Methods and compositions for producing proteins of interest in pigment deficient bacillus cells
AssigneeDanisco US Inc.
InventorsRyan L. Frisch, Ibrahim Halloum, Mikhail Karymov, Christopher John Webb
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Abstract
Certain embodiments of the disclosure are related to compositions and methods for producing proteins of interest in pigment deficient Bacillus cells. Certain other embodiments are related to compositions and methods for obtaining pigment deficient Bacillus cells. Certain other embodiments are related to compositions and methods for growing/cultivating/fermenting pigment deficient Bacillus cells. Certain other embodiments are therefore related to compositions and methods for producing, isolating, recovering and the like proteins of interest that are pigment deficient.
Claims14
A recombinant Bacillus cell obtained by genetically modifying a parental Bacillus cell, wherein the recombinant cell comprises an introduced yvmA expression cassette comprising a heterologous promoter operably linked to a yvmA open reading frame (ORF) encoding a functional YvmA protein having at least 95% identity to SEQ ID NO:
- 30, wherein the recombinant cell produces at least 5% less pulcherrimin than the parental cell when fermented under the same conditions, as determined by (i) pelleting an aliquot of fermentation culture, (ii) washing the pellet with methanol and water, (iii) extracting the pellet with 2N NaOH, and (iv) measuring absorbance at about 405-410 nm.
A method for cultivating a Bacillus cell deficient in the production of a red pigment comprising:
- modifying a parental Bacillus cell by introducing into the parental cell a yvmA expression cassette comprising a heterologous promoter operably linked to a yvmA open reading frame (ORF) encoding a functional YvmA protein having at least 95% identity to SEQ ID NO:
- 30, and
- cultivating the recombinant cell under suitable conditions,
- wherein the recombinant cell produces at least 5% less pulcherrimin than the parental cell when cultivated under the same conditions, as determined by (i) pelleting an aliquot of culture, (ii) washing the pellet with methanol and water, (iii) extracting the pellet with 2N NaOH, and (iv) measuring absorbance at about 405-410 nm.
A method for producing a heterologous protein of interest (POI) comprising:
- modifying a parental Bacillus cell expressing a heterologous POI by introducing into the parental cell an expression cassette comprising a heterologous promoter operably linked to a yvmA open reading frame (ORF) encoding a functional YvmA protein having at least 95% identity to SEQ ID NO:
- 30, and
- fermenting the recombinant cell under suitable conditions for the production of the POI,
- wherein the recombinant cell produces at least 5% less pulcherrimin than the parental cell when fermented under the same conditions, as determined by (i) pelleting an aliquot of culture, (ii) washing the pellet with methanol and water, (iii) extracting the pellet with 2N NaOH, and (iv) measuring absorbance at about 405-410 nm.
Dependencies are inferred from the claim text.