Fungal cells for tailored fats

1 . A fungal cell capable of producing triacylglycerols with fatty acids with an acyl chain having a target characteristic, wherein

the fungal cell is genetically modified for expression of a heterologous lysophosphatidyl acyltransferase (LPAT) (EC 2.3.1.51) capable of esterifying a triacylglycerol precursor with an acyl-CoA with an acyl chain having the target characteristic; and

the heterologous LPAT has preference for esterifying a triacylglycerol precursor with a saturated fatty acyl-CoA over esterifying a triacylglycerol precursor with an unsaturated fatty acyl-CoA,

wherein the heterologous LPAT is selected from the group consisting of Bos taurus AGPAT4 as defined in SEQ ID NO: 13, 14, or 15, Bos taurus AGPAT1 as defined in SEQ ID NO: 16, Bos taurus AGPAT2 as defined in SEQ ID NO: 17, Bos taurus AGPAT3 as defined in SEQ ID NO: 18, Bos taurus AGPAT5 as defined in SEQ ID NO: 19, Arabidopsis thaliana LPAT1 as defined in SEQ ID NO: 20, Brassica napus LPAT1 as defined in SEQ ID NO: 24 , Mycolicibacterium smegmatis ERS451418_00313 as defined in SEQ ID NO: 28 , Mycolicibacterium smegmatis Probable LPAT as defined in SEQ ID NO: 29 , Mycolicibacterium smegmatis ERS451418_06226 as defined in SEQ ID NO: 30 , Mycolicibacterium smegmatis ERS451418_02370 as defined in SEQ ID NO: 31 , Mycolicibacterium smegmatis ERS451418_05575 as defined in SEQ ID NO: 32 , Mycolicibacterium smegmatis ERS451418_04128 as defined in SEQ ID NO: 33 , Mycolicibacterium smegmatis ERS451418_06227 as defined in SEQ ID NO: 34 , Mycolicibacterium smegmatis BIN_B_00519 as defined in SEQ ID NO: 35 , Mycolicibacterium smegmatis D806_035910 as defined in SEQ ID NO: 36 , Mycolicibacterium smegmatis BIN_B_03706 as defined in SEQ ID NO: 37 Mycolicibacterium smegmatis D806_003290 as defined in SEQ ID NO: 38, Cocos nucifera probable LPAT as defined in SEQ ID NO: 86, and a heterologous LPAT having at least 95% sequence identity to any of SEQ ID NOs: 13-20, 24, 28-38, or 86.

2 . The fungal cell according to claim 1 , wherein

the fungal cell is genetically modified for overexpression of a triacylglycerol lipase (EC 3.1.1.3); and

the triacylglycerol lipase has higher lipase activity on unsaturated fatty acids of triacylglycerols compared to saturated fatty acids.

3 . The fungal cell according to claim 2 , wherein the triacylglycerol lipase is selected from the group consisting of Homo sapiens PNLIP as defined in SEQ ID NO: 11, Yarrowia lipolytica Lip2 as defined in SEQ ID NO: 12 , Diutina rugosa LIP1 as defined in SEQ ID NO: 39, Streptomyces rimosus CP984 RS32550 as defined in SEQ ID NO: 40, Geotrichum candidum LIP1 as defined in SEQ ID NO: 41, Geotrichum candidum LIP2 as defined in SEQ ID NO: 42, Streptomyces coelicolor LIP1 as defined in SEQ ID NO: 43 , Amycolatopsis mediterranei AMED 3680 as defined in SEQ ID NO: 45, Penaeus vannamei C7M84_014708 as defined in SEQ ID NO: 48, Solanum lycopersicum LeLID1 as defined in SEQ ID NO: 50 , Pseudozyma aphidis LIPA as defined in SEQ ID NO: 51, and a triacylglycerol lipase having at least 70% sequence identity to any of SEQ ID NOs: 11-12, 39-43, 45, 48, or 50-51.

4 . The fungal cell according to claim 1 , wherein the fungal cell is genetically modified for expression of

(a) a heterologous glycerol-3-phosphate acyltransferase (GPAT) selected from the group consisting of Bos taurus GPAT4 isoform X1 as defined in SEQ ID NO: 52, Bos taurus GPAT4 isoform X2 as defined in SEQ ID NO: 53, Bos taurus GPAT2 as defined in SEQ ID NO: 54, Bos taurus GPAT3 isoform X1 as defined in SEQ ID NO: 55, Bos taurus GPAT3 isoform X2 as defined in SEQ ID NO: 56, Bos taurus GPAT3 isoform X3 as defined in SEQ ID NO: 57, Bos taurus GPAT3 isoform X4 as defined in SEQ ID NO: 58, Mus musculus GPAT4 isoform X1 as defined in SEQ ID NO: 59, Homo sapiens GPAM isoform X1 as defined in SEQ ID NO: 60, Cucurbita moschata ATS1;2 as defined in SEQ ID NO: 61, Mus musculus GPAT2 as defined in SEQ ID NO: 63, and a heterologous GPAT having at least 70% sequence identity to any of SEQ ID NOs: 52-61, or 63; and/or

(b) a heterologous diacylglycerol acyltransferase (DGAT) selected from the group consisting of Brassica napus DGAT1-1 as defined in SEQ ID NO: 7, Tropaeolum majus DGAT as defined in SEQ ID NO: 10, Bos taurus DGAT1 isoform X1 as defined in SEQ ID NO: 64, Bos taurus DGAT1 isoform X2 as defined in SEQ ID NO: 65, Bos taurus DGAT2 as defined in SEQ ID NO: 66, Bos taurus DGAT2L6 as defined in SEQ ID NO: 67, Homo sapiens DGAT2 as defined in SEQ ID NO: 69, Arachis hypogaea DGAT3 as defined in SEQ ID NO: 70 , Arabidposis thaliana DGAT1 as defined in SEQ ID NO: 71 , Thraustochytrium aureum DGAT2 as defined in SEQ ID NO: 72, and a heterologous DGAT having at least 70% sequence identity to any of SEQ ID NOs: 7, 10, 64-67, or 69-72.

5 . The fungal cell according to claim 1 , wherein the fungal cell is genetically modified for:

a) downregulation or deletion of

i) an endogenous acyl-CoA oxidase;

ii) an endogenous glycerol-3-phosphate acyltransferase;

iii) an endogenous lysophosphatidyl acyltransferase;

iv) an endogenous diacylglycerol acyltransferase;

v) an endogenous desaturase; and/or

vi) an endogenous lipase; and/or

b) enhanced activity of an acetyl-CoA carboxylase.

6 . A method for producing triacylglycerols comprising:

culturing a fungal cell according to claim 1 in a culture medium and in culture conditions suitable for production of the triacyclglycerols by the fungal cell, preferably nitrogen-limited conditions; and

collecting the triacyclglycerols from the culture medium and/or the fungal cell.