Method and agents for producing n-acetylneuraminic acid (neunac)

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.

1 . An isolated nucleic acid molecule comprising at least one promoter that is active in fungal cells of the genus  Trichoderma  and has a nucleic acid sequence encoding an N-acetylglucosamine 2-epimerase and/or an N-acetylneuraminic acid synthase operably linked thereto, wherein said at least one promoter that is active in fungal cells is a constitutive promoter. 
     
     
         2 . The isolated nucleic acid molecule according to  claim 1 , characterized in that the fungal cells of the genus  Trichoderma  are  Trichoderma reesei  cells. 
     
     
         3 . The isolated nucleic acid molecule according to  claim 1 , characterized in that the constitutive promoter is selected from the group consisting of promoters of the glycolytic genes, especially of pki, gpd or zwf1, tef1a, act, cox4, neg1 and sar1. 
     
     
         4 . A vector comprising the nucleic acid molecule according to  claim 1 . 
     
     
         5 . A fungal cell of the genus  Trichoderma  comprising the nucleic acid molecule according to  claim 1 . 
     
     
         6 . The fungal cell according to  claim 5 , characterized in that the fungal cell of the genus  Trichoderma  is a  Trichoderma reesei  cell. 
     
     
         7 . The fungal cell according to  claim 5 , characterized in that it comprises at least one nucleic acid molecule whose nucleic acid sequence encodes an N-acetylglucosamine 2-epimerase and an N-acetylneuraminic acid synthase and is operably linked to a constitutive promoter that is active in fungal cells. 
     
     
         8 . A method for producing N-acetylneuraminic acid (NeuNAc) comprising cultivating fungal cells of the genus  Trichoderma  according to  claim 5  in the presence of an N-acetyl-D-glucosamine source. 
     
     
         9 . The method according to  claim 8 , characterized in that the N-acetyl-D-glucosamine source is chitin.