公开(公告)号：EP0839914A1

公开(公告)日：1998-05-06

申请号：EP97919404.0

申请日：1997-04-18

申请人： Antibioticos, S.A.

发明人： GARCIA LOPEZ, José Luis ,  CORTES RUBIO, Estrella ,  GUISAN SEIJAS, José Manuel ,  BARREDO FUENTE, José Luis ,  DIEZ GARCIA, Bruno ,  COLLADOS DE LA VIEJA, Alfonso ,  VITALLER ALBA, Alejandro ,  SALTO MALDONADO, Francisco

IPC分类号： C12N15/55 ,  C12N9/14 ,  C12N1/21 ,  C12P35/06 ,  C07K1/22

CPC分类号： C12N9/86 ,  C07K2319/00 ,  C12N9/84

摘要： A procedure for producing a modified 7β-(4-carboxybutanamide) cephalosporinase enzyme which can be purified in a single chromatographic step. The procedure for production of the enzyme involves: mutagenizing the gene which codes for the enzyme from Acinetobacter sp . ATCC 53891 by inserting a nucleotide sequence coding for six histidine residues; fusing the mutant gene with high-efficiency promoter DNA sequences; transforming Escherichia coli cells with the fusion gene construct; growing the transformed Escherichia coli cells; and recovering the enzyme by the use of supports which contain metal chelates. 7-Aminocephalosporanic acid is an important intermediate for the manufacture of a wide range of antibacterial agents of the cephalosporin family.

摘要翻译： 一种生产改性7测试程序 - （4-羧丁基酰胺）头孢菌素酶，其可以在单个层析步骤中纯化。 产生酶的过程包括：诱变基因，其编码来自不动杆菌的酶。 ATCC 53891通过插入编码六个组氨酸残基的核苷酸序列; 熔融配合高效率启动子的DNA序列的突变基因; 转化大肠杆菌细胞与所述融合基因构建体; 培育转化的大肠杆菌细胞; 和回收通过使用含有金属螯合物的载体的酶。 7-氨基头孢烷酸是适用范围广的头孢菌素家族的抗菌剂的制造中的重要中间体。

公开(公告)号：EP0843015A1

公开(公告)日：1998-05-20

申请号：EP97919405.7

申请日：1997-04-18

申请人： Antibioticos, S.A.

发明人： GARCIA LOPEZ, José Luis ,  CORTES RUBIO, Estrella ,  ALONSO PALACIOS, Jorge ,  BARREDO FUENTE, José Luis ,  DIEZ GARCIA, Bruno ,  MORENO VALLE, Miguel Angel ,  SCHLEISSNER SANCHEZ, Carmen ,  COLLADOS DE LA VIEJA, Alfonso ,  VITALLER ALBA, Alejandro ,  SALTO MALDONADO, Francisco

IPC分类号： C12N15/53 ,  C12N9/06 ,  C12P35/06 ,  C12N1/21

CPC分类号： C12N9/0024

摘要： Process for producing the enzyme D-amino acid oxydase of Rhodotorula gracilis in host cells. The process for the expression of the enzyme comprises isolating the complementary DNA corresponding to the messager RNA of the gene which codes for the D-amino acid oxydase of any strain of Rhodotorula gracillis producing said enzyme; fusing the fragment of DNA which codes for D-amino acid oxydase of Rhodotorula gracillis with a DNA sequence which contains a site of union to the ribosome and a high efficiency promoter sequence for the express of genes in host cells; inserting the DNA fragment into a plasmid appropriate for the host cell; cultivating the host cells transformed with said plasmid; and collecting the enzyme.

摘要翻译： 在宿主细胞中生产细小红球菌的D-氨基酸氧化酶酶的方法。 表达酶的过程包括分离对应于编码产生所述酶的任何红曲霉菌株的D-氨基酸氧化酶的基因的信使RNA的互补DNA; 将编码红景天细菌的D-氨基酸氧化酶的DNA片段与含有与核糖体结合的位点的DNA序列和用于在宿主细胞中表达基因的高效启动子序列融合; 将DNA片段插入适合宿主细胞的质粒中; 培养用所述质粒转化的宿主细胞; 并收集酶。