CN107937280B - Schizochytrium limacinum and application thereof - Google Patents

Schizochytrium limacinum and application thereof Download PDF

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CN107937280B
CN107937280B CN201711213889.1A CN201711213889A CN107937280B CN 107937280 B CN107937280 B CN 107937280B CN 201711213889 A CN201711213889 A CN 201711213889A CN 107937280 B CN107937280 B CN 107937280B
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schizochytrium
schizochytrium limacinum
fermentation
dha
astaxanthin
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CN107937280A (en
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吴永刚
曾曾
孔祥瑞
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Sichuan Chaosheng Biotechnology Co ltd
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    • 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
    • C12N1/145—Fungal isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • 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
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • 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
    • C12P7/6409—Fatty acids
    • C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone

Abstract

The invention relates to the field of microbial fermentation engineering, in particular to schizochytrium limacinum and application thereof; the schizochytrium limacinum is classified and named as schizochytrium sp, the laboratory name is schizochytrium limacinum SC01 strain, the schizochytrium limacinum has been preserved in China general microbiological culture Collection center (CGMCC), the preservation number is CGMCC No.14574, and the preservation date is 2017, 8 and 29 days; the schizochytrium limacinum strain is obtained by separating kelp residues collected from Shandong Ronghai by a pine pollen fishing method, is rich in DHA and astaxanthin, can be directly used as food, health-care products or food additives, and can also be used as feed additives for aquatic products, livestock breeding and the like.

Description

Schizochytrium limacinum and application thereof
Technical Field
The invention relates to the field of microbial fermentation engineering, in particular to schizochytrium limacinum and application thereof.
Background
Schizochytrium sp is a marine fungus belonging to the family Thraustochytriaceae, and has the characteristics of rapid growth, strong stress resistance, high lipid content and the like. Widely distributed in organic substances in seaports, beaches and mangrove areas. Lipid products with high added value, such as docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), astaxanthin and the like, which have high nutritional value, are contained in intracellular oil, and the production of DHA by using schizochytrium of different strains has been realized in Europe, China, Japan and the like.
Docosahexaenoic Acid (DHA) is a polyunsaturated fatty Acid essential to the human body and must be supplemented directly or indirectly from the diet. DHA is an important structural lipid component of central nervous system and retina, has effects of nourishing brain, improving memory and eyesight, especially promoting brain cell development and growth of fetus and infant, improving memory of teenager, and preventing and treating senile dementia.
Astaxanthin (Astaxanthin), a carotenoid, is a powerful natural antioxidant. Astaxanthin, like other carotenoids, is a lipid-soluble and water-soluble pigment. The astaxanthin has strong oxidation resistance which is 550 times that of vitamin E and 10 times that of beta-carotene. Has effects in protecting skin and eyes, resisting radiation, and preventing cardiovascular aging, senile dementia and cancer.
In view of the important effects of docosahexaenoic acid and astaxanthin on human development, cardiovascular and the like, the appropriate supplement of adults, particularly middle-aged and elderly people, is of great benefit. Functional food of non-animal origin containing DHA and astaxanthin will have a wide market. The schizochytrium can synthesize DHA and astaxanthin when heterotrophically cultured in the presence of continuous fluorescence. The prior patent relating to the screening of schizochytrium limacinum strains and the preparation of DHA and astaxanthin by fermentation mainly relates to four aspects: (1) breeding strains for producing DHA excellent strains, such as patent CN 102899254A, CN 102888348A; (2) optimizing the formula of the culture medium, such as patents: CN 101831387B, CN 101886044A; (3) the production cost is reduced and the product yield is improved by optimizing a fermentation process mode, such as patent CN 101812484B, CN 103614427A; (4) through special illumination and nutrition conditions, the schizochytrium produces DHA and astaxanthin at the same time, for example, patent CN 1276073C, CN 104185678A. However, in view of the above patents, the culture conditions for producing DHA and astaxanthin simultaneously by schizochytrium limacinum are harsh, and require special illumination conditions, and providing sufficient illumination for cells under high-density heterotrophic fermentation conditions is difficult to achieve in industrial production, and the astaxanthin content is low, about 0.1% -0.2%, which results in high production cost and serious hindrance to application and popularization. Therefore, the selection of an industrial production strain capable of simultaneously accumulating DHA and astaxanthin under mild conditions, particularly under conditions that do not require light, is a key to industrial fermentation production.
