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

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

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...

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Hauptverfasser: SALTO MALDONADO FRANCISO, MELLADO DURAN ENCARNACION, DIEZ GARCIA BRUNO, GUISAN SEIJAS JOSE MANUEL, ALONSO PALACIOS JORGE, FUENTE BARREDO, GARCIA LOPEZ JOSE LUIS, CORTES RUBIO ESTRELLA
Format: Patent
Sprache:eng
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BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE
METALLURGY
MICROBIOLOGY
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MUTATION OR GENETIC ENGINEERING
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Zusammenfassung: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.