KMID : 1094720000050030215
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Biotechnology and Bioprocess Engineering 2000 Volume.5 No. 3 p.215 ~ p.218
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Production of sialyltrisaccharides using ¥â-galactosidase and trans -sialidase in one pot
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Lee Sun-Gu
Kim Byung-Gee
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Abstract
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Sialyltrisaccharides based on ¥â-galactosyldisaccharides were synthesized using ¥â-galactosidase andtrans-sialidase in one pot. Using ¥â-galactosidase fromBacillus circulans andtrans-sialidase fromTrypanosoma cruzi simultaneously, 6 mM sialyltrisaccharides composed of about 95% NeuAc¥á(2,3)Gal¥â(1,4)GlcNAc and 5% NeuAc¥á(2,3)Gal¥â(1,6)GlcNAc were produced from a reaction mixture containing 25 mM 0-nitrophenyl-¥â-D-galactopyranoside, 100 mM N-acety lglucosamine and 10 mM p-nitrophenyl-¥á-D-N-acetylneuraminic acid. One beauty of this reaction was that a secondary hydrolysis of the disaccharide intermediate occurring between the activated galactopyranoside and N-acetylglucosamine was prevented. Using ¥â-galactosidase fromEscherichia coli and the sametrans-sialidase, 15 mM sialyltrisaccharides composed of about 90% NeuAc¥á(2,3)Gal¥â(1,6)GlcNAc and 10% NeuAc¥á(2,3)Gal¥â (1,4)GlcNAc were produced from a reaction mixture containing 400 mM galactose, 800 mM N-acetylglucosamine and 20 mMp-nitrophenyl-¥á-D-N-acetylneuraminic acid. In this study, the reverse-galactosylation reaction between galactose and N-acetylglucosamine was dominant since the disaccharide intermediate mainly resulted in the sialylated product.
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KEYWORD
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trans-sialidase, ¥â-galactosidase, trans-galactosylation, reverse-galactosylation
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