Method and agents for producing N-acetylneuraminic acid (NeuNAc)
AssigneeTechnische UniversitäT Wien
InventorsAstrid Mach-Aigner, Robert Mach, Matthias G. Steiger
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Abstract
The invention relates to an isolated nucleic acid molecule comprising at least one promoter that is active in fungal cells of the trichoderma species, wherein a nucleic acid sequence encoding an N-acetylglucosamine-2-epimerase and/or an N-acetylneuraminic acid synthase is operatively bound to each promoter. The at least one promoter that is active in fungal cells is a constitutive promoter.
Claims18
A method for producing N-acetylneuraminic acid (NeuNAc), comprising cultivating a fungal cell in the presence of an N-acetyl-D-glucosamine source,
- wherein the fungal cell is of the genus Trichoderma and comprises a) a nucleic acid molecule or b) a set of two nucleic acid molecules,
- wherein the nucleic acid molecule of a) and each of the nucleic acid molecules of the set b) comprise at least one constitutive promoter that is active in the genus Trichoderma, and wherein the nucleic acid molecule of a) further comprises a nucleic acid sequence encoding an N-acetylglucosamine 2-epimerase and an NeuNAc synthase operatively linked to one or more of said at least one promoter, or the set of two nucleic acid molecules of b) comprises a nucleic acid molecule encoding the N-acetylglucosamine 2-epimerase and a nucleic acid molecule encoding the NeuNAc synthase, each of said molecules comprising at least one promoter that is active in fungal cells of the genus Trichoderma operably linked to the sequence encoding N-acetylglucosamine 2-epimerase or N-acetylneuraminic acid synthase,
- wherein the N-acetylglucosamine 2-epimerase is SEQ ID NO:1.
A method for producing N-acetylneuraminic acid (NeuNAc), comprising cultivating a fungal cell in the presence of an N-acetyl-D-glucosamine source,
- wherein the fungal cell is of the genus Trichoderma and comprises a) a nucleic acid molecule or b) a set of two nucleic acid molecules,
- wherein the nucleic acid molecule of a) and each of the nucleic acid molecules of the set b) comprise at least one constitutive promoter that is active in the genus Trichoderma, and wherein the nucleic acid molecule of a) further comprises a nucleic acid sequence encoding an N-acetylglucosamine 2-epimerase and an NeuNAc synthase operatively linked to one or more of said at least one promoter, or the set of two nucleic acid molecules of b) comprises a nucleic acid molecule encoding the N-acetylglucosamine 2-epimerase and a nucleic acid molecule encoding the NeuNAc synthase, each of said molecules comprising at least one promoter that is active in fungal cells of the genus Trichoderma operably linked to the sequence encoding N-acetylglucosamine 2-epimerase or N-acetylneuraminic acid synthase,
- wherein the NeuNAc synthase is SEQ ID NO:2.
Dependencies are inferred from the claim text.