Phaffia rhodozyma capable of producing astaxanthin at high yield and application thereof

Abstract

The invention relates to the technical field of edible fungus strain breeding, in particular to phaffia rhodozyma capable of producing astaxanthin at high yield and application of the phaffia rhodozyma. The phaffia rhodozyma HFF5011 provided by the invention is preserved in the China Center for Type Culture Collection on December 16, 2024, and the preservation number of the phaffia rhodozyma HFF5011 is CCTCC (China Center for Type Culture Collection) NO: M 20242822. The strain is obtained by domestication and breeding on the basis of phaffia rhodozyma (named as phaffia rhodozyma HFF501) with the preservation number of CCTCC (China Center For Type Culture Collection) NO: M 2020413. The phaffia rhodozyma provided by the invention is a strain with higher astaxanthin yield screened by utilizing a beta-violone culture medium on the basis of an original strain, and the content of astaxanthin fermented by the strain can be up to 630mg / L.

Description

Technical Field

[0001] This application relates to the technical field of strain breeding, and particularly relates to a Phaffia rhodozyma with high astaxanthin production and its application. Background Art

[0002] Phaffia rhodozyma (also known as Phaffia rhodozyma) is a microorganism of the genus Phaffia, native to China, and is mainly used to produce astaxanthin. Astaxanthin is not only a kind of pigment, but also one of the most powerful lipophilic antioxidants discovered so far. It has the unique ability to quench free radicals and reactive species of oxygen, and inhibit lipid peroxidation. A lot of evidence shows that astaxanthin has certain effects in the prevention and treatment of various diseases, including cancer, metabolic syndrome, diabetes, kidney function repair, cardiovascular diseases, neurodegenerative diseases, ophthalmic diseases, skin diseases, gastrointestinal and liver diseases, etc.

[0003] At present, the methods for obtaining high-astaxanthin-producing strains include traditional methods such as physical and chemical mutagenesis and protoplast fusion, as well as many transgenic technologies. For example: Chinese invention patent CN 113789322 B selects astaxanthin-producing strains through methods such as ARTP mutagenesis and β-ionone screening, and obtains a Phaffia rhodozyma X17 with high astaxanthin production, that is, an astaxanthin-producing strain. Another example: Chinese invention patent CN 114561304 B effectively promotes the accumulation of astaxanthin in the cells through a suitable glucose feeding process, adding appropriate precursors during the cell fermentation cycle, and then matching the optimized fermentation process, and the astaxanthin yield reaches more than 600 mg / L.

[0004] Although based on the previous studies, the astaxanthin content of Phaffia rhodozyma has been improved, the method for obtaining high-astaxanthin-producing strains is relatively complex. To meet the needs of industrial production, the method needs to be simplified, and the biomass and astaxanthin content of Phaffia rhodozyma need to be improved. Summary of the Invention

[0005] This application provides a Phaffia rhodozyma with high astaxanthin production and its application. The Phaffia rhodozyma provided in this application is a strain with higher astaxanthin production screened by using a β-ionone medium on the basis of the original strain, and the content of astaxanthin fermented by this strain can reach up to 630 mg / L at most.

[0006] In the first aspect, this application provides a Phaffia rhodozyma with high astaxanthin production, and the following technical scheme is adopted:

[0007] A Phaffia rhodozyma with high astaxanthin production. The Phaffia rhodozyma (Phaffia rhodozyma) HFF5011 was deposited at the China Center for Type Culture Collection on December 16, 2024, with the deposit number CCTCC NO: M20242822, and the deposit address is: Inside Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province.

[0008] The above strain was obtained by domestication and breeding on the basis of Phaffia rhodozyma with the deposit number CCTCC NO: M 2020413 (named Phaffia rhodozyma HFF501).

[0009] Optionally, the Phaffia rhodozyma includes a specific nucleotide sequence shown in any one of SEQ ID NO: 1-2.

[0010] Optionally, the primer sequences for amplifying the specific nucleotide sequence shown in SEQ ID NO: 1 are as shown in SEQ ID NO: 3-4.

[0011] Optionally, the primer sequences for amplifying the specific nucleotide sequence shown in SEQ ID NO: 2 are as shown in SEQ ID NO: 5-6.

[0012] In a second aspect, the present application provides a culture. The culture includes the above Phaffia rhodozyma.

