Methods of increasing the production of cobalamins using cob gene expression
AssigneeRhone-Poulenc Biochimie, et al.
InventorsFrancis Blanche, Beatrice Cameron, Joel Crouzet, Laurent Debussche, Sophie Levy Schil, Denis Thibaut
Life of this patent
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Abstract
Novel polypeptides involved in the biosynthesis of cobalamines and/or cobamides, in particular coenzyme B12, genetic material responsible for expressing these polypeptides, and a method for preparing them, are described. A method for amplifying the production of cobalamines, and particularly coenzyme B12, using recombinant DNA techniques, are also described.
Claims58
A method for increasing the production of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein said method comprises:
- a) introducing a plasmid comprising a DNA sequence selected from the group consisting of the cobA, cobB, cobC, cobD, cobE, cobF, cobG, cobH, cobI, cobJ, cobK, cobL, cobM, cobN, cobO, cobP, cobQ, cobS, cobT, cobU, cobV, cobW, and cobX genes of P. denitrificans and homologs of said genes resulting from the degeneracy of the genetic code into a microorganism capable of producing cobalamins or cobamides;
- b) culturing said microorganism under conditions suitable for the synthesis of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein said culture conditions are also suitable for expression of said DNA;
- and
- c) recovering the cobalamins, cobamides, cobalamin precursors, or cobamide precursors produced.
A method for increasing the production of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein said method comprises:
- a) introducing a DNA selected from the group consisting of the cobA, cobB, cobC, cobD, cobE, cobF, cobG, cobH, cobI, cobJ, cobK, cobL, cobM, cobN, cobO, cobP, cobQ, cobS, cobT, cobU, cobV, cobW, and cobX genes of P. denitrificans and homologs of said genes resulting from the degeneracy of the genetic code into a microorganism capable of producing cobalamins or cobamides;
- b) culturing said microorganism under conditions suitable for the synthesis of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein said culture conditions are also suitable for expression of said DNA;
- and
- c) recovering the cobalamins, cobamides, cobalamin precursors, or cobamide precursors produced.
A method for increasing the industrial production of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein said method comprises:
- a) introducing at least one plasmid comprising a DNA sequence selected from the group consisting of cobA, cobB, cobC, cobD, cobE, cobF, cobG, cobH, cobI, cobJ, cobK, cobL, cobM, cobN, cobO, cobP, cobQ, cobS, cobT, cobU, cobV, cobW, and cobX genes of P. denitrificans and homologs of said genes resulting from the degeneracy of the genetic code into a microorganism producing cobalamins or cobamides;
- b) culturing said microorganism under conditions suitable for the synthesis of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein industrial production comprises culture conditions suitable for expression of said DNA and suitable for production of at least 100 grams of cells;
- and
- c) recovering the cobalamins, cobamides, cobalamin precursors, or cobamide precursors produced,
- wherein the industrial production of cobalamins, cobamides, cobalamin precursors, or cobamide precursors by said microorganism is increased by the introduction of said plasmid.
A method for increasing the industrial production of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein said method comprises:
- a) introducing at least one plasmid comprising a DNA sequence selected from the group consisting of cobA, cobB, cobC, cobD, cobE, cobF, cobG, cobH, cobI, cobJ, cobK, cobL, cobM, cobN, cobO, cobP, cobQ, cobS, cobT, cobU, cobV, cobW, and cobX genes of P. denitrificans and homologs of said genes resulting from the degeneracy of the genetic code into a microorganism capable of producing cobalamins or cobamides;
- b) culturing said microorganism under conditions suitable for the synthesis of cobalamins, cobamides, cobalamin precursors, or cobamide precursors, wherein industrial production comprises culture conditions suitable for expression of said DNA and suitable for production of at least 100 grams of cells;
- and
- c) recovering the cobalamins, cobamides, cobalamin precursors, or cobamide precursors produced,
- wherein the industrial production of cobalamins, cobamides, cobalamin precursors, or cobamide precursors by said microorganism is increased by the introduction of said plasmid.
The method of any one of claims 45-49, wherein said cobalamin is coenzyme B12.
Dependencies are inferred from the claim text.