申请人:Seikagaku Corporation
公开号:US08129148B2
公开(公告)日:2012-03-06
Provided are a method for producing a fraction containing more than 50% of CH represented by the general formula (1), which comprises at least the step of allowing a glucuronic acid donor, an N-acetylgalactosamine donor, a saccharide receptor, a chondroitin polymerase derived from the Escherichia coli K4 strain, and Mn2+ at a final concentration of 0.02 to 100 mM to coexist, and performing a reaction thereof under conditions of 20 to 40° C. and pH 6 to 8 for 0.5 minutes to 4 hours, and a method for producing a fraction containing more than 50% of CH represented by the general formula (2), which comprises at least the step of performing the reaction under same conditions for 10 hours or longer, which enable industrial scale production of a CH fraction of a controlled even number saccharide and odd number saccharide content ratio by a simple procedure at a low cost.
(GlcA-GalNAc)n (1)
GalNAc-(GlcA-GalNAc)n (2)
(In the formula, GlcA represents a glucuronic acid residue, GalNCAc represents a N-acetylgalactosamine residue, - represents a glycosidic bond, and n represents an arbitrary integer.)
提供了一种生产含有50%以上由通式(1)表示的CH的方法,该方法至少包括使葡萄糖醛酸供体,N-乙酰半乳糖胺供体,糖受体,源自大肠杆菌K4菌株的软骨素聚合酶和最终浓度为0.02至100 mM的Mn2+共存,并在20至40°C和pH 6至8的条件下进行反应0.5分钟至4小时,以及生产含有50%以上由通式(2)表示的CH的方法,该方法至少包括在相同条件下进行反应10小时或更长时间,从而以低成本的简单程序实现控制偶数糖和奇数糖含量比的CH分数的工业规模生产。(GlcA-GalNAc)n(1)GalNAc-(GlcA-GalNAc)n(2)(在公式中,GlcA表示葡萄糖醛酸残基,GalNCAc表示N-乙酰半乳糖胺残基,-表示糖苷键,n表示任意整数。)