# US 2021/0340583 A1 · Schizochytrium strain and use thereof, microbial oil containing dha at sn-2 position and preparation and use thereof

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- Number: US 2021/0340583 A1
- Document ID: US20210340583A1
- Status: Published application
- Kind: A1
- Application number: 17/375,551
- Filed: 2021-07-14
- Published: 2021-11-04
- Earliest claimed: 2019-11-26
- Assignees: Hanpeng QU
- Inventors: Yun LIANG; Sheng CAO; Shenjian WANG
- Web page: https://patentgrep.com/patent/US20210340583A1

## CPC

- A23L 33/115: Fatty acids or derivatives thereof; Fats or oils
- C12P 7/6463: obtained from glyceride producing microorganisms, e.g. single cell oil
- C12N 1/12: Unicellular algae; Culture media therefor (as new plants A01H13/00)
- C12N 1/125: Unicellular algae isolates
- C12N 1/14: Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor
- C12P 7/6434: Docosahexenoic acids [DHA]
- C12P 7/6472: containing polyunsaturated fatty acid [PUFA] residues, i.e. having two or more double bonds in their backbone
- C12R 2001/89: Algae {; Processes using algae}
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

This disclosure relates to microbial technology, and more particularly to a Schizochytrium strain and a use thereof, a microbial oil containing DHA at an Sn-2 position, and a preparation and uses thereof. Sn-2 fatty acids of a triglyceride in the microbial oil contain 23% or more by weight of DHA. The microbial oil is prepared by fermentation using a Schizochytrium strain, where the Schizochytrium strain is Schizochytrium sp. with an accession number of GDMCC No. 60733.

## Claims

### Claim 1

A microbial oil, comprising:
a triglyceride;
wherein Sn-2 fatty acids of the triglyceride comprises 23% or more by weight of DHA.

### Claim 2 (depends on 1)

The microbial oil of claim 1, wherein a weight percentage of total DHA in the microbial oil is not less than 38%.

### Claim 3

A method for producing a microbial oil, comprising:
inoculating a Schizochytrium strain into a fermentation medium for fermentation to produce the microbial oil;
wherein the Schizochytrium strain is Schizochytrium sp. GDMCC No. 60733;
the fermentation is carried out at pH 6-7.5 and 27-31° C. for 4-8 days under a ventilation rate of 0.45-1.1 vvm;
the fermentation medium comprises 5-70 g/L of a carbon source, 15-45 g/L of a nitrogen source, 5-40 g/L of an inorganic salt, 0.01-0.04 g/L of a trace element and 0.01-0.04 g/L of a vitamin;
the nitrogen source comprises glutamate; and a concentration of glutamate in the fermentation medium is 5-15 g/L.

### Claim 4 (depends on 3)

The method of claim 3, wherein the method comprises:
inoculating the Schizochytrium sp. GDMCC No. 60733 into an activation medium for activation to obtain an activated Schizochytrium suspension;
inoculating the activated Schizochytrium suspension into a seed culture medium for expansion culture to obtain a seed liquid; and
inoculating the seed liquid into the fermentation medium for the fermentation to produce the microbial oil;
wherein the activation is carried out at 27.5-28.5° C. and 150-200 r/min for 48-72 hours;
the activation medium comprises 30-50 g/L of a carbon source, 25-45 g/L of a nitrogen source, 25-40 g/L of an inorganic salt, 0.015-0.025 g/L of a trace element and 0.01-0.02 g/L of a vitamin;
the expansion culture is carried out at 27-28.5° C. under a ventilation rate of 0.5-0.8 vvm for 48-72 hours; and
the seed culture medium comprises 25-60 g/L of a carbon source, 10-30 g/L of a nitrogen source, 15-35 g/L of an inorganic salt, 0.01-0.02 g/L of a trace element and 0.01-0.02 g/L of a vitamin.

### Claim 5 (depends on 4)

The method of claim 4, wherein the carbon source in the fermentation medium, the carbon source in the activation medium and the carbon source in the seed culture medium are independently selected from the group consisting of glucose, sucrose and a combination thereof;
the nitrogen source in the fermentation medium, the nitrogen source in the activation medium and the nitrogen source in the seed culture medium are independently selected from the group consisting of sodium glutamate, yeast powder, yeast extract and a combination thereof;
the inorganic salt in the fermentation medium, the inorganic salt in the activation medium and the inorganic salt in the seed culture medium are independently selected from the group consisting of calcium salt, phosphate, potassium salt, sodium salt, magnesium salt, ammonium salt and a combination thereof;
the trace element in the fermentation medium, the trace element in the activation medium and the trace element in the seed culture medium are independently selected from the group consisting of nickel, copper, molybdenum, cobalt, zinc, iron, manganese and a combination thereof; and/or
the vitamin in the fermentation medium, the vitamin in the activation medium and the vitamin in the seed culture medium are independently selected from the group consisting of vitamin B1, vitamin B12, vitamin B6, calcium pantothenate, biotin and a combination thereof.

### Claim 6 (depends on 3)

The method of claim 3, further comprising:
subjecting a product of the fermentation to extraction to produce the microbial oil.

### Claim 7 (depends on 1)

A food comprising the microbial oil of claim 1, wherein the food is an infant formula food, a nutraceutical or a health food.

### Claim 8

A Schizochytrium strain, wherein the Schizochytrium strain has an accession number of GDMCC No. 60733.

### Claim 9 (depends on 8)

A method of preparing a microbial oil using the Schizochytrium strain of claim 8, comprising:
inoculating the Schizochytrium strain into a fermentation medium for fermentation to produce the microbial oil;
wherein the microbial oil comprises a triglyceride;
Sn-2 fatty acids of the triglyceride comprise 23% or more by weight of DHA; and
a weight percentage of total DHA in the microbial oil is not less than 38%.