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    • 25. 发明申请
    • ENZYME EXTRACTION AND PURIFICATION PROCESSES
    • 酶提取和纯化方法
    • WO1984004329A1
    • 1984-11-08
    • PCT/AU1984000069
    • 1984-04-19
    • LA TROBE UNIVERSITYSCOPES, Robert, Kerry
    • LA TROBE UNIVERSITY
    • C12N09/12
    • C12N9/0006C12N9/00C12N9/12Y10S435/814
    • A process for the preparation of an enzyme extract containing glucose 6-phosphate dehydrogenase, glucokinase, pyruvate kinase and fructokinase, derived from microorganism cells, by subjecting Zymomonas mobilis bacterium cells to extraction with an extraction medium comprising a partially water-miscible organic solvent; a non-ionic surfactant; and lysozyme; under neutral to alkaline pH conditions to provide an extract containing said enzymes. The process can be integrated with a process for the sequential isolation of the glucose 6-phosphate dehydrogenase, glucokinase, pyruvate kinase and fructokinase, present in the enzyme extract, by sequentially contacting the extract in a buffer with a plurality of affinity chromatography adsorbents each of which is a protein-binding dye bound to a support matrix and selective with respect to said enzymes so that glucose 6-phosphate dehydrogenase and glucokinase are isolated from the extract by the correspondingly selective adsorbent, followed by isolation of pyruvate kinase and fructokinase by the correspondingly selective adsorbents, then elution to recover each enzyme from the correspondingly selective adsorbent of said plurality of adsorbents, with optional purification of each enzyme.
    • 一种制备含有来自微生物细胞的葡萄糖-6-磷酸脱氢酶,葡萄糖激酶,丙酮酸激酶和果糖激酶的酶提取物的方法,通过使运动发酵单胞菌细胞用含有部分水混溶性有机溶剂的提取介质进行提取; 非离子表面活性剂; 和溶菌酶; 在中性至碱性pH条件下提供含有所述酶的提取物。 该方法可以与用于顺序分离存在于酶提取物中的葡萄糖-6-磷酸脱氢酶,葡萄糖激酶,丙酮酸激酶和果糖激酶的方法,通过将缓冲液中的提取物与多个亲和层析吸附剂顺序接触, 其是与支持基质结合的蛋白质结合染料并且相对于所述酶具有选择性,从而通过相应的选择性吸附剂从提取物中分离葡萄糖-6-磷酸脱氢酶和葡萄糖激酶,然后相应地分离丙酮酸激酶和果糖激酶 选择性吸附剂,然后洗脱以从所述多个吸附剂的相应选择性吸附剂中回收每种酶,并可任选地纯化每种酶。