Recombinant host cell with altered membrane lipid composition

Patent metadata

Field	Value
Publication number	US-11479798-B2
Application number	US-201816499072-A
Country	US
Kind code	B2
Filing date	Mar 28, 2018
Priority date	Mar 29, 2017
Publication date	Oct 25, 2022
Grant date	Oct 25, 2022

Abstract

The present invention is in the field of recombinant biotechnology, in particular in the field of protein expression. The invention generally relates to a method of expressing a protein of interest (POI) from a host cell. The invention relates particularly to improving a host cell's capacity to express and/or secrete a protein of interest and use of the host cell for protein expression. The invention also relates to cell culture technology, and more specifically to culturing cells to produce desired molecules for medical purposes or food products.

First claim

The invention claimed is:

1. A method of increasing the yield of a protein of interest in a eukaryotic host cell, comprising overexpressing in said host cell at least one polynucleotide encoding at least one protein which is involved in lipid metabolism, thereby increasing the yield of said protein of interest in comparison to a host cell which does not overexpress a polynucleotide encoding a protein which is involved in lipid metabolism,
wherein said protein which is involved in lipid metabolism is involved in sphingolipid biosynthesis and comprises an amino acid sequence as shown in SEQ ID NO: 1 or a homologue thereof having at least 70% sequence identity to an amino acid sequence as shown in SEQ ID NO: 1.

2. A method of manufacturing a protein of interest according to claim 1 comprising:
providing the host cell engineered to overexpress at least one polynucleotide encoding at least one protein, wherein said protein which is involved in lipid metabolism is involved in sphingolipid biosynthesis and comprises an amino acid sequence as shown in SEQ ID NO: 1 or a homologue thereof, wherein the homologue has at least 70% sequence identity to an amino acid sequence as shown in SEQ ID NO: 1, wherein said host cell comprises a heterologous polynucleotide encoding a protein of interest;
culturing the host cell under suitable conditions to overexpress the protein involved in lipid metabolism or homologue thereof and to express said protein of interest; and optionally
isolating said protein of interest from the cell culture.

3. A recombinant eukaryotic host cell for manufacturing a protein of interest, wherein the host cell is engineered to overexpress at least one polynucleotide encoding at least one protein which is involved in lipid metabolism, wherein said protein which is involved in lipid metabolism is involved in sphingolipid biosynthesis and comprises an amino acid sequence as shown in SEQ ID NO: 1 or a homologue thereof, wherein the homologue has at least 70% sequence identity to an amino acid sequence as shown in SEQ ID NO: 1.

4. The method of claim 1 , wherein the host cell is selected from the group consisting of Pichia pastoris, Hansenula polymorpha, Trichoderma reesei, Saccharomyces cerevisiae, Kluyveromyces lactis, Yarrowia lipolytica, Pichia methanolica, Candida boidinii, Komagataella sp., Aspergillus sp. and Schizosaccharomyces pombe.

5. The method of claim 2 , wherein the host cell is selected from the group consisting of Pichia pastoris, Hansenula polymorpha, Trichoderma reesei, Saccharomyces cerevisiae, Kluyveromyces lactis, Yarrowia lipolytica, Pichia methanolica, Candida boidinii, Komagataella sp., Aspergillus sp. and Schizosaccharomyces pombe.

6. The recombinant eukaryotic host cell of claim 3 , wherein the host cell is selected from the group consisting of Pichia pastoris, Hansenula polymorpha, Trichoderma reesei, Saccharomyces cerevisiae, Kluyveromyces lactis, Yarrowia lipolytica, Pichia methanolica, Candida boidinii, Komagataella sp., Aspergillus sp. and Schizosaccharomyces pombe.

Assignees

• Boehringer Ingelheim Rcv Gmbh
• Validogen Gmbh
• Lonza Ag
• Sandoz Ag
• Biomin Holding Gmbh

Inventors

• Grillitsch Karlheinz
• Daum Guenther
• Grutsch Andreas

Classifications

• C12P21/02 Primary

  having a known sequence of two or more amino acids, e.g. glutathione · CPC title

• C12N15/67 Primary

  General methods for enhancing the expression · CPC title

• C12N15/815

  for yeasts other than Saccharomyces · CPC title

• C12N15/63 Primary

  Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression · CPC title

• C07K16/00

  Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies · CPC title