Process for the production of gamma-aminobutyric acid
AssigneeBASF SE
InventorsOskar Zelder, Weol Kyu Jeong, Corinna Klopprogge, Andrea Herold, Hartwig Schröder
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Abstract
The present invention relates to a novel method for the fermentative production of gamma-aminobutyric acid (GABA) by cultivating a recombinant microorganism expressing an enzyme having a glutamate decarboxylase activity. The present invention also relates to corresponding recombinant hosts, recombinant vectors, expression cassettes and nucleic acids suitable for preparing such hosts as well as to a method for preparing polyamides making use of GABA as obtained fermentative production.
Claims21
A method for the fermentative production of gamma-aminobutyric acid (GABA), comprising cultivating a recombinant microorganism derived from a parent microorganism having the ability to produce glutamate and additionally having the ability to express a heterologous glutamate decarboxylase (E.C. 4.1.1.15) in a culture medium, wherein said microorganism is a Corynebacterium,wherein glutamate is converted to GABA in said recombinant microorganism, and wherein said heterologous glutamate decarboxylase is encoded by a nucleic acid composing:
- a) the nucleotide sequence from position 472 to 1200 of SEQ ID NO:
- 1 or from position 193 to 1605 of SEQ ID NO:
- 1;
- b) a nucleotide sequence encoding a glutamate decarboxylase comprising the amino acid sequence of SEQ ID NO:
- 2;
- or
- c) a nucleotide sequence encoding a glutamate decarboxylase comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 2, wherein said glutamate decarboxylase comprises the amino acid sequence from Thr94 to Leu336 of SEQ ID NO:
- 2 or an amino acid sequence having at least 98% sequence identity to the amino acid sequence from Thr94 to Leu336 of SEQ ID NO:
- 2.
A chimeric glutamate decarboxylase comprising the amino acid sequence from Thr94 to Leu336 of SEQ ID NO:
- 2 or an amino acid sequence having at least 95% sequence identity thereto, wherein said chimeric glutamate decarboxylase further comprises at least one amino acid sequence portion at the N-terminus, the C-terminus, or both the N-terminus and C-terminus, of said amino acid sequence from Thr94 to Leu336 of SEQ ID NO:
- 2 or an amino acid sequence having at least 95% sequence identity thereto, wherein the at least one amino acid sequence portion is obtained from a corresponding amino acid sequence portion of a glutamate decarboxylase of a plant different from Solanum tuberosum, and wherein said chimeric glutamate decarboxylase encoded by a nucleic acid comprising:
- a) the nucleotide sequence from position 472 to 1200 of SEQ ID NO:
- 1 or from position 193 to 1605 of SEQ ID NO:
- 1;
- b) a nucleotide sequence encoding a glutamate decarboxylase comprising the amino acid sequence of SEQ ID NO:
- 2;
- or
- c) a nucleotide sequence encoding a glutamate decarboxylase comprising an amino acid sequence having at least 95% sequence identity to SEQ ID NO:
- 2, wherein said glutamate decarboxylase comprises the amino acid sequence from Thr94 to Leu336 of SEQ ID NO:
- 2 or an amino acid sequence having at least 98% sequence identity to the amino acid sequence from Thr94 to Leu336 of SEQ ID NO:
- 2.
A method for the fermentative production of gamma-aminobutyric acid (GABA), comprising cultivating a recombinant microorganism derived from a parent microorganism having the ability to produce glutamate and additionally having the ability to express a heterologous glutamate decarboxylase (E.C. 4.1.1.15) in a culture medium, wherein said microorganism is a Corynebacterium, wherein glutamate is converted to GABA in said recombinant microorganism, and wherein said heterologous glutamate decarboxylase is encoded by a nucleotide sequence which hybridizes to the complementary sequence of the nucleotide sequence from position 472 to 1200 of SEQ ID NO:
- 1 or from position 193 to 1605 of SEQ ID NO:
- 1 under stringent hybridization conditions comprising hybridization at 42°C in a solution consisting of 50% formamide, 5×SSC, 50mM sodium phosphate, 5×Denhardt Solution, 10% dextran sulfate and 20g/ml denatured, sheared salmon sperm DNA, followed by washing with 0.1×SSC at 65°C.
Dependencies are inferred from the claim text.