Disclosure of Invention
The invention aims to provide a schizochytrium strain capable of simultaneously producing DHA and astaxanthin without illumination conditions and a method for producing functional oil with high added value by fermenting the strain. The invention screens and obtains a schizochytrium sp SC01(CGMCC No.14574) strain capable of simultaneously producing DHA and astaxanthin from Shandong Rongcheng sea area, and has good industrial application value.
The technical scheme of the invention is as follows:
the schizochytrium limacinum is classified and named as schizochytrium sp, and the laboratory is named as schizochytrium limacinum SC01 strain, and has been preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.14574 and the preservation date of 2017, 8 and 29 days.
The Schizochytrium limacinum strain is obtained by separating the kelp residue collected from Shandong Rongcheng sea area by pollen Pini fishing method. Observed by an optical microscope, the following results are found: the thallus is spherical or ellipsoidal, has a diameter of 8-22 microns, is orange red, and is darkened along with the prolonging of the culture time, and has double flagella with different lateral lengths, and intracellular fatty acids mainly comprise hexadecanoic acid, octadecanoic acid, arachidonic acid, docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA), wherein the DHA content accounts for more than 50% of the total fatty acids, and the astaxanthin content is about 0.3%. Comparing the 18S rDNA sequence by a molecular biology means, finding that the homology of the strain and the 18S rDNA sequence of the schizochytrium is more than 99 percent, thereby judging that the strain belongs to the schizochytrium sp.SC01 and being named as Aurantiochytrium sp.SC01 in a laboratory.
Use of Schizochytrium limacinum for the preparation of docosahexaenoic acid (DHA). Specifically, the DHA grease is prepared by taking the schizochytrium limacinum as an initial strain and performing high-density fermentation in a fermentation medium.
The fermentation culture comprises the following steps:
a. inoculating the strain stored in the glycerinum pipe into a 250ml shake flask filled with 50ml of seed culture medium, and culturing for 24h in a shaking table at the rotating speed of 200rpm to obtain first-grade seeds;
b. inoculating the primary seeds into a 500ml shake flask filled with 100ml of seed culture medium, and culturing for 12h in a shaking table at the rotating speed of 200rpm to obtain secondary seeds;
c. adding a fermentation culture medium into the bioreactor, and inoculating the activated secondary seed liquid.
In the fermentation process, the temperature is controlled to be 20-30 ℃, in the fermentation process, nitrogen supplement operation is adopted, and 20% (w/v) of yeast powder solution or corn steep liquor solution is fed in 36-64 hours of fermentation. The pH was maintained at 6.5 by automatic addition of 2M NaOH. The fermentation time is 80-100 hours.
Wherein: the seed culture medium contains 10-40g/L glucose, 5-15g/L yeast extract and 5-30g/L seawater crystal;
the fermentation culture medium contains 20-80g/L glucose, 5-35g/L yeast extract, 1-5g/L corn steep liquor, 0.3-4g/L potassium dihydrogen phosphate, 0.2-3g/L magnesium sulfate, 5-30g/L seawater crystal, 5-20 mg/L vitamin B13, 20-20 mg/L vitamin B63, 10-10 mg/L vitamin B121 and 1-10mg/L biotin.
In the fermentation process, sugar supplement is adopted, the glucose concentration in the system is controlled to be between 5 and 10g/L, and the glucose concentration is not higher than 3g/L after the fermentation is finished.
An application of Schizochytrium limacinum in preparing astaxanthin is provided. Specifically, the astaxanthin is prepared by taking the schizochytrium limacinum as an initial strain and performing high-density fermentation in a fermentation medium.
The fermentation culture comprises the following steps:
a. inoculating the strain stored in the glycerinum pipe into a 250ml shake flask filled with 50ml of seed culture medium, and culturing for 24h in a shaking table at the rotating speed of 200rpm to obtain first-grade seeds;
b. inoculating the primary seeds into a 500ml shake flask filled with 100ml of seed culture medium, and culturing for 12h in a shaking table at the rotating speed of 200rpm to obtain secondary seeds;
c. adding a fermentation culture medium into the bioreactor, and inoculating the activated secondary seed liquid.
In the fermentation process, the temperature is controlled to be 20-30 ℃, in the fermentation process, nitrogen supplement operation is adopted, and 20% (w/v) of yeast powder solution or corn steep liquor solution is fed in 36-64 hours of fermentation. The pH was maintained at 6.5 by automatic addition of 2M NaOH. The fermentation time is 80-100 hours.
