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    • 7. 发明公开
    • 퓨조박테리움 뉴클레텀 유래의 신규 글리코실트랜스퍼라제및 이를 이용한 올리고당 제조방법
    • 来自金丝桃核的新型糖苷酶及其用途
    • KR1020080049490A
    • 2008-06-04
    • KR1020060120115
    • 2006-11-30
    • 한국생명공학연구원
    • 강현아김성훈오두병권오석정재갑
    • C12N9/24
    • A glycosyltransferase derived from Fusobacterium nucleatum is provided to be involved with synthesis of lipo-oligosaccharide, lipo-polysaccharide, and mucous polysaccharide oligosaccharide and show excellent expression efficiency, very high active type protein production yield and excellent enzymatic activity when produced into a recombinant protein form in a microorganism such as E. coli and yeast. A protein having the activity of glycosyltransferase derived from Fusobacterium nucleatum is characterized in that a saccharide is added to a receptor using UDP-Glc(uridine-5'-diphospho-glucose) or UDP-Gal(uridine-5'-diphospho-galactose) as a donor. An expression vector includes a gene encoding the protein. A transformant transformed by the expression vector is Origami(DE3)pLysS/pET-Fnu_GT(deposition no. KCTC11026BP). A method for preparing a protein having a recombinant glycosyl transferase activity comprises the steps of: (a) preparing a recombinant expression vector including a gene encoding glycosyltransferase derived from Fusobacterium; (b) transforming a host cell using the recombinant expression vector; and (c) culturing the transformant. A method for preparing a sugar chain precursor, a glycoprotein, and glycolipid specific to glycosylation comprises a step of contacting the protein having the glycosylase activity with a mixture of a receptor and a donor of the glycosylation.
    • 提供源自核梭菌的糖基转移酶涉及脂多糖,脂多糖和粘多糖寡糖的合成,并且当产生重组蛋白质形式时,表现出优异的表达效率,非常高的活性型蛋白质产生产率和优异的酶活性 在微生物如大肠杆菌和酵母中。 具有衍生自核棒状杆菌的糖基转移酶活性的蛋白质的特征在于,使用UDP-Glc(尿苷-5'-二磷酸 - 葡萄糖)或UDP-Gal(尿苷-5'-二磷酸 - 半乳糖)向受体中加入糖类, 作为捐助者。 表达载体包括编码蛋白质的基因。 由表达载体转化的转化体是折纸(DE3)pLysS / pET-Fnu_GT(沉积编号KCTC11026BP)。 一种制备具有重组糖基转移酶活性的蛋白质的方法包括以下步骤:(a)制备包含编码来自Fusobacterium的糖基转移酶的基因的重组表达载体; (b)使用重组表达载体转化宿主细胞; 和(c)培养转化体。 用于制备糖基化特异性的糖链前体,糖蛋白和糖脂的方法包括使具有糖基化酶活性的蛋白质与糖基化受体和供体的混合物接触的步骤。