发明名称 |
Enzymatic process for the preparation of cephalosporanic 7 beta -(4-Carboxybutanamide) acid by means of the modified enzyme D-aminoacid oxidase of trigonopsis variabilis produced in escherichia coli |
摘要 |
Enzymatic process for the preparation of cephalosporanic 7beta-(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 gene which codes for the enzyme D-aminoacid 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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申请公布号 |
US6635458(B2) |
申请公布日期 |
2003.10.21 |
申请号 |
US19990308683 |
申请日期 |
1999.07.15 |
申请人 |
ANTIBIOTICOS, S.A.U. |
发明人 |
GARCIA LOPEZ JOSE LUIS;CORTES RUBIO ESTRELLA;ALONSO PALACIOS JORGE;MELLADO DURAN ENCARNACION;DIEZ GARCIA BRUNO;GUISAN SEIJAS JOSE MANUEL;SALTO MALDONADO FRANCISO;FUENTE BARREDO |
分类号 |
C12N15/09;C12N1/21;C12N9/06;C12P17/18;C12P35/06;C12R1/01;(IPC1-7):C12N9/04;C12N1/20;C12N15/00;C12N5/00;C07K1/00 |
主分类号 |
C12N15/09 |
代理机构 |
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地址 |
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