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1. (WO2019048327) METHOD OF SEPARATING LIPIDS FROM A LYSED LIPIDS CONTAINING BIOMASS
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A method of separating a polyunsaturated fatty acids (PUFAs) containing lipid from the debris of a biomass, comprising the following steps:

a) Providing a suspension of a biomass comprising cells which contain a PUFAs containing lipid;

b) Lysing the cells of the biomass;

c) Adding to the suspension as obtained in step (b) acetone, until a final amount of

between 25 and 47.5 wt.-% of acetone is reached;

d) Thoroughly mixing the suspension as obtained in step (c);

e) Separating the oil and acetone containing light phase as obtained in step (d) from the water, acetone, salt and cell debris containing heavy phase.

The method according to claim 1, wherein acetone is added to the suspension of biomass in step (c), until a final amount of between 27.5 and 45.0, in particular 30.0 to 42.5, preferably 30.0 to 40.0 wt.-% of acetone is reached.

The method according to any of the preceding claims, wherein mixing of the suspension in step (d) is carried out by shaking, stirring and/or vortexing.

The method according to any of the preceding claims, wherein lysing of the cells of the biomass is carried out enzymatically, mechanically, chemically and/or physically.

The method according to claim 4, wherein lysing of the cells of the biomass comprises an enzymatic treatment of the cells with a cell-wall degrading enzyme.

The method according to claim 5, wherein lysing of the cells of the biomass is carried out as follows:

i) Heating the suspension of the biomass to a temperature of between 50°C and 70°C, preferably to a temperature of between 55°C and 65°C, adding a cell wall-degrading enzyme to the fermentation broth, and adjusting an adequate pH value, if necessary, at which the enzyme is properly working;

ii) Keeping the temperature and pH in the ranges as depicted in (i) for at least one hour, preferably for at least two hours, more preferably for two to four hours.

7. The method according to any of the preceding claims, wherein after lysing of the cells, the suspension is concentrated to a total dry matter content of 30 to 60 wt.-%, more preferably 35 to 55 wt.-%, in case that the suspension has a lower TDM content.

8. The method according to any of the preceding claims, wherein steps (c) to (e) are carried out at a temperature of 10 to 50°C, preferably 15 to 40°C, more preferably 18 to 35°C, in particular at about room temperature.

9. The method according to any of the preceding claims, wherein before addition of acetone according to step (c) an acidic pH value is adjusted, in particular a pH value of 2.5 to 6.8, preferably a pH value of 3.0 to 6.0, in case that the suspension has a different pH value.

10. The method according to any of the preceding claims, wherein separation of the oil and acetone containing light phase from the water, acetone, salt and cell debris containing heavy phase is realized by mechanical means and preferably at a pH value of 5.5 to 8.5, more preferably 6.0 to 8.0.

11. The method according to any of the preceding claims, comprising as further step the

separation of the acetone from the PUFAs containing oil.

12. The method according to any of the preceding claims, wherein the suspension has a biomass density at least 80, 100, 120 or 140 g/l.

13. The method according to any of the preceding claims, wherein the cells which contain a PUFAs containing lipid are selected from algae, fungi, protists, bacteria, microalgae, plant cells, and mixtures thereof.

14. The method according to claim 12, wherein the microalgae are selected from the phylus Stramanopiles, in particular of the family of Thraustochytrids, preferably of the genus Schizochytrium.

15. The method according to any of claims 5 to 13, wherein the cell-wall degrading enzyme is selected from proteases, cellulases, hemicellulases, chitinases, pectinases, sucrases, maltases, lactases, alpha-glucosidases, beta-glucosidases, amylases, lysozymes,

neuraminidases, galactosidases, alpha-mannosidases, glucuronidases, hyaluronidases,

pullulanases, glucocerebrosidases, galactosylceramidases, acetylgalactosaminidases, fucosidases, hexosaminidases, iduronidases, maltases-glucoamylases, beta-glucanases, mannanases, and combinations thereof.