CN113308387A - Bacterial strain for co-production of unsaturated fatty acid and carotenoid and application thereof - Google Patents
Bacterial strain for co-production of unsaturated fatty acid and carotenoid and application thereof Download PDFInfo
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- CN113308387A CN113308387A CN202110345911.8A CN202110345911A CN113308387A CN 113308387 A CN113308387 A CN 113308387A CN 202110345911 A CN202110345911 A CN 202110345911A CN 113308387 A CN113308387 A CN 113308387A
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- rhodosporidium toruloides
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- carotenoid
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- 235000021466 carotenoid Nutrition 0.000 title claims abstract description 20
- 150000001747 carotenoids Chemical class 0.000 title claims abstract description 20
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 9
- 235000021122 unsaturated fatty acids Nutrition 0.000 title claims abstract description 8
- 150000004670 unsaturated fatty acids Chemical class 0.000 title claims abstract description 8
- 230000001580 bacterial effect Effects 0.000 title claims description 15
- 238000000855 fermentation Methods 0.000 claims abstract description 42
- 230000004151 fermentation Effects 0.000 claims abstract description 42
- 241000221523 Rhodotorula toruloides Species 0.000 claims abstract description 41
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 12
- 239000008103 glucose Substances 0.000 claims abstract description 12
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- IPCSVZSSVZVIGE-UHFFFAOYSA-N hexadecanoic acid Chemical compound CCCCCCCCCCCCCCCC(O)=O IPCSVZSSVZVIGE-UHFFFAOYSA-N 0.000 description 8
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- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 description 4
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- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 description 4
- QPJVMBTYPHYUOC-UHFFFAOYSA-N methyl benzoate Chemical compound COC(=O)C1=CC=CC=C1 QPJVMBTYPHYUOC-UHFFFAOYSA-N 0.000 description 4
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- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
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- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Tropical Medicine & Parasitology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Animal Husbandry (AREA)
- Virology (AREA)
- Food Science & Technology (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a strain for co-producing unsaturated fatty acid and carotenoid and application thereof, wherein the strain is classified and named as rhodosporidium toruloides (rhodosporidium toruloides)Rhodosporidiumtoruloides) Z11, the preservation number is CCTCC NO: m2021226. The mutant strain obtained by ion beam mutagenesis in the invention takes glucose as a unique carbon source, and the oil content and the carotenoid yield in the shake flask fermentation respectively reach 66 percent and 13.5mg/g which are respectively 2.12 times and 12.85 times of the original strain cultured under the same conditions; the product has high stability, high yield and good repeatability, and is an excellent strain suitable for being used as animal feedAnd (5) strain. The microbial oil is rich in a large amount of protein and a certain amount of vitamin carotene, is simple in preparation process as an animal additive, has wide sources of production raw materials, is beneficial to realizing large-scale production, solves the problem that the residual mushroom dregs after microbial oil extraction are not suitable for animal digestion and absorption, reduces the production cost of the microbial oil, and promotes the comprehensive utilization of resources.
Description
Technical Field
The invention belongs to the technical field of biological fermentation, and relates to a bacterial strain for co-producing unsaturated fatty acid and carotenoid and application thereof.
Background
Nowadays, with the development of economy, the quality of human life is greatly improved, healthy diet is advocated, the animal husbandry and the breeding industry are rapidly improved, if the animal husbandry and the breeding industry are fed in a traditional mode, the obtained protein cannot reach a high-quality grade, and therefore the problem of the shortage of protein feed supply is solved by using mycoprotein as a substitute.
Mycoprotein, also known as microbial protein, is usually obtained by culturing unicellular microorganisms, and is a microbial biomass artificially cultured from many industrial and agricultural wastes as well as petroleum wastes. Thus, a single-cell protein is not a pure protein, but a cytoplasmic mass consisting of a mixture of proteins, fats, carbohydrates, nucleic acids, and nitrogen-containing compounds, vitamins, and inorganic compounds that are not proteins. Important among the single-cell proteins are yeast proteins, bacterial proteins and algal proteins, and their chemical composition is mainly protein and fat. The protein content can reach 50-80%, so that the application of the single-cell protein in the feed has high application value.
