Sugimoto Kazuhisa, Nomura Koji, Nishiura Hiromi, Ohdan Kohji, Ohdan Kohji, Hayashi Hideo, Kuriki Takashi
Biochemical Research Laboratory, Ezaki Glico Co Ltd, 4-6-5 Utajima, Osaka, Japan.
J Biosci Bioeng. 2007 Jul;104(1):22-9. doi: 10.1263/jbb.104.22.
We examined the synthesis of benzoyl glucoside using the transglucosylation reaction of sucrose phosphorylase. Sucrose phosphorylase from Streptococcus mutans showed marked transglucosylating activity, particularly under acidic conditions. On the other hand, sucrose phosphorylase from Leuconostoc mesenteroides showed very weak transglucosylating activity. Three main products were detected from the reaction mixture using benzoic acid as an acceptor molecule and sucrose as a donor molecule. These compounds were identified as 1-O-benzoyl alpha-D-glucopyranoside, 2-O-benzoyl alpha-D-glucopyranose and 2-O-benzoyl beta-D-glucopyranose on the basis of their isolation and the isolation of their acetylated products and subsequent analysis using 1D- and 2D-NMR analyses. From the results of the time-course analyses of the enzyme reaction and the degradation of 1-O-benzoyl alpha-D-glucopyranoside, 1-O-benzoyl alpha-D-glucopyranoside was considered to be initially produced by the transglucosylation reaction of the enzyme, and 2-O-benzoyl alpha-D-glucopyranose and 2-O-benzoyl beta-D-glucopyranose were produced from 1-O-benzoyl alpha-D-glucopyranoside by intramolecular acyl migration reaction. The acceptor specificity in the glucosylation reaction of S. mutans sucrose phosphorylase was also investigated. This sucrose phosphorylase could transglucosylate toward various carboxylic compounds. Short-chain fatty acids, hydroxy acids and dicarboxylic acids were also glucosylated with this sucrose phosphorylase.
我们利用蔗糖磷酸化酶的转糖基化反应研究了苯甲酰葡萄糖苷的合成。变形链球菌的蔗糖磷酸化酶表现出显著的转糖基化活性，尤其是在酸性条件下。另一方面，肠膜明串珠菌的蔗糖磷酸化酶表现出非常弱的转糖基化活性。以苯甲酸为受体分子、蔗糖为供体分子，从反应混合物中检测到三种主要产物。基于它们及其乙酰化产物的分离以及随后使用一维和二维核磁共振分析进行的分析，这些化合物被鉴定为1-O-苯甲酰基-α-D-吡喃葡萄糖苷、2-O-苯甲酰基-α-D-吡喃葡萄糖和2-O-苯甲酰基-β-D-吡喃葡萄糖。根据酶反应的时间进程分析结果以及1-O-苯甲酰基-α-D-吡喃葡萄糖苷的降解情况，认为1-O-苯甲酰基-α-D-吡喃葡萄糖苷最初是由该酶的转糖基化反应产生的，而2-O-苯甲酰基-α-D-吡喃葡萄糖和2-O-苯甲酰基-β-D-吡喃葡萄糖是由1-O-苯甲酰基-α-D-吡喃葡萄糖苷通过分子内酰基迁移反应产生的。还研究了变形链球菌蔗糖磷酸化酶糖基化反应中的受体特异性。这种蔗糖磷酸化酶可以将糖基转移到各种羧酸化合物上。短链脂肪酸、羟基酸和二羧酸也能用这种蔗糖磷酸化酶进行糖基化。
