Synthesis of β-d-fructofuranosyl-(2→1)-2-acetamido-2-deoxy-α-d-glucopyranoside (N-acetylsucrosamine) using β-fructofuranosidase-containing Aspergillus oryzae mycelia as a whole-cell catalyst

[Display omitted] ► N-Acetylsucrosamine was synthesized using Aspergillus oryzae mycelia as a whole-cell catalyst. ► The enzyme involved in the synthesis of N-acetylsucrosamine was identified. ► Our method represents a low-cost and easy alternative for the production of N-acetylsucrosamine. ► N-acetylsucrosamine is more stable in acidic solution than is sucrose. Using soft granules consisting of Celite 535 and dried Aspergillus oryzae NBRC100959 mycelia containing β-fructofuranosidase as a whole-cell catalyst, N-acetylsucrosamine [β-d-fructofuranosyl-(2→1)-2-acetamido-2-deoxy-α-d-glucopyranoside] was produced from sucrose and 2-acetamido-2-deoxy-d-glucose by enzymatic transfructosylation. The isolated yield of N-acetylsucrosamine from the reaction mixture was 22.1% (from sucrose). The result of N-terminal amino acid sequence analysis indicated that the enzyme involved in the synthesis of N-acetylsucrosamine is a product from gene (NCBI accession number; NW_001884675, locus tag; AOR_1_1114084) encoding putative β-fructofuranosidase on chromosome 6 of strain NBRC100959. The N-acetylsucrosamine we produced is highly soluble in water and is more stable in acidic solution than sucrose. The disaccharide was also produced using dried mycelia prepared from another A. oryzae strains.