A major important component in single-cell protein is vitamin, which is a kind of trace organic substance that human and animal must obtain from food in order to maintain normal physiological function, playing an important role in the growth, metabolism and development process of human body. It has the function of regulating the metabolism of organism. But it is different from proteins, lipids and carbohydrates. The human body and the animal body can not be synthesized automatically, and the human body and the animal body are required to be obtained from food, and if the vitamin is deficient, the metabolism, the growth, the development and the health of the body are influenced to a certain extent.
In China, the oleaginous microorganisms include bacteria, yeast, fungi, microalgae and the like, and the microorganisms utilize carbohydrates, hydrocarbons and common oil as carbon sources and nitrogen sources, oil produced by inorganic salts as auxiliary materials and other commercially valuable lipids under certain conditions. Wherein if the intracellular accumulated oil is 20% (w/w) of the total organism, it is called oleaginous microorganism. Wherein in eukaryotic microorganisms, certain oleaginous species can accumulate intracellularly to a fat content of 70% (w/w) of their total biological quantity. The oil exists in cells mainly in the form of fatty acid triglyceride, and the composition of the oil is similar to that of common animal and vegetable oil, so that the microbial oil fermentation has important research significance in the aspect of industrial production of biodiesel and biological feed.
Since the microbial oil is an intracellular product, a large amount of thallus residue is accompanied when the oil is obtained. The residue left after extraction of the oil is used as a potential source of feed protein, whether it is a prokaryotic or eukaryotic microorganism. In particular, yeast, mold, microalgae and the like which produce high amounts of fats and oils have attracted attention in recent years because of their high fat and oil contents and high biosafety. Researches on producing microbial oil by fermenting various non-grain crops, agricultural and forestry wastes, industrial waste liquid, industrial wastes and the like by using the oil-producing microorganisms are increasing.
The extraction methods of microbial oil commonly used in industry include squeezing, leaching and supercritical fluid extraction. The pretreatment for extracting oil is also important, and different pre-treatment methods for producing strains are also commonly adopted, such as drying the strains and then carrying out high-temperature treatment, or carrying out enzymolysis, physical crushing or chemical treatment and the like to crack the bacterial walls. Some of the wall breaking methods are only suitable for extracting a small amount of broken oil in a laboratory, if a large amount of industrial production is carried out, the economic cost is a big factor, and some methods achieve the effect of breaking the bacterial wall, but chemical reagents of the methods remain on the bacteria, so that the recycling value of the bacterial residues is greatly reduced. Therefore, it is very important to extract oil from the bacteria in large quantities and adopt a proper oil extraction method and a proper bacteria residue post-treatment method for different bacteria. The method is not only beneficial to accelerating the industrial production process of industries such as microbial oil and biodiesel, but also can promote the comprehensive utilization of resources and solve the problem of supply shortage of raw materials for producing mycoprotein.
The microbial oil fermentation residues contain a large amount of protein and cell wall components. In general, the residue obtained by squeezing or supercritical fluid extraction of the cells has no solvent residue, and can be used as a cell protein feed. Therefore, the problem of reutilization rate of fungus dregs due to oil extraction is also considered when the process of high yield of carotenoid is achieved, and finally large-scale industrial production is realized.
Disclosure of Invention
The invention aims to provide a bacterial strain for co-producing unsaturated fatty acid and carotenoid and application thereof.
In order to achieve the technical purpose, the technical scheme adopted by the invention is as follows:
a bacterial strain for co-producing unsaturated fatty acid and carotenoid is classified and named as Rhodosporidium toruloides Z11 with the preservation number of CCTCC NO: m2021226.
The invention also aims to provide the application of the rhodosporidium toruloides in producing the grease.
Specifically, the rhodosporidium toruloides is obtained by extracting a fermentation liquid obtained after fermentation culture of the rhodosporidium toruloides by using an organic solvent after freeze drying.
Still another object of the present invention is to provide the use of said Rhodosporidium toruloides for the production of carotenoids.
The invention also aims to provide application of the rhodosporidium toruloides in preparing animal feed additives.
Specifically, after freeze drying of fermentation liquor obtained after fermentation culture of rhodosporidium toruloides, oil is extracted by an organic solvent, and after hydrolysis treatment, a liquid additive finished product with the total solid concentration of 10-90% w/w is prepared or a solid additive finished product is prepared by drying at 20-100 ℃.