Wherein: the seed culture medium contains 10-40g/L glucose, 5-15g/L yeast extract and 5-30g/L seawater crystal;
the fermentation culture medium contains 20-80g/L glucose, 5-35g/L yeast extract, 1-5g/L corn steep liquor, 0.3-4g/L potassium dihydrogen phosphate, 0.2-3g/L magnesium sulfate, 5-30g/L seawater crystal, 5-20 mg/L vitamin B13, 20-20 mg/L vitamin B63, 10-10 mg/L vitamin B121 and 1-10mg/L biotin.
In the fermentation process, sugar supplement is adopted, the glucose concentration in the system is controlled to be between 5 and 10g/L, and the glucose concentration is not higher than 3g/L after the fermentation is finished.
A single colony of fresh schizochytrium SC01 strain is picked up and cultured in 50ml of seed culture medium under the condition of shaking (180rpm) at 25 ℃ for 90 hours, and the biomass, DHA content and astaxanthin content are respectively measured.
And (3) detecting the DHA content: putting 50mg of dried bacterial powder into 0.4M KOH/methanol solution, carrying out ultrasonic crushing, and carrying out water bath at 60 ℃ for 1 hour; then, 14% BF 3/methanol solution was added and esterified at 60 ℃ for 1 hour. The fatty acid methyl ester was extracted with n-hexane and then analyzed by gas chromatography. Fatty acid methyl esters were analyzed by gas chromatography (Agilent 7890B) equipped with a FID detector and a DB-23 capillary column (30 m.times.0.25 mm). Nitrogen gas is used as carrier gas, the initial column temperature is 100 ℃, after 1 minute, the temperature is raised to 250 ℃ at the speed of 15 ℃/min, and the temperature is kept for 10 minutes. The temperature of the sample inlet is 250 ℃, and the temperature of the detector is 260 ℃; the sample amount is 1 ul; the DHA was quantified by comparison to a pre-established DHA standard sample concentration curve.
Detection of astaxanthin: taking 50mg of bacterial powder, adding liquid nitrogen, grinding for 5min by using a mortar for cell wall breaking, adding 3mL of acetone, carrying out vortex oscillation for 15s, placing in a 50 ℃ water bath for 30min, oscillating for 15s every 10min, centrifuging at 3500rpm for 10min, taking supernatant, adding 2mL of acetone, carrying out vortex oscillation for 15s, centrifuging again, and repeating the operations until the thallus turns white. Drying the astaxanthin acetone extracting solution by using nitrogen, metering the volume by using 25mL of methanol, and storing the astaxanthin acetone extracting solution in a refrigerator at the temperature of 20 ℃ below zero for later use.
Carrying out quantitative detection on astaxanthin by using a high performance liquid chromatography: Nova-Pak C18 column (150 mm. times.3.9 mm, 5 μm, Waters corporation, USA); the mobile phase A is water, and the mobile phase B is methanol; elution gradient: 10% A, 90% B (0 min); 10% A, 90% B (1 min); 0% A, 100% B (10 min); 0% A, 100% B (20 min). Flow rate 1 mL/min-1(ii) a The detector is a Waters 996 photodiodeA tube array detector; the spectrum scanning wavelength range is 300-700 nm, and the detection wavelength is 476 nm; the amount of sample was 10. mu.L.
The content of DHA in the schizochytrium SC01 cells was determined to be 26% and the content of astaxanthin to be 0.34% (see Table 1).
TABLE 1 content of DHA and astaxanthin in biomass of Schizochytrium SC01 (percentage on dry basis)
Figure BDA0001485106600000051
A food contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
A food additive contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
A health product contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
A feed additive contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
The schizochytrium limacinum adopting the technical scheme and the application thereof have the following advantages:
(1) the invention provides a schizochytrium capable of simultaneously producing DHA and astaxanthin with high yield under the condition of no illumination: aurantiochytrium sp.SC01 with preservation number of CGMCC No. 14574; the strain can grow rapidly at the temperature of 20-30 ℃, and a large amount of DHA and astaxanthin are accumulated.
(2) The growth speed is high, and the maximum yield can be obtained within 80-100 hours under the fermentation condition; after 100 hours of fermentation, the biomass is 70-120g/L, DHA accounts for 24-26% of the dry weight of the cells, and the astaxanthin content accounts for 0.3-0.5% of the dry weight of the cells.
