcgagtcatc agctcgcgtt gattacgtcc 1560
ctgccctttg tacacaccgc ccgtcgctcc taccgattga atgatccggt gaagttttcg 1620
gattgcggct actccggcgg tccgc 1645

Claims (7)

1. "Keli" algaeKlebsormidiumsp.) SCSIO-46743 with deposit number: cctccc NO: m20211537.
2. A formulation comprising the kresophagitis SCSIO-46743 of claim 1.
3. Use of the krill-SCSIO-46743 of claim 1 or the formulation of claim 2 in one or more of the following:
(1) Preparing gamma-aminobutyric acid (GABA);
(2) Preparing asiatic acid;
(3) Preparing algae powder;
(4) The preparation method comprises the step of preparing the medicine with the culture of the Szechwan algae SCSIO-46743 and/or GABA and/or asiatic acid as active ingredients.
4. A method of culturing the kresophagitis SCSIO-46743 of claim 1, comprising the steps of: after the culture of the kresophagitis SCSIO-46743 cultivated to the logarithmic phase is settled, the supernatant is removed and inoculated into a fresh culture medium for cultivation.
5. The method according to claim 4, wherein the culturing further comprises pouring the algae liquid obtained by inoculating into a fresh medium into a column type photobioreactor for culturing under the following conditions: the illumination intensity is 180-200 mu mol/m 2 s 1 The temperature is 25+/-2 ℃, and CO is continuously introduced 2 A gas content of 1%.
6. The method according to claim 4, wherein the medium comprises the following components: the concentration of sodium nitrate is 0.3-1.5 g/L, dipotassium hydrogen phosphate is 0.04g/L, magnesium sulfate heptahydrate is 0.075g/L, calcium chloride dihydrate is 0.036g/L, citric acid is 0.006g/L, ferric ammonium citrate is 0.006g/L, EDTA disodium is 0.001g/L, trace element A51 ml/L, and the balance is deionized water;
the trace element A5 comprises the following components: boric acid 2.86g/L, manganese chloride dihydrate 1.81g/L, zinc sulfate heptahydrate 0.222g/L, anhydrous copper sulfate 0.079g/L, sodium molybdate dihydrate 0.390g/L, cobalt nitrate hexahydrate 0.049g/L.
7. Use of the kresophagitis SCSIO-46743 according to claim 1 for the synthesis of accumulated asiatic acid.
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US20150017694A1 (en) * 2008-11-06 2015-01-15 Kiverdi, Inc. Engineered CO2-Fixing Chemotrophic Microorganisms Producing Carbon-Based Products and Methods of Using the Same

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CN109414053A (en) * 2016-02-16 2019-03-01 齐沃生物科学股份有限公司 It is supported via the Animal nutrition for giving the supplement from algae
CN106754389A (en) * 2016-12-29 2017-05-31 中国科学院南海海洋研究所 A kind of method for cultivating microalgae
CN110339101A (en) * 2019-06-14 2019-10-18 广东萱嘉医品健康科技有限公司 A kind of fermentation of seaweed liquid magma and the preparation method and application thereof containing γ-aminobutyric acid

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