Compositions and methods for improved protein production
AssigneeDanisco US Inc.
InventorSusan M. MADRID
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Abstract
Aspects of the present disclosure are drawn to methods of improving the expression of secreted cuproenzymes from host cells by manipulating the expression level of one or more proteins involved in copper transport in the host cell, e.g., membrane-bound copper transporting ATPases and soluble copper transporters. The present disclosure also provides compositions containing such improved host cells as well as products derived from the improved host cells that contain one or more cuproenzymes of interest.
Claims29
A method for producing a cuproenzyme from a host cell comprising:
- overexpressing a copper metallochaperone in a host cell that expresses a cuproenzyme, and culturing the host cell under conditions sufficient to produce the cuproenzyme, wherein the host cell produces an increased amount of the cuproenzyme as compared to a corresponding host cell that does not overexpress the copper metallochaperone when cultured under substantially the same culture conditions.
11-16. (canceled)
20-24. (canceled)
A recombinant host cell comprising:
- a first polynucleotide encoding a cuproenzyme, and a second polynucleotide encoding a copper metallochaperone, wherein the cuproenzyme is expressed in the host cell and the copper metallochaperone is over-expressed in the host cell, and wherein the level of expression of the cuproenzyme is increased in the host cell as compared to a corresponding host cell that does not overexpress the copper metallochaperone under substantially the same culture conditions.
(canceled)
29-30. (canceled)
The recombinant host cell of claim 30, wherein host cell is T reesei and the promoter is a pki or a cbh1 promoter derived from T reesei.
33-35. (canceled)
(canceled)
Dependencies are inferred from the claim text.