The fermentation culture mode of the rhodosporidium toruloides is as follows:
1) plate culture: inoculating rhodosporidium toruloides to a plate culture medium for culture at the culture temperature of 28-30 ℃ for 24-30 h;
2) seed culture: inoculating the rhodosporidium toruloides cultured by a plate into a seed culture medium for culturing at the culture temperature of 28-30 ℃ for 24-30 h;
3) fermentation culture: inoculating the seed culture solution into a fermentation culture medium, wherein the inoculation amount is 10-12% v/v, the fermentation temperature is 28-30 ℃, and the fermentation culture time is 100-120 h.
Further, the rhodosporidium toruloides is fermented and cultured under an aerobic condition.
Further, the rhodosporidium toruloides is fermented and cultured by taking glucose as a carbon source.
Further, the culture medium for the rhodosporidium toruloides fermentation culture comprises the following components in percentage by mass: glucose, K2HPO40.04％、NaH2PO40.1％、MgSO4·H20.15% of O and 0.175% of yeast extract, wherein the trace elements are: ZnSO4、CaCl2、MnCl2、CuSO4Are respectively 0.192 × 10-7g/L、0.167g/L、0.54×10-4g/L、0.16×10-4g/L。
The invention adopts ion beams to mutate rhodosporidium toruloides, and utilizes a flat plate to screen strains which have stronger tolerance and can produce high carotenoid yield. The strain and the process are utilized for fermentation, glucose is used as a carbon source, the oil content and the carotenoid yield in a shake flask fermentation product respectively reach 66% and 13.5mg/g which are respectively 1.57 times and 12.85 times of the original strain cultured under the same conditions, and the oil component analysis shows that the linoleic acid and the palmitic acid account for 68% and 22% of the total oil ratio respectively, so that the strain and the process can be used for co-producing the linoleic acid and the palmitic acid; has great social significance and economic value.
Detailed Description
The invention will be better understood from the following examples. However, it is easily understood by those skilled in the art that the embodiments are described only for illustrating the present invention and should not limit the present invention described in detail in the claims.
The biological material is classified and named as Rhodosporidium toruloides Z11, is preserved in China center for type culture Collection CCTCC, and has a preservation number of CCTCC NO: m2021226, preservation date of 2021, 03 and 15 days, preservation address: wuhan in China.
Example 1: this example illustrates a method for screening Rhodosporidium toruloides.
The Rhodosporidium toruloides original strain is wild strain screened from soil collected in lake near Aster canteen of Nanjing university of industry;
specifically, YPD plates are used as culture media, the culture is carried out under the aerobic condition at 30 ℃, the culture is carried out for 2-4 days, streaking and purifying are carried out for 5-7 times, finally, strains which can grow on the plates are verified, the strains are fermented, fermentation products and performances of the strains are examined, and the strains can grow by utilizing a plurality of carbon sources and can produce a trace amount of carotenoid. The strain is used as an original strain to carry out strain transformation.
The first ion beam mutagenesis was performed as follows:
activating and culturing an original strain of rhodosporidium toruloides at the culture temperature of 30 ℃, the liquid loading amount of a 100mL shake flask is 20mL, and the culture time is 24-30 h to obtain a bacterial liquid in a logarithmic phase; freshly cultured cells were diluted with physiological saline to OD cell concentration600Spreading 10 microliter of the bacterial liquid on a small iron sheet, and drying by sterile air; implant 1.6X 10 with 10KeV16ions/cm2Dose, pulse mode implant 5s each, interval 15 s. After ion implantation mutagenesis, the biofilm was eluted.
The mutagenized strains were further screened as follows:
wherein the formula of the used culture medium (% by mass):
(1) solid plate medium: 1% of yeast powder, 2% of peptone, 2% of glucose, 2% of agar and the balance of water, wherein the pH value is 6.
(2) Shake flask fermentation screening culture medium: glucose 8%, K2HPO4 0.04％、NaH2PO4 0.1％、MgSO4·H20.15% of O, 0.075% of yeast extract, adding different carbon sources (peptone, glutamic acid, glacial acetic acid, ammonium sulfate), pH 6.0, 115 deg.C, and autoclaving for 20 min. Note: removing acetic acid and MgSO4·H2And O is the filtration sterilization.
(3) Phosphate buffer solution: 0.2M NaH2PO4The solution was autoclaved at 121 ℃ for 20min until the pH was 6.0.
A screening step:
1. plate screening
And (3) washing out the strains subjected to ion beam mutagenesis by using physiological saline, diluting the strains into different concentrations, coating the different concentrations on a conventional YPD solid medium plate, carrying out aerobic culture at the temperature of 28-30 ℃ for 30h, and selecting a plurality of colonies which can grow on a screening plate and have larger colonies.
