Construction of Mucor circinelloides cell factory for producing stearidonic acid and fermentation technology thereof

Patent metadata

Field	Value
Publication number	US-11345937-B2
Application number	US-202017120160-A
Country	US
Kind code	B2
Filing date	Dec 12, 2020
Priority date	Dec 11, 2018
Publication date	May 31, 2022
Grant date	May 31, 2022

Abstract

Provided is construction of Mucor circinelloides cell factory for producing stearidonic acid (SDA) and fermentation technology. Δ15 Desaturase gene is obtained by cloning from Mortierella alpina , the gene is ligated to an integrative plasmid pMAT1552, and transformed into a Mucor circinelloides defective strain Mu402, and Δ15 Desaturase gene is integrated on Mucor circinelloides genome through homologous recombination, to obtain the recombinant strain Mc-Δ15, finally, the expression of the Δ15 Desaturase gene in Mucor circinelloides is realized. The recombinant new strain is accession number CGMCC No. 15888, and the classification name is Mucor circinelloides -D15D.

First claim

What is claimed is:

1. A recombinant cell for producing stearidonic acid, wherein the recombinant cell comprises a polynucleotide encoding Δ15 desaturase, and the polynucleotide encoding Δ15 desaturase is operably ligated to a promoter that directs the polynucleotide to be expressed in a cell,
wherein the polynucleotide encoding Δ15 desaturase comprises the polynucleotide sequence set forth in SEQ ID NO: 1, and
wherein the recombinant cell is obtained from a Mucor cell.

2. The recombinant cell of claim 1 , wherein the recombinant cell is obtained from a Mucor circinelloides cell.

3. A method for preparing a recombinant cell for producing stearidonic acid, the method comprising:
introducing into the cell a polynucleotide encoding Δ15 desaturase,
wherein the polynucleotide encoding the Δ15 desaturase is operably ligated to a promoter that directs the polynucleotide to be expressed in the cell,
wherein the polynucleotide encoding Δ15 desaturase comprises
the polynucleotide sequence set forth in SEQ ID NO: 1, and
wherein the recombinant cell is obtained from a Mucor cell.

4. A method for producing stearidonic acid in a recombinant cell, comprising:
introducing into the recombinant cell a polynucleotide encoding Δ15 desaturase,
wherein the polynucleotide encoding Δ15 desaturase is operably ligated to a promoter that directs the polynucleotide to be expressed in the cell; and
fermenting the recombinant cell to produce stearidonic acid,
wherein the polynucleotide encoding Δ15 desaturase com prises the polynucleotide sequence set forth in SEQ ID NO: 1; and
wherein the recombinant cell is obtained from a Mucor cell.

5. The method of claim 4 , wherein the fermenting is carried out in a fermentation culture medium, and the fermentation culture medium comprises linolenic acid, linolenic acid compounds and/or precursor compounds of linolenic acid.

6. The method of claim 5 , wherein the fermentation culture medium comprises γ-linolenic acid and/or γ-linolenic acid compounds.

7. The method of claim 5 , wherein the linolenic acid and/or the linolenic acid compounds are in a vegetable oil.

8. The method of claim 7 , wherein the vegetable oil is a safflower seed oil and/or a sunflower seed oil.

9. The method of claim 7 , wherein the vegetable oil is initially present in the fermentation culture medium, or is added to the fermentation culture medium as a supplement during fermenting.

Assignees

• Univ Shandong Technology

Inventors

• Song Yuanda
• Khan Md Ahsanul Kabir
• Yang Junhuan
• Zhang Yao
• Yang Wu
• Li Shaoqi

Classifications

• C12N9/0071 Primary

  acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title

• C12N9/0004

  Oxidoreductases (1.) · CPC title

• C12P7/6427 Primary

  Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone · CPC title