Method for production of recombinant Erwinia asparaginase
AssigneePfenex Inc.
InventorsRussell J. Coleman, Torben Bruck
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Abstract
Provided herein are methods of production of recombinant Erwinia asparaginase. Methods herein produce asparaginase having high expression levels in the periplasm or the cytoplasm of the host cell having activity comparable to commercially available asparaginase preparations.
Claims37
A method for producing a recombinant type II asparaginase, the method comprising culturing a Pseudomonadales host cell in a culture medium and expressing the recombinant type II asparaginase in the cytoplasm of the Pseudomonadales host cell from an expression construct comprising a nucleic acid encoding the recombinant type II asparaginase, wherein the recombinant type II asparaginase encoded by the nucleic acid comprises an amino acid sequence at least about 85% identical to SEQ ID NO:
- 1, and wherein the recombinant type II asparaginase is produced in the cytoplasm at a yield of about 20% to about 50% total cell protein (TCP) in soluble form.
A method for producing a recombinant type II asparaginase, the method comprising:
- culturing a Pseudomonadales host cell in a culture medium and expressing the recombinant type II asparaginase in the periplasm of the host cell from an expression construct comprising a nucleic acid encoding the recombinant type II asparaginase, wherein the recombinant type II asparaginase encoded by the nucleic acid comprises an amino acid sequence at least about 85% identical to SEQ ID NO:
- 1, and wherein the recombinant type II asparaginase is produced in the periplasm at a yield of about 20% to about 40% TCP in soluble form.
Dependencies are inferred from the claim text.