(3) The yield of the DHA in the biological oil is greatly improved, the production cost of the DHA can be greatly reduced, and the industrial process of producing the DHA by fermenting marine microorganisms is further promoted.
Drawings
FIG. 1 is a view under a morphological optical microscope under a microscope of a Schizochytrium limacinum strain of the present invention;
FIG. 2 is a scanning electron microscope image of the Schizochytrium limacinum strain of the present invention.
Detailed Description
The schizochytrium limacinum is classified and named as schizochytrium sp, and the laboratory is named as schizochytrium limacinum SC01 strain, and has been preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.14574 and the preservation date of 2017, 8 and 29 days.
The Schizochytrium limacinum strain is obtained by separating kelp residue collected from Shandong Ronghai area by a pine pollen fishing method. Observed by an optical microscope, the following results are found: the thallus is spherical or ellipsoidal, has a diameter of 8-22 microns, is orange red, and is darkened along with the prolonging of the culture time, and has double flagella with different lateral lengths, and intracellular fatty acids mainly comprise hexadecanoic acid, octadecanoic acid, arachidonic acid, docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA), wherein the DHA content accounts for more than 50% of the total fatty acids, and the astaxanthin content is about 0.3%. Comparing the 18S rDNA sequence by a molecular biology means, finding that the homology of the strain and the 18S rDNA sequence of the schizochytrium is more than 99 percent, thereby judging that the strain belongs to the schizochytrium sp.SC01 and being named as Aurantiochytrium sp.SC01 in a laboratory.
The carbon source which can be utilized by the schizochytrium sp.SC01 strain comprises glucose, molasses, fructose and glycerol, the most suitable carbon source is glucose, and the concentration is 30g/L-80 g/L; both organic and inorganic nitrogen sources can be used, wherein the organic nitrogen source comprises yeast extract, corn steep liquor, peptone and urea; the inorganic nitrogen source comprises nitrate and ammonium salt, the most suitable nitrogen source is yeast extract, and the concentration is 5g/L-35 g/L. In addition, corn steep liquor is also a suitable nitrogen source, and the concentration is 1g/L-5 g/L.
The optimum growth temperature of the schizochytrium sp.SC01 strain is 20-30 ℃, and the maximum yield of astaxanthin and DHA can be obtained after 96 hours of fermentation.
Example 1
The 3L fermentor culture with SC01 strain included the following steps:
a. inoculating the strain stored in Glycine max (L.) Gaertn tube into a 250ml shake flask containing 50ml seed culture medium, and culturing in a shaker at 30 deg.C at 200rpm for 24 hr to obtain first-stage seed;
b. inoculating the primary seeds into a 500ml shake flask filled with 100ml of seed culture medium, and culturing for 12h in a shaking table at 30 ℃ at the rotating speed of 200rpm to obtain secondary seeds;
c. 1.2L of fermentation medium was added to a 3L bioreactor and the activated secondary seed liquid was inoculated.
Wherein: the seed culture medium contains 10g/L of glucose, 5g/L of yeast extract and 30g/L of seawater crystal;
the fermentation medium contains 80g/L glucose, 35g/L yeast extract, 5g/L corn steep liquor, 4g/L potassium dihydrogen phosphate, 3g/L magnesium sulfate, 30g/L crystal, and vitamin B120mg/L, vitamin B620mg/L, vitamin B1210mg/L and biotin 10 mg/L.
During the fermentation process, the temperature is controlled at 30 ℃. In the fermentation process, nitrogen supplement operation is adopted, 20% (w/v) yeast powder solution is added in the fermentation within 36-64 hours, 2M NaOH is automatically added to keep the pH value at 6.5, and the fermentation time is 80 hours.
Detecting the yield of DHA and astaxanthin: the yield of Schizochytrium obtained in the 3L fermentor (see Table 2) was 90.34g/L, DHA was 23.4g/L and astaxanthin was 0.272 g/L.
