Preparation of an esterase
View Patent ↗The present invention relates to a recombinant E. coli strain characterized in that the organisms contains a gene coding for a protein having esterase activity. Such E. coli is suitable for the preparation of a protein with esterase activity, whereby the expression takes place without coexpression of GroEL and/or GroES from a plasmid. The expression of glutathione reductase and/or thioredoxin reductase may be abolished by mutation. Furthermore, the ability of the organism to grow under oxygen-rich conditions has been restored by mutation. The expression of the gene of the protein with esterase activity is unaccompanied by the expression of an additional gene encoding a heat shock chaperone protein.
1. A method of producing a protein with esterase activity comprising expressing a gene encoding the protein in an E. coli strain, wherein the gene encoding the protein with esterase activity has at least 99% identity to the polynucleotide of SEQ ID NO 11 and encodes a protein that has at least 99% identity to SEQ ID NO 12 and wherein the expression takes place without coexpression of GroEL and/or GroES from a plasmid.
2. The method according to claim 1 , wherein the expression takes place without coexpression of an additional heat shock chaperone protein.
3. The method according to claim 1 , wherein the expression takes place with overexpression of endogenous dsbC gene.
4. The method according to claim 1 , wherein the E. coli strain comprises Origami 1, Origami 2 or Origami B cells.
5. The method according to claim 4 , wherein the expression takes place without coexpression of an additional heat shock chaperone protein.
6. The method according to claim 4 , wherein the expression takes place with overexpression of endogenous dsbC gene.
7. The method according to claim 1 , wherein the expression takes place in a recombinant E. coli strain that is unable to express glutathione reductase and/or thioredoxin reductase.
8. The method according to claim 7 , wherein the expression takes place without coexpression of an additional heat shock chaperone protein.
9. The method according to claim 7 , wherein the expression takes place with overexpression of endogenous dsbC gene.
10. A method of producing a protein with esterase activity comprising expressing a gene encoding the protein in an E. coli strain, wherein the gene encoding the protein with esterase activity has at least 99% identity to the polynucleotide of SEQ ID NO 11 and encodes a protein that has at least 99% identity to SEQ ID NO 12.
11. The method according to claim 10 , wherein the expression takes place without coexpression of an additional heat shock chaperone protein.
12. The method according to claim 10 , wherein the expression takes place with overexpression of endogenous dsbC gene.
13. The method according to claim 10 , wherein the E. coli strain comprises Origami 1, Origami 2 or Origami B cells.
14. The method according to claim 13 , wherein the expression takes place without coexpression of an additional heat shock chaperone protein.
15. The method according to claim 13 , wherein the expression takes place with overexpression of endogenous dsbC gene.
16. The method according to claim 10 , wherein the expression takes place in a recombinant E. coli strain that is unable to express glutathione reductase and/or thioredoxin reductase.
17. The method according to claim 16 , wherein the expression takes place without coexpression of an additional heat shock chaperone protein.
18. The method according to claim 16 , wherein the expression takes place with overexpression of endogenous dsbC gene.