Fatty acid-producing mortierella new strain and application thereof

Abstract

The invention relates to a filamentous fungus capable of producing fatty acid, in particular to a new fungus Mortierella sp. 2-1-4 capable of producing the fatty acid. A filamentous fungus Mortierella sp. 2-1-4 is obtained through separation, screening and identification from a soil sample in the Tokeson region of Xinjiang, the strain is subjected to a stress resistance experiment and metabolic fermentation to obtain fatty acid, the fatty acid is verified through the characteristics of salt resistance, pH resistance, growth temperature measurement, functional enzyme experiment, fatty acid production capacity and the like, and the fatty acid can be obtained through the method. The Mortierella sp. Has the advantages that the Mortierella sp. Has good salt-resistant and alkali-resistant characteristics, the Mortierella sp. Can produce fatty acid after being fermented, the yield of the produced fatty acid is 1.33 g / L, and the Mortierella sp. 2-1-4 has a good application prospect of being developed into functional food or medicine and can be widely applied to medicine and food industries.

Description

Technical Field

[0001] The invention relates to the technical field of microbial strains and their applications, and specifically to the technical field of application of a new filamentous fungus 2-1-4 and its fermentation liquid. Background Art

[0002] Fatty acids are a type of carboxylic acid composed of an alkyl chain and a single carboxyl group attached to one end. They are the main components of neutral fats, phospholipids and glycolipids and can be widely used in food and medicine. According to the degree of saturation of the hydrocarbon chain, they can be divided into saturated fatty acids and unsaturated fatty acids. Among them, unsaturated fatty acids include linoleic acid (LA), gamma linolenic acid (GLA), arachidonic acid (ARA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), etc. They have many functions such as lowering blood lipids, improving brain function, and lowering blood sugar. They are a type of nutrient that is essential to the human body, but the human body cannot synthesize it by itself and needs to be supplemented by food intake. Therefore, exploring and developing the source of unsaturated fatty acids has important prospects.

[0003] At present, unsaturated fatty acids are mainly derived from plant seeds, marine fish oils, some mammals and microorganisms. Microorganisms are another major source of polyunsaturated fatty acids because of their short growth cycle, simple culture and high content. At present, there are many reports on fatty acid-producing fungi, such as Mortierella alpina and Mortierella lutea, which metabolize a rich variety of fatty acids with high yields, providing new options for the development of unsaturated fatty acid products. In recent years, with the gradual exploration of microbial resources in special environments, they have shown broad application prospects in the production and utilization of fatty acids. Therefore, it is necessary to continuously explore and find better strains or strains with better properties and applications. Summary of the invention

[0004] In view of the widespread demand for unsaturated fatty acids and the need for fatty acid production strains. The present invention aims to provide a high-yield fatty acid filamentous fungus, specifically a high-yield fatty acid filamentous fungus Mortierella and its application. The present invention isolates and screens a high-yield fatty acid filamentous fungus Mortierella ( Mortierella sp. )2-1-4, through optimizing the fatty acid production conditions of the strain, functional enzyme experiments, stress resistance experiments and antibacterial properties, it was verified that the strain has the ability to produce fatty acids, as well as good salt resistance, alkali resistance and antibacterial properties. Mortierella sp.) The fatty acid yield of 2-1-4 is 1.33 g / L, and the fatty acids produced are rich in variety; this indicates that the new bacteria 2-1-4 provided by the present invention has a good application prospect of being developed into functional food or medicine.

[0005] In order to achieve the above technical effects, the present invention is implemented through the following technical solutions.

[0006] The present invention provides a strain of Mortierella ( Mortierella sp .) 2-1-4, the strain 2-1-4 is deposited in China Center for Type Culture Collection (CCTCC), and the strain collection number is CCTCC NO: M 20242346.

