Patent ID: 8846373
Filing Date: 2014-09-30
Classification: C12N,C12P

Abstract:
1. An isolated eukaryotic cell which is modified to comprise at least one nucleic acid encoding a polypeptide, GlcNAc-6-P 2′-epimerase (GNPE), which is capable of converting N-acetyl-D-glucosamine-6-phosphate (GlcNAc-6-P) to N-acetyl-D-mannosamine-6-phosphate (ManNAc-6-P), wherein each nucleic acid encoding said polypeptide is operably-linked to a promoter functional in said cell; wherein said nucleic acid is obtained from a source belonging to the domain Bacteria; and wherein said polypeptide converts intracellular GlcNAc-6-P to intracellular ManNAc-6-P in an intact cell; and wherein said polypeptide is selected from the group consisting of: (a) a polypeptide comprising SEQ ID NO: 6; (b) a variant of the polypeptide specified in (a) that is at least 90% identical to SEQ ID NO: 6 in which GNPE function is retained; (c) a variant of the polypeptide specified in (b) that contains conservative amino acid substitutions in which GNPE function is retained; (d) a truncated or fusion variant of the polypeptide specified in (a), (b), or (c) comprising one or more insertions or deletions of amino acids in which GNPE function is retained; (e) a truncated or fusion variant of the polypeptide specified in (a), (b), (c), or (d) comprising one or more insertions or deletions of amino acids in which GNPE function is retained; wherein said modified cell also comprises at least one nucleic acid encoding a polypeptide (f) sialic acid-9-phosphate synthase (SAS) [Neu5Ac-9P synthetase, NANS], which is capable of converting ManNAc-6-P to N-acetylneuraminate-9-P (Neu5Ac-9P);