[0013] In a third aspect, the present application provides a fermentation broth. The fermentation broth includes the above Phaffia rhodozyma.

[0014] In a fourth aspect, the present application provides a microbial agent. The microbial agent includes the above Phaffia rhodozyma.

[0015] In a fifth aspect, the present application provides a microbial powder. The microbial powder includes the above Phaffia rhodozyma.

[0016] Centrifuge the fermentation broth obtained by shake flask culture of Phaffia rhodozyma, and keep the supernatant to obtain the fermentation supernatant of Phaffia rhodozyma.

[0017] The fermentation broth obtained by culturing Phaffia rhodozyma in a fermenter is the microbial agent of Phaffia rhodozyma.

[0018] Perform spray drying on the fermentation broth obtained by culturing Phaffia rhodozyma in a fermenter to obtain the microbial powder of Phaffia rhodozyma.

[0019] In a sixth aspect, the present application provides the use of the above Phaffia rhodozyma and its culture, fermentation broth, microbial agent or microbial powder in the field of feed additives.

[0020] In summary, the present application includes at least one of the following beneficial technical effects:

[0021] The Phaffia rhodozyma provided in this application is a strain with higher astaxanthin production screened using a β-ionone medium based on the original strain. The content of astaxanthin fermented by this strain can reach up to 630 mg / L at most.

[0022] In addition, the powder product of astaxanthin also has good feedback in the actual application in shrimp farms, and can enhance the stress resistance and feeding efficiency of aquaculture animals such as shrimp larvae. Description of the Drawings

[0023] Figure 1 It is a comparison chart of the dry weight of yeast cells of Phaffia rhodozyma strains with different domestication batches and the original strain in Example 1 of this application (①, ②, and ③ respectively represent the strains selected on the 300 ppm ionone domestication screening medium for 3 batches).

[0024] Figure 2 It is a comparison chart of the astaxanthin yields of Phaffia rhodozyma strains with different domestication batches and the original strain in Example 1 of this application (①, ②, and ③ respectively represent the strains selected on the 300 ppm ionone domestication screening medium for 3 batches).

[0025] Figure 3 It is a comparison chart of the dry weight of cells at different time points during the fermentation process of the finally domesticated strain and the original strain.

[0026] Figure 4 It is a comparison chart of the astaxanthin fermentation amounts at different time points during the fermentation process of the finally domesticated strain and the original strain. Detailed Embodiments

[0027] Before describing the embodiments of this application in detail, it should be understood that the terms used herein are only for the purpose of describing specific embodiments. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this term belongs.

[0028] It should be noted that the terms "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly specifying the quantity of the indicated technical features. Thus, the features defined with "first" and "second" may explicitly or implicitly include one or more of such features. Further, in the description of this application, unless otherwise stated, the meaning of "a plurality" is two or more.

[0029] In the ranges disclosed in this application, the endpoints and any values are not limited to the exact ranges or values. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, the endpoint values of each range, between the endpoint values of each range and individual point values, and between individual point values can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0030] In this application, the term "comprising" or "including" is an open expression, that is, it includes the content specified in this application, but does not exclude other aspects of the content.

[0031] To make the purpose, technical solutions and advantages of this application clearer, the technical solutions in the embodiments of this application will be described clearly and completely below. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative efforts fall within the scope of protection of this application. The embodiments described below are exemplary and are only used to explain this application, and should not be construed as a limitation of this application.

[0032] For those not specifying specific techniques or conditions in the embodiments, follow the techniques or conditions described in the literature in this field or according to the product specifications. For reagents or instruments not indicating the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0033] The test materials used in this application are all ordinary commercially available products and can be purchased in the market.

[0034] Among them, the method for measuring astaxanthin in yeast cells is specifically as follows:

[0035] (1) Take the sample of the strain fermentation broth and stir evenly. Use a pipette to suck 1 mL of the bacterial liquid into a 5 mL centrifuge tube, centrifuge at 8000 rpm for 5 min, and discard the supernatant; add 2 mL of dimethyl sulfoxide preheated to 60 °C to the centrifuged cells, mix well by pipetting, place in a water bath at 50 °C for cell wall breaking treatment for 5 min, then transfer the broken bacterial liquid to a 15 mL centrifuge tube, add 7 mL of absolute ethanol, and let it stand in the dark for extraction for 10 min;

[0036] (2) Centrifuge the above cell wall broken and extracted pigment cell mixture at 8000 rpm for 5 min, transfer the supernatant to a 50 mL brown volumetric flask, repeat the above extraction steps until the cells are close to white, and all the subsequent collected supernatants are combined into the supernatant of the first extraction, and make up to 50 mL with absolute ethanol.