[0007] The filamentous fungus Mortierella ( Mortierella sp. ) 2-1-4 was isolated and screened from soil samples in Toksun County, Xinjiang. After phylogenetic analysis and morphological analysis of the ITS gene sequence, strain 2-1-4 belongs to the genus Mortierella. By sequencing the gene of the strain, the obtained sequence was subjected to BLAST comparison analysis on the NCBI website. The ITS gene sequence of strain 2-1-4 was consistent with the standard model bacteria of the genus. Mortierella antarctica The homology of CBS 609.70 T was 94.69%. Sequences with higher homology were selected to construct the phylogenetic tree of ITS gene sequences. Mortierella antarctica CBS 609.70T clustered into one branch. After polyphasic classification, strains 2-1-4 and Mortierella antarctica CBS 609.70 T was significantly different, confirming that the strain was Mortierella ( Mortierella sp. ) 2-1-4 is a new species of the genus Mortierella and has the typical characteristics of new species.

[0008] The strain Mortierella provided by the present invention Mortierella sp. ) 2-1-4 Through the above-mentioned well-known and recognized strain system molecular level identification in the art, combined with morphological identification analysis, the present invention relates to Mortierella ( Mortierella sp. ) 2-1-4 can form white aerial mycelium, which is cotton-like, relatively thick and has septa. After 9 days of cultivation, the colony diameter reaches 6.8-7.2 cm.

[0009] By using the above provided fungus species Mortierella Mortierella sp. ) 2-1-4, the strain amylase and lipase activities were positive; the growth temperature range was 4~35℃, and it could grow normally in the environment of 5% NaCl and pH 13.0. Through strain comparison, it was different from neighboring strains Mortierella antarctica There are clear differences in CBS 609.70 T.

[0010] The above-mentioned strain identification, through the well-known and recognized strain system molecular level identification and strain morphological verification in the art, confirmed that the obtained strain number 2-1-4 is a typical new strain of Mortierella in the category of Mortierella ( Mortierella sp. )2-1-4, the strain has been deposited in the Budapest Treaty International Microbiological Depository: China Center for Type Culture Collection (CCTCC) before the application date. Address: Room 211, China Center for Type Culture Collection, Wuhan University, Wuchang District, Wuhan City, Hubei Province, Postal Code: 430061, Deposit Number: CCTCC NO: M 20242346, Deposit Date is October 25, 2024.

[0011] The above-mentioned strain Mortierella Mortierella sp. ) The gene sequence of 2-1-4 is shown in SEQ ID NO: 1.

[0012] The strain Mortierella ( Mortierella sp. ) The separation medium of 2-1-4 is: potato glucose agar (containing chloramphenicol) is made of 6.0 g / L potato powder, 20.0 g / L glucose, 20.0 g / L agar, 0.1 g / L chloramphenicol, 1000 mL distilled water, pH 6.0-6.5.

[0013] The strain Mortierella ( Mortierella sp. ) The purification medium of 2-1-4 is: potato glucose agar (containing chloramphenicol) is made of 6.0 g / L potato extract powder, 20.0 g / L glucose, 20.0 g / L agar, 0.1 g / L chloramphenicol, 1000 mL distilled water, pH 6.0-6.5.

[0014] The strain Mortierella ( Mortierella sp. ) The fermentation medium of 2-1-4 is OW medium with 50 g / L glucose, 10 g / L yeast extract powder, KH 2 PO 3 3 g / L, Na 2 SO 4 1 g / L, CaCl 2 0.38 g / L, MgCl 2 •6H 2 O0.5g / L, 1000mL distilled water, pH5.5-6.0.

[0015] At the same time, the present invention provides a new fungus Mortierella ( Mortierella sp. ) 2-1-4 Method for preparing fermentation broth, the fermentation broth is prepared by the following steps: washing the activated Mortierella ( Mortierella sp.) spores on the 2-1-4 plate to obtain a spore suspension, inoculate the 2-1-4 spore suspension into the fermentation medium at a volume ratio of 5%, and ferment and culture at 25°C and 150 rpm for 7 days to obtain the fermentation broth of strain 2-1-4.

