Production of heterologous extracellular polypeptides in Yarrowia lipolytica
AssigneeCsir
InventorsPetrus Jackobus Van Zyl, Mulalo Bethuel Nthangeni, Faranani Ramagoma
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Abstract
This invention relates to protein and/or polypeptide production, particularly improved production of extracellular heterologous polypeptides and proteins. In particular, the invention relates to compositions of cell populations capable of improved levels of extracellular secretion relative to control populations, kits containing such compositions, methods of producing heterologous proteins of interest and recombinant microorganisms capable of improved extracellular heterologous protein production.
Claims12
A composition comprising a Yarrowia lipolytica-derived cell population (Accession No. NCIMB 42022, deposited on 9 Aug. 2012 at National Collection of Industrial Food and Marine Bacteria (NCIMB Ltd), of Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland) capable of expressing a heterologous polypeptide of interest, wherein the level of extracellular secretion of the heterologous polypeptide is increased relative to the level of extracellular secretion of the heterologous polypeptide by a control Yarrowia lipolytica cell population (Accession No. NCIMB 42021, deposited on 9 Aug. 2012 at National Collection of Industrial Food and Marine Bacteria (NCIMB Ltd), of Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland), wherein the Yarrowia lipolytica-derived cell population has at least one genomic modification affecting one or more genes involved in glycoslyphosphatidylinositol (GPI) anchoring relative to the control Yarrowia lipolytica cell population, wherein the genomic modification is attributable to the increase in extracellular secretion.
A method of producing a heterologous polypeptide of interest comprising:
- a. obtaining a Yarrowia lipolytica-derived cell population (Accession No. NCIMB 42022, deposited on 9 Aug. 2012 at National Collection of Industrial Food and Marine Bacteria (NCIMB Ltd), of Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland) comprising an expression cassette encoding the heterologous polypeptide of interest;
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- b. culturing the cell population under conditions sufficient for expression of the heterologous polypeptide of interest, wherein the Yarrowia lipolytica-derived cell population has at least one genomic modification affecting one or more genes involved in glycoslyphosphatidylinositol (GPI) anchoring relative to a control Yarrowia lipolytica cell population (Accession No. NCIMB 42021, deposited on 9 Aug. 2012 at National Collection of Industrial Food and Marine Bacteria (NCIMB Ltd), of Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA, Scotland), wherein the genomic modification results in an increase in extracellular secretion of the heterologous polypeptide of interest.
A method for increasing the extracellular secretion of polypeptides in Yarrowia lipolytica strains wherein the increased extracellular secretion of the polypeptide of interest into a culture medium through the cell membrane is triggered by affecting one or more genes involved in glycosylphosphatidylinositol (GPI) anchoring.
A recombinant Yarrowia lipolytica microorganism genomically modified such that one or more genes involved in glycosylphosphatidylinositol (GPI) anchoring are affected, the microorganism transformed with a recombinant vector comprising a target polypeptide-encoding gene and capable of expressing the target polypeptide for secretion extracellularly, the recombinant microorganism further having a characteristic that at least one gene involved in glycosylphosphatidylinositol (GPI) anchoring is affected and the target polypeptide-encoding gene operates to express the target polypeptide such that the target polypeptide is secreted extracellularly of the microorganism.
Dependencies are inferred from the claim text.