[0037] (3) After filtering the above extraction solution through a 0.22 μm organic filter membrane, prepare for liquid phase injection.

[0038] Among them, the preparation of astaxanthin mother liquor:

[0039] (1) Weigh 5.00 mg of astaxanthin standard precisely with a Mettler ME55 one in a hundred thousand balance. After dissolving it with dimethylformamide, make up the volume to 100 mL in a brown volumetric flask to obtain a 50 mg / L astaxanthin standard mother liquor, and place it in a refrigerator at 4 °C for standby.

[0040] (2) Dilute the mother liquor with chromatographic grade methanol (Rainin pipette guns, specifications 1000 μL, 200 μL, 100 μL) to prepare astaxanthin standard solutions with concentrations of 0, 0.25 mg / L, 0.5 mg / L, 1 mg / L, 2 mg / L, 4 mg / L, 8 mg / L, and 16 mg / L respectively. After filtering through a 0.22 μm organic filter membrane, prepare for liquid phase injection.

[0041] Among them, the detection of astaxanthin concentration:

[0042] High performance liquid chromatograph: Agilent 1260 Infinity Ⅱ, and the chromatographic conditions are as follows:

[0043] Mobile phase: chromatographic methanol, flow rate 1 mL / min;

[0044] Chromatographic column: EC-C18; injection volume: 10 μL; column temperature: 30 °C; wavelength 478 nm.

[0045] Among them, the method for determining the dry cell weight:

[0046] First, weigh the empty weight of a 5 ml centrifuge tube used for loading the liquid with an analytical balance and record it. Then, pipette 4 ml of the well-shaken fermentation broth into the above centrifuge tube. After centrifuging at 12000 rpm for 2 min to remove the supernatant, place it in an oven at 90 °C for 6 h of drying, and then weigh the dry cell weight of Rhodotorula rubra with an analytical balance.

[0047] The present application will be further described in detail below in combination with the examples and the test results.

[0048] Examples

[0049] Example 1

[0050] This example provides the screening process of Rhodotorula rubra HFF5011.

[0051] Specifically, it includes the following steps:

[0052] (1) Resuscitate Phaffia rhodozyma HFF501 (preservation number: CCTCC NO: M2020413) on YPD solid medium (formulation: 10 g / L yeast extract, 20 g / L bacteriological peptone, 10 g / L glucose, 15 g / L agar, medium pH 6.0). Pick a single colony grown on the above plate and inoculate it into a bacterial YPD shake flask (formulation: 10 g / L yeast extract, 20 g / L bacteriological peptone, 10 g / L glucose, medium pH 6.0). After culturing for 12 h, obtain a bacterial solution;

[0053] (2) Gradient dilute the bacterial solution by a factor of ten to 10 3 、10 4 times. Take 50 μl of the dilution solutions of two gradients respectively and evenly coat them on YPD solid medium containing different concentrations of β-ionone (300 ppm, 500 ppm, 1000 ppm) (formulation: 10 g / L yeast extract, 20 g / L bacteriological peptone, 10 g / L glucose, 15 g / L agar, medium pH 6.0; β-ionone is added to the medium after sterilization). Place it in a biochemical incubator at 22 °C for acclimation culture. Observe the appearance of single colonies after culturing for 5 - 6 d.

[0054] (3) Pick 3 - 10 single colonies grown on the 300 ppm β-ionone plate, and then perform purification, culture, and fermentation respectively. At the same time, measure the dry weight of yeast cells (as shown in Figure 1 ) and the astaxanthin yield (as shown in Figure 2 ) of the original strain (preservation number: CCTCC NO: M 2020413) and the strains screened on the β-ionone plate at different fermentation time periods.

[0055] (4) After 2 - 3 rounds of acclimation, select the yeast strain with the highest astaxanthin fermentation amount as the optimal astaxanthin-producing strain, and name it Phaffia rhodozyma HFF5011. Preserve and store it.

[0056] Example 2

[0057] This example provides the sequencing results of Phaffia rhodozyma HFF5011 obtained after acclimation in Example 1.