[0016] Furthermore, the present invention provides a new filamentous fungus Mortierella ( Mortierella sp. )2-1-4 Application in the preparation of functional foods or medicines.

[0017] Furthermore, the present application also provides a method for preparing fatty acids, using a new strain of high-yield fatty acid fungus Mortierella ( Mortierella sp. )2-1-4 prepared by fermentation.

[0018] The method for preparing a fatty acid comprises inoculating the spore mold ( Mortierella sp. ) 2-1-4 inoculated with OW medium, placed in a shaker at 15-30℃, 100-200 rpm for fermentation for 4-9 days, The method for preparing a fatty acid adopts any one of lactose, fructose, sucrose, corn flour and maltose as a carbon source during fermentation, and adopts any one of peptone, beef extract powder, ammonium sulfate, urea and nitrite as a nitrogen source during fermentation.

[0019] By implementing the above specific technical solutions provided by the present invention and implementing the content of the present invention, the following beneficial effects can be obtained: (1) The present invention provides a strain of Mortierella sp. ( Mortierella sp. ) 2-1-4, and it is necessary to deposit in accordance with legal requirements. Mortierella sp. ) 2-1-4 was isolated and screened from soil samples in Toksun County, Xinjiang, and a high-yield fatty acid filamentous fungus was obtained. The strain was verified by optimizing the fatty acid production conditions, functional enzyme experiments, stress resistance experiments, and antibacterial properties. The strain has fatty acid capacity, as well as good salt resistance, alkali resistance, and antibacterial properties. The strain has a high fatty acid yield after fermentation, and the types of fatty acids produced are rich; this indicates that the strain 2-1-4 provided by the present invention has good application prospects in the food and pharmaceutical industries.

[0020] (2) The novel filamentous fungus Mortierella ( Mortierella sp. )2-1-4 has the ability to produce fatty acids, the fatty acids produced are rich in variety, and have good salt resistance, alkali resistance and antibacterial properties. Mortierella sp.)2-1-4 after fermentation, the fatty acid yield was 1.33 g / L. After the strain was fermented, the fatty acids produced were diverse, providing a new choice for the development of unsaturated fatty acid drugs, indicating that the filamentous fungus Mortierella ( Mortierella sp. )2-1-4 has good application prospects in being developed into functional food or medicine.

[0021] (3) The new filamentous fungus Mortierella provided by the present invention ( Mortierella sp. ) 2-1-4 has the ability to produce lipase. When the strain culture temperature is 25°C and the buffer pH value is 6.5, the fatty acid production is the largest, the fatty acids produced are rich in variety, and they grow well under low temperature and high pH conditions. They have good starch hydrolysis ability and strong antibacterial ability, and can effectively inhibit certain pathogens such as Fusarium oxysporum. Therefore, the present invention can not only be used as a functional food to supplement the necessary nutrients for the human body, but also can inhibit Fusarium oxysporum; the fatty acids produced by strain 2-1-4 can also be added to food. Compared with the fatty acids produced by some bacteria, the new filamentous fungus Mortierella ( Mortierella sp. ) The fatty acids produced by 2-1-4 are safer and more reliable. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Shown is the filamentous fungus Mortierella neoformans ( Mortierella sp. )2-1-4 Phylogenetic tree constructed based on ITS gene sequence.

[0023] Figure 2 Shown is the strain Mortierella ( Mortierella sp. )2-1-4 morphological characteristics diagram, where A is the colony morphology on PDA culture medium; B is the mycelium morphology under electron microscope; C is the mycelium morphology under stereo microscope.

[0024] Figure 3 Shown is the strain Sporophyceae ( Mortierella sp. )2-1-4 Biomass and fermentation yield after fermentation.

