The present disclosure relates to a construction method of a Mucor circinelloides cell factory for producing dihomo-γ-linolenic acid and a fermentation technology, belonging to the field of genetic engineering. In the present disclosure, γ-linolenic acid elongase gene glelo is obtained from Mortierella alpine by cloning, the gene is ligated to an integrative plasmid pMAT1552, and transformed into a Mucor circinelloides defective strain Mu402, and the gene glelo is integrated into Mucor circinelloides genome through homologous recombination, to obtain the recombinant strain Mc-glelo, and finally, the expression of the gene glelo in Mucor circinelloides is realized. The lipid content in the recombinant strain Mc-glelo is not obviously different from that in the control strain Mc1552, however, the lipid composition changes greatly, and dihomo-γ-linolenic acid appears in the lipids of the recombinant strain Mc-glelo, and the content thereof reaches 5.7% of the total fatty acids. Under optimized fermentation conditions and in the presence of precursor fatty acid, the DGLA content reaches 7.6%. The new recombinant strain was deposited in China General Microbiological Culture Collection Center on Jun. 20, 2018, with the address of No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. The accession number given to the biological material by the collection center is CGMCC No. 15887, and the suggested taxonomic denomination is Mucor circinelloides-GLELO.

A method of producing lipids, containing the steps of: culturing a transformant into which a gene encoding at least one of the proteins selected from the group consisting of the following proteins (A) to (C) is introduced; and producing fatty acids or lipids containing the same as components:
(A) A protein consisting of the amino acid sequence of the 23.sup.rd to 146.sup.th amino acids set forth in SEQ ID NO: 1;
(B) A protein consisting of an amino acid sequence having 70% or more identity with the amino acid sequence of the protein (A), and having acyl carrier protein activity; and
(C) A protein containing the amino acid sequence of the protein (A) or (B), and having acyl carrier protein activity.

Provided herein are methods of culturing a microorganism. The methods include providing a container comprising one or more microorganisms in a medium, which has a first carbon to nitrogen ratio; culturing the microorganisms until the culture reaches a threshold indicator; harvesting a portion of the culture while maintaining the majority of the culture in the container; and adding fresh medium comprising a second carbon to nitrogen ratio to the container with the majority of the culture comprising the microorganisms.

Provided herein are methods of culturing a microorganism. The methods include providing a container comprising one or more microorganisms in a medium, which has a first carbon to nitrogen ratio; culturing the microorganisms until the culture reaches a threshold indicator; harvesting a portion of the culture while maintaining the majority of the culture in the container; and adding fresh medium comprising a second carbon to nitrogen ratio to the container with the majority of the culture comprising the microorganisms.

The present invention provides a process for the production of a microbial oil comprising culturing a micro-organism in a two stage fermentation process where, in a last stage that precedes the end of fermentation, the carbon source is: consumed by the micro-organisms at a rate greater than it is added to the medium; added at a rate #0.30 M carbon/kg medium; or is rate limiting on the growth of the micro-organism. The micro-organisms thus have the carbon source restricted so that they preferentially metabolise fats or lipids other than arachidonic acid (ARA), so increasing the proportion of ARA in the cells. A microbial oil is then recovered from the micro-organism, using hexane as a solvent, that has at least 50% ARA and at least 90% triglycerides.

The invention provides isolated nucleic acid molecules which encode novel fatty acid desaturase family members. The invention also provides recombinant expression vectors containing desaturase nucleic acid molecules, host cells into which the expression vectors have been introduced, and methods for large-scale production of long chain polyunsaturated fatty acids (LCPUFAs), e.g., DHA.

The present invention relates to a process for the production of polyunsaturated fatty acids in an organism by introducing, into the organism, nucleic acids which encode polypeptides with ?5-elongase activity. Advantageously, these nucleic acids can be expressed in the organism together with further nucleic acids which encode polypeptides of the biosynthesis of the fatty acid or lipid metabolism. Especially advantageous are nucleic acids which encode ?6-desaturases, ?5-desaturases, ?4-desaturases and/or ?6-elongases. These desaturases and elongases are advantageously derived from Thalassiosira, Euglena or Ostreococcus. The invention furthermore relates to a process for the production of oils and/or triacylglycerides with an elevated content of long-chain polyunsaturated fatty acids, and oils and/or triacylglycerides thus obtained. The invention also relates to the nucleic acids, and constructs, vectors and transgenic organisms comprising the same, as well as oils, lipids and/or fatty acids produced by the process according to the invention and to their use.

The present invention relates to a process for the production of polyunsaturated fatty acids in an organism by introducing, into the organism, nucleic acids which encode polypeptides with 5-elongase activity. Advantageously, these nucleic acids can be expressed in the organism together with further nucleic acids which encode polypeptides of the biosynthesis of the fatty acid or lipid metabolism. Especially advantageous are nucleic acids which encode 6-desaturases, 5-desaturases, 4-desaturases and/or 6-elongases. These desaturases and elongases are advantageously derived from Thalassiosira, Euglena or Ostreococcus. The invention furthermore relates to a process for the production of oils and/or triacylglycerides with an elevated content of long-chain polyunsaturated fatty acids, and oils and/or triacylglycerides thus obtained. The invention also relates to the nucleic acids, and constructs, vectors and transgenic organisms comprising the same, as well as oils, lipids and/or fatty acids produced by the process according to the invention and to their use.

The current invention provides use of a CLA-producing bacterium for the in vivo conversion in the gut of polyunsaturated fatty acids to CLA. The CLA-producing bacterium is selected from one or more of the group consisting of propionibacteria, lactobacilli, lactococci and streptococci, and bifidobacteria.

The present invention provides a production method of oxo fatty acid, as well as rare fatty acids such as conjugated fatty acid, hydroxylated fatty acid, partially saturated fatty acid and the like, which uses 4 kinds of enzymes (fatty acid-hydratase, hydroxylated fatty acid-dehydrogenase, oxo fatty acid-isomerase, oxo fatty acid-enone reductase) derived from Lactobacillus plantarum including lactic acid bacteria and the like. Furthermore, the present invention also provides a more efficient production method of oxo fatty acid and the like, which partly uses a chemical oxidation reaction in combination.