[0058] After sequencing, Phaffia rhodozyma HFF5011 obtained after acclimation in Example 1 includes two specific sequences, namely SF161 and SF172.

[0059] Among them, SF161 is the nucleotide sequence shown in SEQ ID NO: 1. The primer sequences for amplifying the specific nucleotide sequence shown in SEQ ID NO: 1 are shown in SEQ ID NOs: 3-4, and the amplified fragment is 297 bp.

[0060] Among them, SF172 is the nucleotide sequence shown in SEQ ID NO: 2. The primer sequences for amplifying the specific nucleotide sequence shown in SEQ ID NO: 2 are shown in SEQ ID NOs: 5-6, and the amplified fragment is 351 bp.

[0061] The above-mentioned strain was deposited with the China Center for Type Culture Collection on December 16, 2024, with the deposit number CCTCC NO: M 20242822, and the deposit address is: No. 299, Bayi Road, Wuchang District, Wuhan City, Hubei Province, on the campus of Wuhan University.

[0062] Example 3

[0063] This example provides a fermentation broth of Phaffia rhodozyma HFF5011.

[0064] The preparation method of the above-mentioned fermentation broth is as follows:

[0065] (1) Activate the domesticated strain HFF5011 (Phaffia rhodozyma HFF5011) in Example 1. Take an appropriate amount of the bacterial solution, streak it on the slant medium, and culture it at 22 °C for 5-7 d.

[0066] (2) Primary seed: Take a loopful of the bacterial colony on the plate and inoculate it into a 250 mL Erlenmeyer flask containing 50 mL of seed medium (bacterial YPD shake flask medium), and culture it on a shaker at 22 °C and 200 rpm for 24 h. This is the primary seed.

[0067] (3) Secondary seed: Take 3 mL of the primary seed liquid and inoculate it into a 250 mL Erlenmeyer flask containing 50 mL of seed medium (bacterial YPD shake flask medium) (inoculation amount 6%), and culture it on a shaker at 22 °C and 200 rpm for 24 h. This is the secondary seed.

[0068] (4) Fermentation shake flask culture: Load 50 mL of medium (bacterial YPD shake flask medium) into a 250 mL Erlenmeyer flask, inoculate the secondary seed liquid at an inoculation amount of 10%, and culture it on a shaker at 22 °C and 200 rpm to obtain the fermentation broth of Phaffia rhodozyma HFF5011.

[0069] Example 4

[0070] This example provides a fermentation supernatant of Phaffia rhodozyma HFF5011.

[0071] The preparation method of the above fermentation supernatant is as follows:

[0072] Ferment the Rhodotorula rubra HFF5011 according to the fermentation method of Example 3 to obtain a fermentation broth. Centrifuge the fermentation broth and retain the supernatant, which is the fermentation supernatant of Rhodotorula rubra HFF5011.

[0073] Example 5

[0074] This example provides a bacterial agent of Rhodotorula rubra HFF5011.

[0075] The preparation process of the above bacterial agent is as follows:

[0076] (1) Activate the domesticated strain HFF5011 (Rhodotorula rubra HFF5011) in Example 1. Take an appropriate amount of the bacterial solution, streak it on a slant medium, and culture it at 22 °C for 5 - 7 d.

[0077] (2) Primary seed: Pick a loopful of the bacterial colony on the plate and inoculate it into a 250 mL Erlenmeyer flask containing 50 mL of seed medium (bacterial YPD shake flask medium). Culture it on a shaker at 22 °C and 200 rpm for 24 h. This is the primary seed.

[0078] (3) Secondary seed: Take 3 mL of the primary seed solution and inoculate it into a 250 mL Erlenmeyer flask containing 50 mL of seed medium (bacterial YPD shake flask medium) (inoculation amount 6%). Culture it on a shaker at 22 °C and 200 rpm for 24 h. This is the secondary seed.

[0079] (4) Fermentation shake flask culture: Place 50 mL of the medium (bacterial YPD shake flask medium) in a 250 mL Erlenmeyer flask, and inoculate the secondary seed solution at an inoculation amount of 10%. Culture it on a shaker at 22 °C and 200 rpm.

