Enzymic synthesis of 3'-O- and 6'-O-N-acetylglucosaminyl-N-acetyllactosaminide glycosides catalyzed by beta-N-acetyl-D-hexosaminidase from Nocardia orientalis.

beta-N-acetyl-D-hexosaminidase from Nocardia orientalis catalyzed the synthesis of beta-D-GlcNAc-(1 --> 3)-beta-D-Gal-(1 --> 4)-beta-D-GlcNAc-OC6H4NO2-p (1) and beta-D-GlcNAc-(1 --> 6)-beta-D-Gal-(1 --> 4)-beta-D-GlcNAc-OC6H4NO2-p (2) with its isomer beta-D-Gal-(1 --> 4)-[beta-D-GlcNAc-(1 --> 6)]-beta-D-GlcNAc-OC6H4NO2-p (3) through N-acetylglucosaminyl transfer from N-,N'-diacetylchitobiose to p-nitrophenyl beta-N-acetyllactosaminide. The enzyme formed a mixture of trisaccharides 1, 2, and 3 in a ratio of 11:33:56. In the case, when an inclusion complex of p-nitrophenyl beta-N-acetyllactosaminide with alpha-CD was used, compounds 1, 2, and 3 were formed in a molar ratio of 24:63:13. The regioselectivity of glycosidase-catalyzed formation of the trisaccharide glycosides was substantially changed. It resulted not only in a significant increase of the proportion of the desired compounds 1 and 2 but also in the substantial increase of the overall yield of transfer products.