[0025] Figure 4 Shown is the effect of temperature on the strain Sporophycetes ( Mortierella sp. )2-1-4 Influence on the performance of fermentation to produce fatty acids.

[0026] Figure 5 Shown is the effect of time on the strain Sporophyceae ( Mortierella sp. )2-1-4 Influence on the performance of fermentation to produce fatty acids.

[0027] Figure 6 Shown is the effect of inoculum size on the strain Sporophycetes ( Mortierella sp. )2-1-4 Influence on the performance of fermentation to produce fatty acids.

[0028] Figure 7Shown is the effect of carbon source on the strain Sporophycetes ( Mortierella sp. )2-1-4 Influence on the performance of fermentation to produce fatty acids.

[0029] Figure 8 Shown is the effect of nitrogen source on the strain Sporophycetes ( Mortierella sp. )2-1-4 Influence on the performance of fermentation to produce fatty acids.

[0030] Fig. 9 Shown is the effect of rotation speed on the strain Sporophyceae ( Mortierella sp. )2-1-4 Influence on the performance of fermentation to produce fatty acids.

[0031] Fig.10 Shown is the strain Sporophyceae ( Mortierella sp. )2-1-4 Main fatty acid content after fermentation. DETAILED DESCRIPTION

[0032] The present invention is described below with reference to the following examples, but the present invention is not limited to the following examples. All raw and auxiliary materials used in the present invention, as well as the selected bacterial strain culture methods are well known in the art, and the % involved in the present invention is the mass volume percentage, unless otherwise specified.

[0033] In order to better explain the present invention, the main contents of the present invention are further illustrated below in conjunction with specific examples, but the contents of the present invention are not limited to the following examples. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available commodities.

[0034] The potato glucose agar (containing chloramphenicol) of the present invention adopts 6.0 g / L potato powder, 20.0 g / L glucose, 20.0 g / L agar, 0.1 g / L chloramphenicol, 1000 mL distilled water, and pH 6.0-6.5.

[0035] The OW culture medium of the present invention comprises 50 g / L glucose, 10 g / L yeast extract powder, and KH 2 PO 3 3 g / L, Na 2 SO 4 1 g / L, CaCl 2 0.38 g / L, MgCl 2 •6H 2 O 0.5g / L, 1000mL distilled water, pH 5.5-6.0.

[0036] The protein skimmed milk powder culture medium of the present invention adopts 12 g / L skimmed milk powder, 4.0 g / L peptone, 3.0 g / L yeast powder and 20 g / L agar.

[0037] The cellulose plate of the present invention adopts 10 g / L sodium carboxymethyl cellulose, Na 2 CO 3 2 g / L, MgSO 4 0.5 g / L, K 2 HPO 4 10 g / L, FeSO 4 0.01 g / L, KCl 0.5 g / L, agar 20 g / L, and the pH value was adjusted to 7.0-7.4.

[0038] The starch plate of the invention adopts 10 g / L starch, 3 g / L beef extract powder, 10 g / L peptone, 5 g / L NaCl and 20 g / L agar, and the pH value is adjusted to 7.0-7.4.

[0039] The lipase plate of the present invention adopts a potato glucose agar (containing chloramphenicol) solid culture medium added with 1% tributyrin and 0.05% rhodamine B, and the pH value is adjusted to 7.0-7.4.

[0040] Example 1: Isolation, purification and identification of strain 2-1-4 1. Separation and purification The gradient dilution method was used to separate filamentous fungi. Soil samples collected from Toksun, Xinjiang were separated and purified by gradient dilution. 5 g of soil sample was weighed and placed in 45 mL of sterile water. Sterilized glass beads were added and shaken at 150 rpm and 30°C for 30 min to fully disperse the sample. Standard gradient dilution was used to dilute 10 -1 -10 -3 Concentration dilution; take 100 μL of each concentration dilution, spread it on a potato dextrose agar (containing chloramphenicol) medium plate by conventional spreading method, and place it in a 30℃ constant temperature incubator for culture; after one week, when colonies grow on the plate, pick a single colony on the plate for purification culture. The purified single colony is transferred to a potato dextrose agar (containing chloramphenicol) medium slope for storage.

