Inducible cell lysis system
AssigneeMotif FoodWorks, Inc.
InventorsLennart BIERMANN, Marius HENKEL, Lars LILGE, Rudolf HAUSMANN, Philippe PROCHASSON
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Abstract
A programmable cell-lysis system and related compositions and methods are described, which facilitate expression and harvest of recombinant proteins from host cells. The programmable cell-lysis system comprises a host cell comprising a polynucleotide sequence encoding a cell lysis promoting polypeptide or peptide operably linked to an inducible promoter. The cell lysis systems can be configured to produce various heterologous polypeptides for different applications.
Claims40
An engineered host cell, wherein the host cell comprises:
- a. modification of at least one nucleic acid sequence encoding a sporulation-promoting polypeptide, a cell lysis inhibitor polypeptide or a combination thereof;
- and optionally,
- b. a first polynucleotide sequence encoding at least one cell lysis promoting polypeptide.
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A method of producing a heterologous polypeptide, the method comprising:
- a. culturing an engineered host cell, wherein the engineered host cell comprises:
- i. modification of at least one nucleic acid sequence encoding a sporulation-promoting polypeptide, a cell lysis inhibitor polypeptide or a combination thereof;
- ii. an optional first polynucleotide sequence encoding at least one cell lysis promoting polypeptide operably linked to a first promoter;
- and
- iii. a second polynucleotide sequence comprising a second nucleic acid sequence encoding a heterologous polypeptide operably linked to a second promoter;
- b. expressing the heterologous polypeptide in the host cell;
- c. inducing cell lysis by maintaining the host cell for a time and under conditions sufficient for expression of at least one cell lysis promoting polypeptide;
- d. harvesting the heterologous polypeptide from culture supernatant.
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An engineered cell lysis system for production of a heterologous polypeptide, the system comprising an engineered host cell, wherein the engineered host cell comprises:
- i. modification of at least one nucleic acid sequence encoding a sporulation-promoting polypeptide, a cell lysis inhibitor polypeptide or a combination thereof;
- ii. an optional first polynucleotide sequence encoding at least one cell lysis promoting polypeptide operably linked to a first promoter;
- and
- iii. a second polynucleotide sequence comprising a second nucleic acid sequence encoding a heterologous polypeptide operably linked to a second promoter.
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Dependencies are inferred from the claim text.