通过刀豆中N-乙酰己糖胺酶的逆反应酶促合成N-乙酰葡糖胺基环糊精。

Enzymatic synthesis of N-acetylglucosaminyl-cyclodextrin by the reverse reaction of N-acetylhexosaminidase from jack bean.

作者信息

Hamayasu K, Fujita K, Hara K, Hashimoto H, Tanimoto T, Koizumi K, Nakano H, Kitahata S

机构信息

Bio Research Corporation of Yokohama, Japan.

出版信息

Biosci Biotechnol Biochem. 1999 Oct;63(10):1677-83. doi: 10.1271/bbb.63.1677.

DOI:10.1271/bbb.63.1677

PMID:10586495

Abstract

Novel heterobranched cyclodextrins (CDs), N-acetylglucosaminyl-cyclodextrins (GlcNAc-CD), were synthesized from a mixture of GlcNAc and alpha, beta, or gamma CD by the reverse reaction of N-acetylhexosaminidase from jack bean. Optimum pH and temperature for the production of GlcNAc-alpha CD by N-acetylhexosaminidase were pH 4.9 and 50-70 degrees C, respectively. The maximum yield of GlcNAc-alpha CD was 17.5% (mol/mol) at the concentration of 1 M GlcNAc and 0.25 M alpha CD. The reverse reaction product, GlcNAc-alpha CD, was separated into two peaks by HPLC analysis on the ODS column. Their structures were identified as 6-O-beta-D-N-acetylglucosaminyl-alpha CD and 2-O-beta-D-N-acetylglucosaminyl-alpha CD by FAB-MS and NMR spectroscopies. N-Acetylhexosaminidase from jack bean also synthesized N-acetylgalactosaminyl-alpha CD from N-acetylgalactosamine and alpha CD.

摘要

新型杂支链环糊精（CDs），即N-乙酰葡糖胺基环糊精（GlcNAc-CD），是通过刀豆中的N-乙酰己糖胺酶的逆反应，由GlcNAc与α、β或γ环糊精的混合物合成的。N-乙酰己糖胺酶生产GlcNAc-α CD的最适pH和温度分别为pH 4.9和50 - 70℃。在1 M GlcNAc和0.25 M α环糊精的浓度下，GlcNAc-α CD的最大产率为17.5%（摩尔/摩尔）。通过ODS柱上的HPLC分析，逆反应产物GlcNAc-α CD被分离为两个峰。通过快原子轰击质谱（FAB-MS）和核磁共振光谱（NMR）确定它们的结构分别为6-O-β-D-N-乙酰葡糖胺基-α CD和2-O-β-D-N-乙酰葡糖胺基-α CD。刀豆中的N-乙酰己糖胺酶还从N-乙酰半乳糖胺和α环糊精合成了N-乙酰半乳糖胺基-α CD。