[0041] 2. ITS gene identification 1. Extraction of PCR Template DNA The purified strain 2-1-4 was inoculated into potato glucose liquid medium and cultured in a shaking incubator at 25°C for 7 days. The bacteria were collected and the total genomic DNA was extracted using a fungal genomic DNA extraction kit.

[0042] 2. PCR Amplification ITS gene sequence primers: ITS1: 5'-GGTCATTTAGAGGAAGTAA-3'; ITS4: 5'-AGCCTSSCCTTANTDATATGC-3'; PCR reaction system: 2X PCR Bestaq MasterMix 25 μL, forward and reverse primers (10 μmol / L) 1 μL each, DNA template (20 ng / μL) 2 μL, ddH2O to 30 μL. PCR reaction conditions: 95℃ pre-denaturation for 5 min; 94℃ denaturation for 45 s; 50℃ annealing for 30 s, 72℃ extension for 45 s, 30 cycles; 72℃ 1 min after the cycle.

[0043] 3. Sequence Determination The PCR amplification product was sequenced after electrophoresis detection and purification, and its sequence length was 662bp. The test results are shown in SEQ ID No: 1. BLAST homologous sequence search was performed on NCBI, and the phylogenetic tree was established by the Neighbor-Joining method using MEGA 11.0 software commonly used in the field (repeated sampling 1000 times). The results are shown in the attached Figure 1 As shown, the obtained sequence was compared and analyzed on the NCBI website and it was found that strain 2-1-4 was similar to the standard model bacteria of this genus. Mortierella antarctica The homology of CBS609.70 T is 94.69%. In the phylogenetic tree constructed with ITS gene sequences, the ITS gene sequence of strain 2-1-4 is similar to that of strain Mortierella antarctica CBS 609.70 T clustered into a branch. This indicates that strain 2-1-4 may be related to Mortierella antarctica CBS 609.70 T is close. Through the comprehensive judgment of the similarity and homology of the strains, it is consistent with the conclusion of the typical analysis of the molecular level identification of new strains in the field of strain identification, confirming the obtained Mortierella The species number within the genus is 2-1-4, which is a typical new species.

[0044] Example 2: Physiological and biochemical characteristics of strain 2-1-4 1. Colony morphology of strain 2-1-4 Inoculate the strain 2-1-4 to be observed onto a potato dextrose (containing chloramphenicol) agar plate and culture at 30°C for 9 days. When the colonies have grown all over the plate, observe and record the characteristics of the colonies and take photos for preservation. At the same time, record the electron microscope and stereo microscope photos of the hyphae. The results are recorded in the attached Figure 2 shown.

[0045] By the attached Figure 2The results show that the colonies of the strain 2-1-4 provided by the present invention on the PDA medium are petal-shaped, white, with abundant aerial hyphae, cotton-like, relatively thick, septate, and with a typical odor. After 9 days of cultivation, the colony diameter reaches 6.8-7.2 cm.

[0046] Based on the above biological characteristics, the above strain 2-1-4 was identified as Mortierella sp. This strain has been deposited in the Budapest Treaty International Microbiological Depository: China Type Culture Collection (CCTCC), address: Wuhan University, Wuhan, China, postal code: 430072, deposit date: October 25, 2024, deposit number: CCTCC NO: M20242346.

[0047] 2. Physiological and biochemical characteristics of strain 214L11 Strain 2-1-4 was inoculated on potato dextrose (containing chloramphenicol) agar medium, and the physiological and biochemical characteristics were detected. The growth temperature, salinity, pH range, protease, amylase, lipase, cellulase, antibacterial properties were detected. Strain 2-1-4 can grow at 4-35℃, pH5.0-13.0, and can tolerate 5% NaCl. Its metabolic lipase and amylase activities are both positive. In the antibacterial assay against pathogens, the strain showed a certain resistance to Fusarium.

