申请人:Sanyo Chemical Industries, Ltd.
公开号:EP2719767A1
公开(公告)日:2014-04-16
The present invention aims to provide a method for producing a polysaccharide with high efficiency using a polysaccharide synthase. The present invention provides a method for producing a polysaccharide, including allowing polysaccharide synthase (B) to act on ribonucleoside diphosphate-monosaccharide (A) shown below to produce a polysaccharide, wherein in 10 to 100% of the duration in which (B) acts on (A), the concentration of ribonucleoside diphosphate in a reaction solution is lower than 100 times an inhibitory concentration IC50 described below against polysaccharide synthase (B).
Inhibitory concentration IC50: a concentration of ribonucleoside diphosphate at which an enzyme activity of polysaccharide synthase (B) is reduced by half while under a condition where (B) has a concentration at which (B) acts on ribonucleoside diphosphate-monosaccharide (A), wherein ribonucleoside diphosphate-monosaccharide (A) is used as a substrate and ribonucleoside diphosphate is used as an inhibitor.
Ribonucleoside diphosphate-monosaccharide (A): a sugar nucleotide in which a proton of at least one hydroxyl group of at least one monosaccharide (a) selected from the group consisting of triose (a-1), tetrose (a-2), pentose (a-3), hexose (a-4), heptose (a-5), and monosaccharide (a-6) described below is substituted with a functional group represented by any one of chemical formulae (1) to (5) below.
Monosaccharide (a-6) : a monosaccharide selected from the group consisting of (a-1), (a-2), (a-3), (a-4), and (a-5), in which at least one member selected from the group consisting of a proton, a hydroxyl group, and a hydroxymethyl group of the monosaccharide is substituted with substituent (E) described below.
Substituent (E) is at least one substituent selected from the group consisting of carboxyl, amino, N-acetylamino, sulfate, methylester, N-glycolyl, methyl, 1,2,3-trihydroxypropyl, phosphate, and 2-carboxy-2-hydroxyethyl groups.
本发明旨在提供一种利用多糖合成酶高效生产多糖的方法。本发明提供了一种生产多糖的方法,包括让多糖合成酶(B)作用于下图所示的核糖核苷二磷酸-单糖(A)以生产多糖,其中在(B)作用于(A)的10至100%的持续时间内,反应溶液中核糖核苷二磷酸的浓度低于下述对多糖合成酶(B)的抑制浓度IC50的100倍。
抑制浓度 IC50:在(B)对核糖核苷二磷酸单糖(A)起作用的浓度条件下,多糖合成酶(B)的酶活性降低一半的核糖核苷二磷酸浓度,其中核糖核苷二磷酸单糖(A)用作底物,核糖核苷二磷酸用作抑制剂。
核糖核苷二磷酸-单糖 (A):一种糖核苷酸,其中至少一种单糖 (a) 的至少一个羟基的质子被下述化学式 (1) 至 (5) 中任一个所代表的官能团取代,该单糖选自三糖 (a-1)、四糖 (a-2)、戊糖 (a-3)、己糖 (a-4)、庚糖 (a-5) 和下述单糖 (a-6)。
单糖(a-6):选自(a-1)、(a-2)、(a-3)、(a-4)和(a-5)组成的组的单糖,其中至少一个选自质子、羟基和羟甲基组成的组的成员被下述取代基(E)取代。
取代基(E)是选自羧基、氨基、N-乙酰氨基、硫酸基、甲基酯、N-甘氨酰、甲基、1,2,3-三羟基丙基、磷酸基和 2-羧基-2-羟乙基组成的组中的至少一个取代基。