In situ two-phase extraction system

Patent metadata

Field	Value
Publication number	US-2024018564-A1
Application number	US-202118251167-A
Country	US
Kind code	A1
Filing date	Nov 1, 2021
Priority date	Oct 30, 2020
Publication date	Jan 18, 2024
Grant date	—

Abstract

The present invention is related to a fermentative production of retinoids, including retinol or retinyl acetate, comprising cultivation of a retinoid-producing host cell, such as fungal host cell, particularly oleaginous host such as e.g. Yarrowia , in a two-phase culture system in the presence of vegetable oil as second phase. The retinoids present in the vegetable oil can be used without further isolation or purification steps as active ingredients in the food, feed, pharma or cosmetic industry.

First claim

1 . Process for in situ extraction of retinoids in a fermentation process, comprising cultivation of the host cell, particularly fungal host cell, preferably oleaginous yeast, more preferably Yarrowia , in the presence of a second phase lipophilic solvent comprising vegetable oil.

2 . Process according to claim 1 , wherein the host cell is grown on a carbon source selected from glucose or ethanol.

3 . Process according to claim 1 , wherein the percentage of retinoids is increased by at least about 10% compared to a process using silicone oil as second phase lipophilic solvent.

4 . Process according to claim 1 , wherein the solvent is used for collection and extraction of the fermentative produced retinoids.

5 . Process according to claim 1 , wherein the fermentative produced retinoids comprise a percentage of at least about 0.001 to about 30% retinyl acetate based on total retinoids.

6 . Process according to claim 1 , wherein the vegetable oil is selected from the group consisting of oleic, palmitic, steric or linoleic acid and glycerol, preferably selected from the group consisting of corn oil, olive oil, cottonseed oil, rapeseed oil, sesame oil, canola oil, safflower oil, sunflower oil, soybean oil, grapeseed oil, and peanut oil, more preferably selected from corn oil.

7 . Process according to claim 1 , wherein the host cell is selected from oleaginous host cell including Rhodosporidium, Lipomyces or Yarrowia , preferably Yarrowia lipolytica.

8 . Process according to claim 1 , wherein the host cell comprises one or more genetic modifications, particularly reduction or abolishment of endogenous lipase activity, preferably reduction or abolishment of lipase activity corresponding to Yarrowia lipase activity 2, 3, 4, and/or 8, more preferably Yarrowia lipase activity 8.

9 . Process according to claim 1 , wherein the host cell is transformed with and is expressing heterologous genes, preferably genes encoding acetylating enzymes catalyzing the conversion of retinol to retinyl acetate, more preferably fungal enzymes, most preferably originated from Lachancea mirantina.

10 . Process according to claim 9 , wherein the acetylating enzyme comprises one or more amino acid substitution(s) in a sequence with at least about 20%, such as e.g. 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 92, 95, 97, 98, 99% or up to 100% identity to SEQ ID NO:12, wherein the one or more amino acid substitution(s) are located at position(s) corresponding to amino acid residue(s) selected from the group consisting of position 68, 69, 72, 73, 171, 174, 176, 178, 291, 292, 294, 301, 307, 308, 311, 312, 320, 322, 334, 362, 405, 407, 409, 480, 483, 484, 490, 492, 520, 521, 522, 524, 525, 526 and combinations thereof in a polypeptide according to SEQ ID NO:12.

Assignees

• Dsm Ip Assets Bv

Inventors

• Houston Peter Louis
• Vyas Valmik Kanubhai

Classifications

• C12R2001/645

  Fungi {; Processes using fungi} · CPC title

• C12N1/063

  of yeast · CPC title

• C12P23/00 Primary

  Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes (containing heterorings C12P17/00) · CPC title

• C12N9/20

  Triglyceride splitting, e.g. by means of lipase · CPC title

• C12Y301/01003

  Triacylglycerol lipase (3.1.1.3) · CPC title