2. Flat-plate double screen
Inoculating the screened strain into a 100mL triangular flask with the liquid loading amount of 20mL, carrying out aerobic culture at the temperature of 28-30 ℃ for 24-30 h, and preparing sterile physiological saline into OD with concentration600Sucking 2uL of the bacterial suspension liquid on a spot culture medium plate, carrying out aerobic culture at the temperature of 28-30 ℃ for 12-24 h, and selecting out bacterial colonies with the bacterial colonies larger than those of the grown-out bacteria.
The final strains Z11 and Z12 produced the greatest amount of carotenoids.
3. Shake flask fermentation screening
Inoculating the strains Z11, Z12 and the original strain into a seed culture medium for amplification culture, wherein the culture temperature is 30 ℃, the liquid loading capacity of a 250mL triangular flask is 50mL, and the culture time is 24 h. Then, the fermentation medium is used for aerobic fermentation, the inoculation amount is 10 percent (v/v), the fermentation temperature is 30 ℃, the liquid loading amount of a 500mL triangular flask is 100mL, and the carotenoid yield of each strain is detected after the fermentation time is 120h and is shown in Table 1:
TABLE 1 screening of the results of fermentation of Z11, Z12 and the original bacteria in the culture Medium
The carotenoid yield and the oil content of the two mutant strains obtained by plate combination screening are obviously higher than those of the original strain in the fermentation process, wherein Z11 has the highest yield. This is consistent with the results of the combinatorial plate screen.
Example 2: this example illustrates the passaging stability of mutants Z11 and Z12.
The passage stability of mutants Z11 and Z12 was tested in a fermentation medium with glucose as carbon source. The results of the passaged fermentation tests of strains Z11 and Z12 are shown in table 2:
TABLE 2 results of the fermentation test for strains Z11 and Z12 at passages
From the experimental results, the carotenoid yield of the two mutant strains is stable after 7 times of continuous passage, the two mutant strains have good passage stability, and the two mutant strains can be used as production strains for further research and development.
Example 3: this example illustrates a process for the fermentative production of carotenoids by Rhodosporidium toruloides Z11.
The formulation of the culture medium described in this example (% by mass):
plate culture medium: 1% of yeast powder, 2% of peptone, 2% of glucose, 2% of agar and the balance of water, wherein the pH value is 6.
Seed culture medium: 1% of yeast powder, 2% of peptone, 2% of glucose and the balance of water.
Fermentation medium: glucose 8%, K2HPO4 0.04％、NaH2PO40.1％、MgSO4·H20.15% of O and 0.175% of yeast extract, wherein the trace elements are: ZnSO4、CaCl2、MnCl2、CuSO4Are respectively 0.192 × 10-7g/L、0.167g/L、0.54×10-4g/L、0.16×10-4g/L。
Rhodosporidium toruloides Z11 was inoculated into a plate medium and cultured aerobically at 30 ℃ for 26 hours. Inoculating the Z11 subjected to plate culture into a seed culture medium, wherein the liquid loading capacity of a 250mL triangular flask is 50mL, the culture temperature is 30 ℃, and the culture time is 26 h; inoculating the seeds into a fermentation culture medium, wherein the inoculation amount is 10% (v/v), the fermentation temperature is 28-30 ℃, the liquid loading amount of a 500mL triangular flask is 100mL, and after aerobic fermentation culture is carried out for 120h, the detected carotenoid reaches 13.5mg/g, which is 12.85 times higher than that of the original strain cultured under the same conditions.
Example 4: this example shows that Rhodosporidium toruloides Z11 has a stable and high-yielding oil content in part.
Collecting the fermented bacteria liquid (Z11, Z12) completely, freeze-drying for 48H, weighing 20mg of strain lyophilized powder in a 15mL glass tube, adding 1.5mL of n-hexane, 0.5mL of methyl benzoate and 2mg/mL of methyl benzoate, dissolving in n-hexane, and adding 2mL of 15% H2SO4(esterification) in methanol.