TABLE 2 fatty acid composition in cells of Schizochytrium SC01 after fermentation
Figure BDA0001485106600000081
Example 2
The 10L fermentor culture with SC01 strain includes the following steps:
a. inoculating the strain stored in Glycine max (L.) Gaertn tube into a 250ml shake flask containing 50ml seed culture medium, and culturing in a shaking table at 20 deg.C at 200rpm for 24 hr to obtain first-stage seed;
b. inoculating the primary seeds into a 500ml shake flask filled with 100ml of seed culture medium, and culturing for 12h in a shaking table at 20 ℃ at the rotating speed of 200rpm to obtain secondary seeds;
c. 6L of fermentation medium was added to a 10L bioreactor and the activated secondary seed liquid was inoculated.
Wherein: the seed culture medium contains 40g/L of glucose, 15g/L of yeast extract and 5g/L of seawater crystal;
the fermentation medium contains 20g/L glucose, 5g/L yeast extract, 1g/L corn steep liquor, 0.3g/L potassium dihydrogen phosphate, 0.2g/L magnesium sulfate, 5g/L seawater crystal, vitamin B13 mg/L, vitamin B63 mg/L, vitamin B121 mg/L and biotin 1 mg/L.
During the fermentation, the temperature was controlled at 20 ℃. In the fermentation process, nitrogen supplement operation is adopted, and 20% (w/v) of corn steep liquor solution is fed in within 36-64 hours of fermentation. The pH was maintained at 6.5 by automatic addition of 2M NaOH. The fermentation time was 100 hours.
After the fermentation is finished, the yield of the schizochytrium limacinum obtained in a 10L fermentation tank is 70.25g/L, the yield of the astaxanthin obtained is 0.365g/L, and the yield of the DHA obtained is 18.4 g/L.
Example 3
The 100L fermentor culture with SC01 strain includes the following steps:
a. inoculating the strain stored in Glycine max (L.) Gaertn tube into a 250ml shake flask containing 50ml seed culture medium, and culturing in a shaking table at 20 deg.C at 200rpm for 24 hr to obtain first-stage seed;
b. inoculating the primary seeds into a 500ml shake flask filled with 100ml of seed culture medium, and culturing for 12h in a shaking table at 20 ℃ at the rotating speed of 200rpm to obtain secondary seeds;
c. 60L of fermentation medium was added to a 100L bioreactor and 10L of activated seed liquid was inoculated.
Wherein: the seed culture medium contains 40g/L of glucose, 15g/L of yeast extract and 5g/L of seawater crystal;
the fermentation medium contains 60g/L glucose, 15g/L yeast extract, 5g/L corn steep liquor, 4g/L potassium dihydrogen phosphate, 2g/L magnesium sulfate, 10g/L seawater crystal, 110 mg/L vitamin B, 610 mg/L vitamin B, 123 mg/L vitamin and 5mg/L biotin.
During the fermentation, the temperature was controlled at 23 ℃. In the fermentation process, nitrogen supplement operation is adopted, and 20% (w/v) yeast powder solution is fed in within 36-64 hours of fermentation. The pH was maintained at 6.5 by automatic addition of 2M NaOH. The fermentation time was 96 hours.
After the fermentation is finished, the yield of the schizochytrium limacinum obtained in a 100L fermentation tank is 119.86g/L, the yield of the obtained DHA is 30.8g/L, and the yield of the obtained astaxanthin is 0.528 g/L.
Example 4
A food additive contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
Example 5
A food, wherein the food contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
Example 6
A health product contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
Example 7
A feed additive, wherein the feed additive contains Schizochytrium limacinum with the preservation number of CGMCC No. 14574.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, it is possible to make various changes and modifications without departing from the structure of the present invention, and these should be considered as the protection scope of the present invention, which will not affect the effect of the implementation of the present invention and the utility of the patent.

Claims (7)

1. A Schizochytrium limacinum, which is characterized in that: the classification of the strain is named as schizochytrium limacinum
(Aurantiochytrium sp.) SC01, which is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms with the preservation number of CGMCC No. 14574.
2. Use of the schizochytrium limacinum of claim 1 for the preparation of docosahexaenoic acid (DHA).
3. Use of the schizochytrium limacinum of claim 1 in the preparation of astaxanthin.
4. A food product characterized by: the food contains schizochytrium with the preservation number of CGMCC No. 14574.
5. A health product is characterized in that: the health product contains Schizochytrium limacinum with preservation number of CGMCC No. 14574.
6. A food additive characterized by: the food additive contains schizochytrium with the preservation number of CGMCC No. 14574.
7. A feed additive, characterized in that: the feed additive contains schizochytrium with the preservation number of CGMCC No. 14574.
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