[0048] The ITS gene sequence homology comparison analysis, morphological identification, physiological and biochemical characteristics comparison and other aspects provided in Example 1 are clearly different from the standard model strain, which is consistent with the typical analysis conclusion of the new strain identification in the field. It is determined that the strain 2-1-4 is a new species of the genus Mortierella in a filamentous fungus, and has the significant characteristics of a new species in the same genus Mortierella.

[0049] Example 3: Fermentation broth of the new strain of Mortierella 2-1-4 In this example, based on Example 1 and Example 2, the fermentation broth of the new strain 2-1-4 of Mortierella was obtained by the following steps: the spores on the activated Mortierella plate were rinsed with sterile water to obtain a spore suspension, the 2-1-4 spore suspension was inoculated into OW culture medium at a volume ratio of 5%, and fermented and cultured at 25° C. and 150 rpm for 7 days to obtain the fermentation broth of strain 2-1-4.

[0050] Example 4: Application of the new strain of Mortierella 2-1-4 after fermentation Based on Examples 1 to 3, this example tests strain 2-1-4 and its application after fermentation.

[0051] 1. Experimental study on the performance of the new strain 2-1-4 of Mortierella to produce fatty acids by fermentation The dried cells of the new strain of Mortierella 2-1-4 after fermentation were used for relevant fatty acid production performance test. By the attached Figure 3 The results show that the new strain 2-1-4 provided by the present invention has a good ability to produce fatty acids. It is calculated that the biomass of the strain 2-1-4 after fermentation is 10.56 g / L, and the fermentation yield of fatty acids is 1.33 g / L.

[0052] 2. Determination of the fermentation conditions for fatty acid production by the new strain of Mortierella 2-1-4 (1) Optimal fermentation temperature for fatty acid production The spore suspension of Mortierella new strain 2-1-4 was inoculated into OW medium at a volume ratio of 5%, and placed in a shaker at 15℃, 20℃, 22℃, 25℃, 28℃, and 30℃, respectively, and fermented at 150 rpm for 7 days. The cells were collected by filtration with four layers of sterile gauze and dried in an oven at 60℃ to constant weight to investigate the effect of temperature change on the fermentation yield of fatty acids of Mortierella new strain 2-1-4. The specific results are shown in the attached Figure 4 shown.

[0053] Depend on Figure 4 The data show that the fatty acid fermentation yield of the new Mortierella fungus 2-1-4 provided by the present invention shows a trend of first increasing and then decreasing with increasing temperature, and the optimal fermentation temperature is 25°C, after which the fatty acid fermentation yield begins to decrease.

[0054] (2) Optimal fermentation time for fatty acid production The spore suspension of Mortierella 2-1-4 was inoculated into OW medium at a volume ratio of 5%, and then placed in a 30℃ shaker at 150 rpm for fermentation for 4d, 5d, 6d, 7d, 8d, and 9d, respectively. The cells were collected by filtration through four layers of sterile gauze and dried in an oven at 60℃ to constant weight to investigate the effect of time changes on the fermentation yield of fatty acids of Mortierella 2-1-4. The specific results are shown in the attached Figure 5 shown.

[0055] Depend on Figure 5 The data show that the fatty acid fermentation yield of the new Mortierella fungus 2-1-4 provided by the present invention first increases and then remains stable with the increase of fermentation time. The optimal fermentation time is 7 days, after which the fatty acid fermentation yield remains stable.

