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    • 10. 发明授权
    • Cephalosporin C acylase
    • 头孢菌素C酰基转移酶
    • US5336613A
    • 1994-08-09
    • US19870
    • 1993-02-19
    • Mineo NiwaYoshimasa SaitoHitoshi SasakiYoshinori Ishii
    • Mineo NiwaYoshimasa SaitoHitoshi SasakiYoshinori Ishii
    • C12N1/21C12N9/80C12N15/09C12N15/55C12P35/02C12P35/06C12R1/19C12N15/52
    • C12P35/02C12N9/80
    • The present invention concerns a mutant cephalosporin C acylase derived from a precursor of the formula:A.sup.1-268 --X.sup.1 --Tyr--X.sup.2 --A.sup.272-304 --X.sup.3 --A.sup.306-773(SEQ ID NO:1), wherein:A.sup.1-268 is the same amino acid sequence as that from Thr.sup.1 to Gly.sup.268 of native CC acylase,A.sup.272-304 is the same amino acid sequence as that from Gln.sup.272 to Tyr.sup.304 of native CC acylase,A.sup.306-773 is the same amino acid sequence as that from Val.sup.306 to Ala.sup.773 of native CC acylase,X.sup.1 is Met or other amino acid,X.sup.2 is Ala or Tyr, andX.sup.3 is Cys or Ser,provided that when X.sup.1 is Met and X.sup.2 is Ala, X.sup.3 is Ser; and that the mutant cephalosporin C acylase has a property selected from the group consisting of higher enzymatic potency and higher processing efficiency, as compared to native CC acylase. The present invention also concerns DNA encoding the mutant cephalosporin C acylase, an expression vector containing the DNA, a host cell transformed with the expression vector, a process for producing the mutant cephalosporin C acylase by culturing the transformed host cell, and a process for preparing a cephalosporin C using the mutant cephalosporin C acylase.
    • 本发明涉及衍生自下式的前体的突变型头孢菌素C酰基转移酶:A1-268-X1-Tyr-X2-A272-304-X3-A306-773(SEQ ID NO:1),其中:A1-268是 与天然CC酰基转移酶A272-304的Thr1至Gly268相同的氨基酸序列与天然CC酰基转移酶从Gln272至Tyr304的氨基酸序列相同,氨基酸序列与Val306至 天然CC酰基转移酶的Ala773,X1是Met或其他氨基酸,X2是Ala或Tyr,X3是Cys或Ser,条件是当X1是Met,X2是Ala时,X3是Ser; 并且与天然CC酰基转移酶相比,突变型头孢菌素C酰基转移酶具有选自较高酶效力和较高加工效率的性质。 本发明还涉及编码突变型头孢菌素C酰基转移酶的DNA,含有DNA的表达载体,用表达载体转化的宿主细胞,通过培养转化的宿主细胞产生突变型头孢菌素C酰基转移酶的方法,以及制备 使用突变型头孢菌素C酰基转移酶的头孢菌素C。