Details

Autor(en) / Beteiligte

• Matsuo, I. (Meiji Inst. of Health Science, Odawara, Kanagawa (Japan))
• Kim, S
• Yamamoto, Y
• Ajisaka, K
• Maruyama, J
• Nakajima, H
• Kitamoto, K

Titel

Cloning and overexpression of beta-N-acetylglucosaminidase encoding gene nagA from Aspergillus oryzae and enzyme-catalyzed synthesis of human milk oligosaccharide

Ist Teil von

• Bioscience, biotechnology, and biochemistry, 2003-03, Vol.67 (3), p.646-650

Ort / Verlag

Tokyo: Japan Society for Bioscience, Biotechnology, and Agrochemistry

Erscheinungsjahr

2003

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• We isolated a β-N-acetylglucosaminidase encoding gene from the filamentous fungus Aspergillus oryzae, and designated it nagA. The nagA gene encoded a polypeptide of 600 amino acids with significant similarity to glucosaminidases and hexosaminidases of various eukaryotes. A. oryzae strain carrying the nagA gene under the control of the improved glaA promoter produced large amounts of β-N-acetylglucosaminidase in a wheat bran solid culture. The β-N-acetylglucosaminidase was purified from crude extracts of the solid culture by column chromatographies on Q-Sepharose and Sephacryl S-200. This enzyme was used for synthesis of lacto-N-triose II, which is contained in human milk. By reverse hydrolysis reaction, lacto-N-triose II and its positional isomer were synthesized from lactose and D-N-acetylglucosamine in 0.21% and 0.15% yield, respectively.