[0056] (3) Optimal fermentation inoculum amount for fatty acid production The spore suspension of Mortierella 2-1-4 was inoculated into OW medium at a volume ratio of 3%, 5%, 7%, 9%, 11%, and 13%, respectively, and placed in a 30℃ shaker and fermented at 150 rpm for 7 days. The cells were collected by filtration through four layers of sterile gauze and dried in an oven at 60℃ to constant weight to investigate the effect of inoculation amount on the fermentation yield of fatty acids of Mortierella 2-1-4. The specific results are shown in the attached Figure 6 shown.

[0057] Depend on Figure 6 The data show that the fatty acid fermentation yield of the new Mortierella fungus 2-1-4 provided by the present invention shows a trend of first increasing and then decreasing with the increase of inoculation amount, and the optimal inoculation amount is 7%, after which the fatty acid fermentation yield begins to decrease.

[0058] (4) Optimal fermentation carbon source for fatty acid production Lactose, fructose, sucrose, corn flour and maltose were used to replace the carbon source in OW medium, and the spore suspension of Mortierella new strain 2-1-4 was inoculated into the medium at a volume ratio of 5%. The medium was placed in a shaker at 30℃ and fermented at 150 rpm for 7 days. The cells were collected by filtration with four layers of sterile gauze and dried in an oven at 60℃ to constant weight. The effects of different carbon sources on the fermentation yield of fatty acids of Mortierella new strain 2-1-4 were investigated. The specific results are shown in the attached Figure 7 shown.

[0059] Depend on Figure 7 The data show that the new strain 2-1-4 of Mortierella provided by the present invention has different fatty acid fermentation yields with different carbon sources. When maltose is used as a carbon source, its fatty acid fermentation yield is the highest. When lactose and fructose are used as carbon sources, there is no significant difference in its fatty acid fermentation yield.

[0060] (5) Optimal nitrogen source for fermentation of fatty acids The nitrogen source in OW medium was replaced by peptone, beef extract powder, ammonium sulfate, urea and nitrite, respectively. The spore suspension of Mortierella new strain 2-1-4 was inoculated into the medium at a volume ratio of 5%, and the medium was placed in a 30℃ shaker and fermented at 150rpm for 7 days. The cells were collected by filtration with four layers of sterile gauze and dried in an oven at 60℃ to constant weight. The effects of different nitrogen sources on the fermentation yield of fatty acids of Mortierella new strain 2-1-4 were investigated. The specific results are shown in the attached Figure 8 shown.

[0061] Depend on Figure 8 The data show that the new strain 2-1-4 of Mortierella provided by the present invention has different fatty acid fermentation yields with different nitrogen sources. When yeast extract powder is used as a nitrogen source, its fatty acid fermentation yield is the highest. When urea and nitrite are used as nitrogen sources, its fatty acid fermentation yield and biomass are zero.

[0062] (6) Optimal fermentation speed for fatty acid production The spore suspension of Mortierella new strain 2-1-4 was inoculated into OW medium at a volume ratio of 5%, and was placed in a shaker at 100rpm, 120rpm, 140rpm, 160rpm, 180rpm and 200rpm for fermentation at 30℃ for 7 days. The bacteria were collected by filtering with four layers of sterile gauze and dried in an oven at 60℃ to constant weight to investigate the effect of speed change on the fermentation yield of fatty acids of Mortierella new strain 2-1-4. The specific results are shown in the attached Fig. 9 shown.

[0063] Depend on Fig. 9 The data show that the fatty acid fermentation yield of the new Mortierella sp. 2-1-4 provided by the present invention shows a trend of first increasing and then decreasing with the increase of the rotation speed, and the optimal rotation speed is 160 rpm, after which the fatty acid fermentation yield begins to decrease.

[0064] (7) Analysis of the main fatty acids produced by the new strain 2-1-4 of Mortierella Gas chromatography was used to investigate the composition and content of the main fatty acids in the new strain of Mortierella 2-1-4. Fig.10 shown.

