公开(公告)号：US11414650B2

公开(公告)日：2022-08-16

申请号：US17120163

申请日：2020-12-12

Applicant: SHANDONG UNIVERSITY OF TECHNOLOGY

Inventor： Yuanda Song ,  Md. Ahsanul Kabir Khan ,  Junhuan Yang ,  Huaiyuan Zhang ,  Wu Yang ,  Qing Liu

IPC: C12N9/02 ,  C12P7/64 ,  C12P7/6463 ,  C12N1/14 ,  C12N15/80

Abstract: 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.

公开(公告)号：US11479778B2

公开(公告)日：2022-10-25

申请号：US17285816

申请日：2020-11-30

Applicant: SHANDONG UNIVERSITY OF TECHNOLOGY

Inventor： Yuanda Song ,  Caili Sun ,  Huaiyuan Zhang ,  Wu Yang ,  Meiling Chen

IPC: C12N15/80 ,  C12N9/10

Abstract: The present invention relates to the technical field of gene engineering and particularly relates to a method for constructing non-de novo synthesized Mucor circinelloides recombinant strain with high lipid yield, recombinant strain constructed by method, and application of recombinant strain. According to the present invention, a diacylglycerol acyltransferase gene (DGAT) is overexpressed in Mucor circinelloides WJ11 by a homologous recombination technology, and exogenous oil is added for fermentation, such that the non-de novo synthesized Mucor circinelloides recombinant strain with high lipid yield is constructed. Compared with the control strain Mc2075, the fat yield of the Mucor circinelloides is increased; and when the diacylglycerol acyltransferase (DGAT) is transformed into the uracil defective type of Mucor circinelloides WJ11, the fatty acid composition changes after fermentation, and the lipid content may reach 53% of dry cell weight after the fermentation condition is optimized.