The pretreated sample was placed in an environment at 100 ℃ for 2h with continuous shaking. Then placed on ice to cool for 10min, and 1mL ddH was added2And O, centrifuging at 2500rpm for 5 min. The supernatant (fatty acid methyl ester extract) was prepared, placed in a GC flask, and subjected to gas phase detection. The total oil content and the contents of palmitic acid and linoleic acid of each strain are detected as shown in the following table:
TABLE 3 results of the fermentation test for strains Z11 and Z12 at passages
The experimental results show that the total oil content of the strain Rhodosporidium toruloides Z11 reaches 66%, which is 2.12 times higher than that of the original strain under the same culture conditions, wherein the palmitic acid and linoleic acid in the oil content account for 22% and 68% of the total oil ratio, and are 20-30 times higher than that of similar unsaturated fatty acids produced by other Rhodosporidium toruloides.
Example 5: this example illustrates the production of solid mushroom dregs from Rhodosporidium toruloides Z11 for use as animal feed.
In the method of example 4, Rhodosporidium toruloides Z11 was cultured by fermentation, and the fermentation broth was freeze-dried to obtain 58.5g of dry cells having an oil content of 66%. Grinding dry thallus into powder in a grinder, adding 0.1L n-hexane into 50g of thallus powder for extraction, ultrasonically oscillating for 5-15min, centrifuging at 6000rpm for 10min to obtain upper layer mixed oil, cooking the mixed oil to obtain 39g of yeast oil, performing desolventizing treatment, and recovering solid mushroom residue. Mixing the mushroom dregs with a proper amount of water to adjust the total solid concentration to 90%, adjusting the pH value to be within the range of 3.0-4.5 by using sulfuric acid, continuously shaking for 20 minutes at the temperature of 90 ℃, cooling to the room temperature on ice, and neutralizing to the pH value of 5.5-6.0 by using a 4M NaOH solution. And continuously adding water to adjust the concentration of the total solid matters to be 30 percent to obtain 130mL of finished product of the liquid animal feed additive. The product is brown suspension with slight fragrance, and contains crude fat 5.2%, crude protein 12%, crude fiber 4%, and ash 2.3%.
Claims (10)
1. A bacterial strain for coproducing unsaturated fatty acid and carotenoid is characterized in that the bacterial strain is named as rhodosporidium toruloides (A)Rhodosporidiumtoruloides) Z11, the preservation number is CCTCC NO: m2021226.
2. Use of Rhodosporidium toruloides according to claim 1 for the production of fats and oils.
3. The use of claim 2, wherein the fermented liquid obtained by fermenting and culturing the rhodosporidium toruloides is obtained by freeze-drying and extracting with an organic solvent.
4. Use of Rhodosporidium toruloides according to claim 1 for the production of carotenoids.
5. Use of Rhodosporidium toruloides according to claim 1 for the preparation of an animal feed additive.
6. The use of claim 5, wherein the liquid fermented product of Rhodosporidium toruloides fermentation culture is freeze-dried, extracted with an organic solvent to obtain fat, and hydrolyzed to produce a liquid additive product having a total solid-content of 10-90% w/w or a solid additive product dried at 20-100 ℃.
7. The use according to any one of claims 2 to 6, wherein the Rhodosporidium toruloides is cultured by fermentation in the following manner:
1) plate culture: inoculating rhodosporidium toruloides to a plate culture medium for culture at the culture temperature of 28-30 ℃ for 24-30 h;
2) seed culture: inoculating the rhodosporidium toruloides cultured by the plate into a seed culture medium for culturing, wherein the culture temperature is 28-30 ℃, and the culture time is 24-30 h;
3) fermentation culture: inoculating the seed culture solution into a fermentation culture medium, wherein the inoculation amount is 10-12% v/v, the fermentation temperature is 28-30 ℃, and the fermentation culture time is 100-120 h.
8. The use according to any one of claims 2 to 6, wherein the Rhodosporidium toruloides is cultured by fermentation under aerobic conditions.
9. The use of any one of claims 2 to 6, wherein the Rhodosporidium toruloides is cultured by fermentation using glucose as a carbon source.
10. The use of any one of claims 2 to 6, wherein the culture medium for the rhodosporidium toruloides fermentation culture comprises the following components in percentage by mass: glucose, K2HPO4 0.04%、NaH2PO40.1%、MgSO4·H20.15% of O and 0.175% of yeast extract, wherein the trace elements are:ZnSO4、CaCl2、MnCl2、CuSO4are respectively 0.192 × 10-7g/L、0.167g/L、0.54×10-4g/L、0.16×10-4g/L。
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