[0065] Depend on Fig.10 The data show that the new strain 2-1-4 of Mortierella provided by the present invention has significant differences in the content of different fatty acids produced. Oleic acid, palmitic acid, stearic acid and arachidonic acid are the main fatty acids, among which oleic acid has the highest content of 32.58%, followed by palmitic acid at 17.71%.

[0066] The embodiments described above are only some embodiments of the present invention, not all embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but only represents selected embodiments of the present invention. All other embodiments obtained without creative work and related deductions and substitutions made by ordinary technicians in the field under the conditions of the concept of the present invention belong to the scope of protection of the present invention.

Claims

1. A new strain of high-yield fatty acid Mortierella Mortierella sp. ) 2-1-4, characterized in that, New fungus Mortierella Mortierella sp. )2-1-4 is deposited in China Center for Type Culture Collection (CCTCC), with strain accession number CCTCCNO: M 20242346. The spore mold ( Mortierella sp. ) The gene sequence of strain 2-1-4 is shown in SEQ ID NO:

1.

2. Used to isolate a new strain of high-yield fatty acid filamentous fungus Mortierella ( Mortierella sp. ) The culture medium of 2-1-4, characterized in that The new fungus Mortierella Mortierella sp. ) The separation medium of 2-1-4 is: potato glucose agar (containing chloramphenicol) is made of 6.0 g / L potato extract powder, 20.0 g / L glucose, 20.0 g / L agar, 0.1 g / L chloramphenicol, 1000 mL distilled water, pH 6.0-6.

5.

3. Used for purifying a novel strain of Mortierella sp. ( Mortierella sp. ) The culture medium of 2-1-4, characterized in that The purification medium of the 2-1-4 strain is: potato glucose agar (containing chloramphenicol) using 6.0 g / L potato extract powder, 20.0 g / L glucose, 20.0 g / L agar, 0.1 g / L chloramphenicol, 1000 mL distilled water, pH 6.0-6.

5.

4. Used for fermentation culture of a novel high-yield fatty acid fungus Mortierella ( Mortierella sp. ) The culture medium of 2-1-4, characterized in that The fermentation medium of the 2-1-4 strain is OW medium containing: 50 g / L glucose, 10 g / L yeast extract powder, 3 g / L KH2PO3, 1 g / L Na2SO4, 0.38 g / L CaCl2, 0.5 g / L MgCl2•6H2O, 1000 mL distilled water, pH 6.0-6.

5.

5. A novel strain of high-yield fatty acid Mortierella as claimed in claim 1 ( Mortierella sp. ) The fermentation broth of 2-1-4, characterized in that The fermentation broth is prepared by the following steps: rinsing the activated spores on the Mortierella plate with sterile water to obtain a spore suspension, inoculating the 2-1-4 spore suspension into an OW culture medium at a volume ratio of 5%, fermenting and culturing at 25° C. and 150 rpm for 7 days, and obtaining a fermentation broth of strain 2-1-4.

6. The novel high-yield fatty acid bacterium Mortierella as claimed in claim 1 ( Mortierella sp. )2-1-4 Application in the preparation of functional foods or medicines.

7. The method for separating a novel high-yield fatty acid filamentous fungus Mortierella according to claim 1 according to claim 2. Mortierella sp. ) Application of the culture medium of 2-1-4 in the preparation of functional foods or medicines.

8. or the method for purifying a novel strain of Mortierella sp. (a high-yield fatty acid) according to claim 1 as claimed in claim 3. Mortierella sp. ) The culture medium of 2-1-4 or its application in the preparation of functional foods or medicines.

9. The method for fermenting and culturing a novel high-yield fatty acid-producing bacterium Mortierella as claimed in claim 1 according to claim 4. Mortierella sp. ) Application of the culture medium of 2-1-4 in the preparation of functional foods or medicines.

10. A method for preparing a fatty acid, characterized in that: Using the novel high-fat-yielding fungus Mortierella as claimed in claim 1 ( Mortierella sp. )2-1-4 prepared by fermentation.

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