Method for the fermentative production of D-pantothenic acid using Coryneform bacteria
AssigneesDegussa-Huls AG, Forschunszentrum Julich GmbH
InventorsLothar Eggeling, Georg Thierbach, Hermann Sahm
Life of this patent
Estimating the term…
Abstract
The invention discloses three polynucleotide sequences for the fermentative production of D-pantothenic acid. These polynucleotide sequences are genes named panB, encoding a ketopantoate hydroxymethyltransferase, panC, encoding pantothenate synthase, and ilvD, encoding dihydroxy-acid dehydratase. The genes panB and panC are found on the same operon, panBC, while the gene ilvD is found in a separate operon. These genes can be used separately or together to enhance the production of D-pantothenic acid in microorganisms, especially in Corynebacterium.
Claims25
An isolated polynucleotide from Corynebacterium comprising a nucleotide sequence selected from the group consisting of:
- a) a nucleotide sequence encoding the panB gene product, which is ketopantoate hydroxymethyltransferase, whose amino acid sequence is set forth in SEQ ID NO:
- 3;
- and
- b) a nucleotide sequence encoding the panC gene product, which is pantothenate synthetase, whose amino acid sequence is set forth in SEQ ID NO:
- 4.
An isolated polynucleotide from Corynebacterium comprising a nucleotide sequence which hybridizes to the antisense strand of SEQ ID NO:
- 1 and that encodes an enzyme selected from the group consisting of ketopantoate hydroxymethyltransferase and pantothenate synthetase.
A method for producing pantothenic acid comprising:
- a) transforming a vector into a microorganism to produce a recombinant microorganism, wherein said vector comprises a panB gene and a panC gene, both genes being operably linked to a suitable regulatory sequence;
- b) growing said recombinant microorganism under conditions suitable for the production of pantothenic acid in an appropriate culture medium;
- and
- c) recovering pantothenic acid from said culture medium;
- wherein said panB gene encodes ketopantoate hydroxymethyltransferase, said panC gene encodes pantothenate synthetase, and both said panB and said panC genes originate from Corynebacterium.
A method for producing pantothenic acid in Corynebacterium comprising:
- a) inserting an expression cassette upstream from the panB and panC genes in said Corynebacterium;
- b) growing said Corynebacterium under conditions suitable for the production of pantothenic acid in an appropriate culture medium;
- and
- c) recovering pantothenic acid from said culture medium.
A method for producing pantothenic acid in Corynebacterium comprising:
- a) inserting expression cassettes upstream from the panB, panC and ilvD genes in said Corynebacterium;
- b) growing said Corynebacterium under conditions suitable for the production of pantothenic acid in an appropriate culture medium;
- and
- c) recovering pantothenic acid from said culture medium.
A method for producing pantothenic acid in Corynebacterium comprising:
- a) increasing the stability of the mRNA which is translated from the panB and panC genes in said Corynebacterium;
- and/or
- b) preventing the degradation of the panB and panC gene products in said Corynebacterium, which gene products are ketopantoate hydroxymethyltransferase and pantothenate synthetase, respectively;
- wherein said Corynebacterium is grown under conditions suitable for the production of pantothenic acid in an appropriate culture medium, and wherein pantothenic acid is recovered from said culture medium.
Dependencies are inferred from the claim text.