[0080] (5) Fermentation tank culture: Inoculate the secondary seed solution into the fermentation tank at an inoculation amount of 10%. Use sodium hydroxide solution or citric acid solution to control the pH at 6.0 - 6.4, the culture temperature is 22 °C, the rotation speed is 100 rpm, the tank pressure is 0.05 ± 0.01 MPa, the air flow rate is 1 vvm, and the fermentation culture time is 168 h to obtain the bacterial agent of Rhodotorula rubra HFF5011.

[0081] Among them, fed-batch fermentation is carried out at a rate of 25 mL / h / L, and the fed-batch medium is glucose 200 - 300 g / L. The fermentation medium is: yeast extract 3 g / L, (NH4)2SO4 0.4 g / L, glucose 15 g / L, sucrose 3 g / L, sodium citrate 2 g / L.

[0082] Example 6

[0083] This embodiment provides a bacterial powder of Rhodotorula rubra HFF5011.

[0084] The preparation process of the above-mentioned bacterial powder is as follows: The bacterial agent obtained in Example 5 is spray-dried to become a bacterial powder, and thus the bacterial powder of Rhodotorula rubra HFF5011 is obtained.

[0085] Performance Detection Test 1

[0086] The domesticated strain HFF5011 (Rhodotorula rubra HFF5011) and the initial strain HFF501 (Rhodotorula rubra HFF501) are respectively fermented and cultured by the fermentation method described in Example 5. Samples are taken on the third day of fermentation for determination of dry cell weight and astaxanthin, and then samples are taken every 24 hours for content detection.

[0087] The test results are as Figure 3 and Figure 4 shown.

[0088] Figure 3 is a comparison chart of the dry cell weight of the final domesticated strain and the original strain at different time points during fermentation. The figure shows that the change in the dry cell weight of the domesticated strain during fermentation is not significant compared with that of the original strain.

[0089] Figure 4 is a comparison chart of the astaxanthin fermentation amounts of the final domesticated strain and the original strain at different time points during fermentation. The results show that, under the same other fermentation conditions, the domesticated strain HFF5011 takes less time to reach the astaxanthin fermentation amount of the original strain HFF501.

[0090] In summary, the domesticated strain (Rhodotorula rubra HFF5011) obtained by domestication in this application takes less time to ferment the same content of astaxanthin compared with the original strain (Rhodotorula rubra HFF501), and can significantly increase the astaxanthin fermentation content, and the astaxanthin content of Rhodotorula rubra HFF5011 can reach up to 630 mg / L at most.

[0091] Example 7

[0092] The bacterial agents of the domesticated strain HFF5011 (Rhodotorula rubra HFF5011) and the initial strain HFF501 (Rhodotorula rubra HFF501) are respectively spray-dried to become bacterial powders by the method described in Example 6.

[0093] Among them, the bacterial content in the bacterial powder corresponding to the domesticated strain HFF5011 (Rhodotorula rubra HFF5011) is 3.2×10 10 cfu / g; the bacterial content in the bacterial powder corresponding to the initial strain HFF501 (Rhodotorula rubra HFF501) is 3.2×10 10cfu / g.

[0094] Performance Detection Test Two

[0095] Choose to use shrimp ponds to compare the effects of domesticated strain HFF5011 bacterial powder and the initial strain HFF501 bacterial powder.

[0096] The address of the shrimp pond is: Guangdong Shrimp Seedling Farm. Select shrimp seedlings of 0.5 - 1 cm to conduct the ability test of astaxanthin extraction resistance.

[0097] The specific test method is as follows: First, take out the shrimp seedlings 20 minutes before they eat and place them in 7 test barrels, with about 1000 shrimp seedlings in each barrel. Subsequently, when feeding the bait, experimental groups 1 and 2 were respectively added with the initial strain bacterial powder with a final concentration of 0.4% and the domesticated bacterial agent bacterial powder with a final concentration of 0.4%; experimental groups 3 and 4 were respectively added with the initial strain bacterial powder with a final concentration of 0.7% and the domesticated bacterial agent bacterial powder with a final concentration of 0.7%; experimental groups 5 and 6 were respectively added with the initial strain bacterial powder with a final concentration of 1.0% and the domesticated bacterial agent bacterial powder with a final concentration of 1.0%; experimental group 7 was the blank control group, and then let it stand for 30 minutes.

[0098] (1) Test for resistance to fresh - water stress: Take the shrimp seedlings of each experimental group and the blank control group, and put them into test barrels filled with 1 L of fresh water respectively. Two parallels were set for each experimental group and the blank control group, and there were 50 shrimp seedlings in each of the two parallels of each group. Record the number of dead shrimp seedlings in each group every 5 minutes for 20 minutes. The results are shown in Table 1.

