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    • 20. 发明授权
    • 섬유소결합도메인을 이용한 활성형 단백질 응집체 입자의 제조방법
    • 使用纤维素结合域生产活性蛋白复合颗粒的制备方法
    • KR100776344B1
    • 2007-11-15
    • KR1020060057224
    • 2006-06-26
    • 한국생명공학연구원
    • 이승구송재준반재구정흥채최수림
    • C12N15/64
    • A method for production of active protein complex particles by using a cellulose binding domain is provided to produce the active protein complex particles easily and rapidly by expressing in Escherichia coli without pretreatment or immobilization, recover and recycle the particles by centrifugation, and increase immobilization rate by treating with a crosslinking agent. A method for production of active protein complex particles comprises the steps of: preparing a recombinant vector containing a gene encoding cellulose binding domain derived from Cellulomonas fimi having the amino acid sequence of SEQ ID NO:1 and an active protein gene; transforming a host cell with the recombinant vector; and culturing the transformed microorganism, wherein the cellulose binding domain is exoglucanase; the active protein is beta-glucuronidase(gusA) or beta-glucosidase(bglA); the host cell is Escherichia coli; and the active protein complex particles are immobilized by adding a crosslinking agent such as genipin or glutaraldehyde into the particles.
    • 提供通过使用纤维素结合结构域生产活性蛋白质复合物颗粒的方法,通过在大肠杆菌中表达而不经预处理或固定,通过离心回收和再循环颗粒,容易且快速地制备活性蛋白质复合物颗粒,并通过 用交联剂处理。 制备活性蛋白质复合物颗粒的方法包括以下步骤:制备含有编码具有SEQ ID NO:1的氨基酸序列的纤维单胞菌的纤维素结合结构域的基因和活性蛋白质基因的重组载体; 用重组载体转化宿主细胞; 培养转化的微生物,其中纤维素结合域为外切葡聚糖酶; 活性蛋白是β-葡糖苷酸酶(gusA)或β-葡糖苷酶(bglA); 宿主细胞是大肠杆菌; 并通过向颗粒中加入京尼平或戊二醛等交联剂来固定活性蛋白质复合物颗粒。