Methods for reducing norleucine misincorporation into proteins using a microorganism comprising a mutant metA allele
AssigneeGenentech, Inc.
InventorsMichael W. Laird, Karthik Veeravalli
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Abstract
The present invention relates to methods and compositions for preventing incorporation of norleucine into proteins during recombinant protein production in bacteria. The present invention also provides microorganism host cells and nucleic acid molecules for use with the methods and compositions provided herein.
Claims29
A method for reducing norleucine misincorporation into a protein or a polypeptide, the method comprising expressing the protein or the polypeptide in a microorganism, wherein the microorganism comprises a mutant metA allele, wherein the mutant metA allele comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26.
A method for reducing norleucine misincorporation into a protein or a polypeptide, the method comprising expressing the protein or the polypeptide in a microorganism, wherein the microorganism comprises a mutant metK allele, wherein the mutant metK allele comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:27 and SEQ ID NO:28.
A method for reducing norleucine misincorporation into a protein or a polypeptide, the method comprising expressing the protein or the polypeptide in a microorganism, wherein the microorganism comprises a mutant metA allele and a mutant metK allele, wherein the mutant metA allele comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26, and further wherein the mutant metK allele comprises a nucleic acid sequence selected from the group consisting of SEQ ID NO:27and SEQ ID NO:28.
A method for producing a protein or a polypeptide in an E. coli host cell, wherein the protein or the polypeptide contains no detectable levels of norleucine residues, the method comprising expressing in the E. coli host cell a nucleic acid encoding the protein or the polypeptide under culture conditions suitable to allow for expression of the protein or the polypeptide, wherein the E. coli host cell comprises a mutant metA allele, wherein the mutant metA allele encodes for MetA having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, and SEQ ID NO:26.
A method for producing a protein or a polypeptide in an E. coli host cell, wherein the protein or the polypeptide contains no detectable levels of norleucine residues, the method comprising expressing in the E. coli host cell a nucleic acid encoding the protein or the polypeptide under culture conditions suitable to allow for expression of the protein or the polypeptide, wherein the E. coli host cell comprises a mutant metK allele, wherein the mutant metK allele encodes for MetK having an amino acid sequence encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO:27 and SEQ ID NO:28.
Dependencies are inferred from the claim text.