[0099] Table 1 Test Results of Resistance to Fresh - water Stress

[0100]

[0101] As can be seen from Table 1, after being treated in fresh water for 20 minutes, the survival rate of shrimp seedlings in the blank control group was 12%. On the premise of the same amount of bacterial powder added, the survival rate of shrimp seedlings with domesticated strain bacterial powder was higher than that of the initial bacterial agent bacterial powder; and the survival rate of shrimp seedlings with 0.4% domesticated strain bacterial powder was equivalent to that of shrimp seedlings with 1% initial bacterial agent bacterial powder. It shows that under the condition of the same amount of bacterial powder added, the effect of domesticated strain bacterial powder is better than that of the initial bacterial agent bacterial powder, and both bacterial agents can improve the resistance of shrimp seedlings to fresh - water stress.

[0102] (2) Test for resistance to disinfectant stress: Take 50 shrimp seedlings from each experimental group and the blank control group, and place them in test water buckets added with chlorine dioxide with a final concentration of 5 ppm respectively and let them stand for 30 minutes. Three parallels were set for each group. Record the survival rate of shrimp seedlings in each group after 30 minutes. The results are shown in Table 3.

[0103] Table 3 Test Results of Resistance to Disinfectant Stress

[0104]

[0105] As can be seen from Table 3, after standing still in water containing 5 ppm chlorine dioxide for 30 min, the average recovery rate of the shrimp fry in the blank control group was only 32%, and the recovery rates of the experimental groups with the addition of two kinds of bacterial powders were both above 35%, indicating that feeding the two kinds of bacterial agents can improve the stress resistance of shrimp fry to disinfectants. In addition, on the premise of the same addition amount of bacterial powder, the recovery rates of the shrimp fry with the domesticated strain bacterial powder were higher than those of the shrimp fry with the initial bacterial agent bacterial powder.

[0106] In summary, the application examples of astaxanthin show that during the cultivation process of shrimp fry, appropriately adding yeast agents containing astaxanthin can significantly improve the stress resistance of shrimp fry under environmental changes and the tolerance to chemical drugs such as disinfectants. And from this, it can be known that during the cultivation process of shrimp fry, appropriately selecting Phaffia rhodozyma with higher astaxanthin production can not only reduce costs, but also be more beneficial to the cultivation process of shrimp fry.

[0107] In the description of this specification, the description with reference to terms such as "one embodiment", "some embodiments", "example", "specific example" or "some examples" means that the specific features, structures, materials or characteristics described in connection with the embodiment or example are included in at least one embodiment or example of the present application. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner. In addition, without contradiction, those skilled in the art can combine and combine the different embodiments or examples described in this specification and the features of different embodiments or examples.

[0108] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, not to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements for some of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A Phaffia rhodozyma with high astaxanthin production, characterized in that: The rhodozyma yeast ( Phaffia rhodozyma ) HFF5011 is deposited in China Center for Type Culture Collection with the deposit number CCTCC NO: M 20242822.

2. The rhodozyma yeast according to claim 1, characterized in that The rhodozyma yeast comprises a specific nucleotide sequence as shown in any one of SEQ ID NOs: 1-2.

3. The rhodozyma yeast according to claim 2, characterized in that The primer sequences used to amplify the specific nucleotide sequence shown in SEQ ID NO: 1 are shown in SEQ ID NO: 3-4.

4. The rhodozyma yeast according to claim 2, characterized in that The primer sequences used to amplify the specific nucleotide sequence shown in SEQ ID NO: 2 are shown in SEQ ID NO: 5-6.

5. A culture, characterized in that The culture comprises the rhodozyma yeast of claim 1.

6. A fermentation broth, characterized in that: The fermentation broth comprises the rhodozyma yeast according to claim 1.

7. A bacterial agent, characterized in that The bacterial agent comprises the rhodozyma yeast according to claim 1.

8. A bacterial powder, characterized in that: The bacterial powder comprises the rhodozyma yeast according to claim 1.

9. Use of the rhodozyma yeast according to claim 1, the culture according to claim 5, the fermentation liquid according to claim 6, the bacterial agent according to claim 7, and the bacterial powder according to claim 8 in the field of feed additives.

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