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    • 1. 发明授权
    • Enzyme catalysis in carbon dioxide fluids
    • 二氧化碳液体中的酶催化
    • US06211422B1
    • 2001-04-03
    • US09352249
    • 1999-07-13
    • Joseph M. DeSimoneRuben G. CarbonellStephanie A. CretteJonathan L. Kendall
    • Joseph M. DeSimoneRuben G. CarbonellStephanie A. CretteJonathan L. Kendall
    • C12P100
    • C12P41/001C12P1/00Y02P20/544
    • A method for the enzymatic production of a desired product in a liquid or supercritical carbon dioxide fluid. The method comprises: (a) providing a reaction mixture comprising at least one reaction substrate and a crosslinked enzyme crystal catalyst in a liquid or supercritical carbon dioxide fluid; and (b) reacting said substrate with said crosslinked enzyme crystal catalyst in said carbon dioxide fluid to produce said product. The reacting step is typically followed by the step of (c) separating the product from said reaction mixture. The crosslinked enzyme crystal catalyst preferably comprises an enzyme crystal crosslinked with a multifunctional crosslinking agent, said crosslinked enzyme crystal having resistance to exogeneous proteolysis, such that the crosslinked enzyme crystal retains at least about 80 percent of its initial activity after incubation for three hours in the presence of a concentration of Pronase™ protease that causes the soluble uncrosslinked form of the enzyme that is crystallized to form said enzyme crystal that is crosslinked to lose at least about 80 percent of its initial activity under the same conditions. Reaction mixtures useful for carrying out the foregoing method and product mixtures produced by the foregoing method are also disclosed.
    • 在液体或超临界二氧化碳流体中酶促生产所需产物的方法。 该方法包括:(a)在液体或超临界二氧化碳流体中提供包含至少一个反应底物和交联的酶晶体催化剂的反应混合物; 和(b)使所述底物与所述交联的酶晶体催化剂在所述二氧化碳流体中反应以产生所述产物。 反应步骤通常是(c)从所述反应混合物中分离产物的步骤。 交联酶晶体催化剂优选包含与多官能交联剂交联的酶晶体,所述交联酶晶体具有对外源性蛋白水解的抗性,使得交联的酶晶体在孵育3小时后保留其初始活性的至少约80% 存在一种浓度的Pronase TM蛋白酶,其引起可溶性的未交联形式的酶,其被结晶以形成所述交联的酶晶体,以在相同条件下损失其初始活性的至少约80%。 还公开了可用于实施上述方法的反应混合物和通过前述方法制备的产物混合物。