发明公开
EP0969088A1 ENZYMATIC PROCESS FOR THE PREPARATION OF CEPHALOSPORANIC 7$b(g)-(4-CARBOXYBUTANAMIDE) ACID BY MEANS OF THE MODIFIED ENZYME D-AMINOACID OXIDASE OF $i(TRIGONOPSIS VARIABILIS) PRODUCED IN $i(ESCHERICHIA COLI)
审中-公开
酶促过程用于产生修正后的dAMINOSÄUREOXIDASEENZYMS头孢-7-β-4-羧丁基酰胺MEANS在大肠杆菌中生产什么
- 专利标题： ENZYMATIC PROCESS FOR THE PREPARATION OF CEPHALOSPORANIC 7$b(g)-(4-CARBOXYBUTANAMIDE) ACID BY MEANS OF THE MODIFIED ENZYME D-AMINOACID OXIDASE OF $i(TRIGONOPSIS VARIABILIS) PRODUCED IN $i(ESCHERICHIA COLI)
- 专利标题（中）： 酶促过程用于产生修正后的dAMINOSÄUREOXIDASEENZYMS头孢-7-β-4-羧丁基酰胺MEANS在大肠杆菌中生产什么
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申请号： EP98943898.1申请日： 1998-09-24
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公开(公告)号： EP0969088A1公开(公告)日： 2000-01-05
- 发明人: GARCIA LOPEZ, José Luis , CORTES RUBIO, Estrella , ALONSO PALACIOS, Jorge , MELLADO DURAN, Encarnación , DIEZ GARCIA, Bruno , GUISAN SEIJAS, José Manuel , SALTO MALDONADO, Francisco , BARREDO FUENTE, José Luis
- 申请人： Antibioticos, S.A.
- 申请人地址： Avenida Antibioticos, 59/61, 24089 Léon ES
- 专利权人： Antibioticos, S.A.
- 当前专利权人： Antibioticos, S.A.
- 当前专利权人地址： Avenida Antibioticos, 59/61, 24089 Léon ES
- 代理机构： Elzaburu, Alberto
- 优先权： ES9702008 19970925
- 国际公布： WO9915632 19990401
- 主分类号： C12N9/06
- IPC分类号： C12N9/06
摘要：
Enzymatic process for the preparation of cephalosporanic 7β-(4-carboxybutanamide) acid by using the modified enzyme D-aminoacid oxidase of Trigonopsis variabilis produced in Escherichia coli . The process for the expression of the enzyme comprises: (I) isolating the DNA corresponding to the gene which codes for the enzyme D-aminoacide oxidase; (II) removing the intron which is contained in said gene; (III) inserting the DNA fragment obtained into the plasmide which is capable of replication in Escherichia coli ; (IV) fusing at the extremity 5' of the structural region of the gene a synthetic assembler which contains a nucleotide sequence which codes for six histidines; (V) transforming a strain of Escherichia coli with the resulting recombinant plasmide; (VI) cultivating the transformed cells of Escherichia coli ; and (VII) recovering the enzyme D-aminoacid oxidase of the former cultivation operation through affinity chromatography.
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