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    • 6. 发明授权
    • Method for isolation of aminoacylase
    • 分离氨基酰化酶的方法
    • US4532214A
    • 1985-07-30
    • US384766
    • 1982-06-03
    • Bela SzajaniJanosne KissJozsefne IvonyIren HuberLaszlo BorosIvan Daroczi
    • Bela SzajaniJanosne KissJozsefne IvonyIren HuberLaszlo BorosIvan Daroczi
    • C12N9/80C12N11/08C12P13/12C12P41/00
    • C12P41/007C12N11/08C12N9/80C12P13/12Y10S435/814Y10S435/815Y10S435/816
    • Aminoacylase is isolated from mammal kidneys by comminuting and homogenizing mammal kidney in water, centrifuging to form an aqueous extract, heating the aqueous extract at 60.degree. to 80.degree. C. for 5 to 15 minutes, centrifuging, adding a salt such as ammonium sulfate to the resultant supernatant, centrifuging to separate solids, dissolving the solids in water, dialyzing and recovering active aminoacylase from the dialyzed solution. The process produces aminoacylase with high specific activity and the process requires fewer purification steps. The aminoacylase obtained may be utilized directly without any subsequent purification to produce immonobilized aminoacylase. Immobilization can be carried out by covalent bonding of the aminoacylase to a partially hydrolyzed Akrilex P type acryl amide-N,N'-methylene-bis(arylamide) copolymer. The recovered aminoacylase can be subjected to two chromatographic purification steps to produce a very pure aminoacylase.
    • 通过在哺乳动物肾脏中粉碎和匀浆哺乳动物肾脏,离心分离出氨基酰化酶,离心分离形成水提取物,将其在60-80℃加热5〜15分钟,离心分离,加入硫酸铵等盐 生成的上清液,离心分离固体,将固体溶解在水中,透析并从透析溶液中回收活性氨基酰化酶。 该方法产生具有高比活性的氨基酰化酶,该方法需要较少的纯化步骤。 所获得的氨基酰化酶可以直接使用而无需任何后续的纯化以产生单克隆的氨基酰化酶。 可以通过氨基酰化酶与部分水解的Akrilex P型丙烯酰胺-N,N'-亚甲基 - 双(芳基酰胺)共聚物共价键来进行固定。 回收的氨基酰化酶可以进行两个色谱纯化步骤以产生非常纯的氨基酰化酶。