PENICILLIN V AMIDOHYDROLASE GENE FROM FUSARIUM OXYSPORUM FOR ENZYMATIC HYDROLYSIS OF PENICILLIN V (PHENOXY-METHYLPENICILLIN) INTO 6-AMINOPENICILLANIC ACID (6-APA) THAT IS THE ACTIVE BETA-LACTAM NUCLEUS USED IN THE MANUFACTURE OF SEMI-SYNTHETIC PENICILLINS

摘要

PURPOSE: A Penicillin V amidohydrolase gene from Fusarium oxysporum is provided, which enzyme penicillin V amidohydrolase is used for enzymatic hydrolysis of penicillin V (phenoxy-methylpenicillin) into 6-aminopenicillanic acid (6-APA) that is the active beta-lactam nucleus used in the manufacture of semi-synthetic penicillins, and is firstly isolated and purified from Fusarium oxysporum. CONSTITUTION: The expression vector comprises the nucleotide sequence encoding the amino acid sequences of SEQ ID NO:19 or SEQ ID NO:22, wherein the expression vector comprises a replication origin, a promoter and a transcription termination sequence; and the expression vector comprises phleomycin resistant gene as a selection marker. The expression vector comprises the nucleotide sequence encoding the amino acid sequences of SEQ ID NO:18 or SEQ ID NO:21, wherein the expression vector comprises a promoter having the nucleotide sequence of SEQ ID NO:23; the expression vector comprises a replication origin and a structure protein; the expression vector comprises phleomycin resistant gene as a selection marker; and the expression vector is pBMFXPVA6 shown in figure 10, pBMFXPVA7 shown in figure 11, pBMPVA-P shown in figure 14 or pBMPVA-M shown in figure 16. A host cell comprises the expression vector comprising the nucleotide sequence encoding the amino acid sequences of SEQ ID NO:19 or SEQ ID NO:22 or the expression vector comprising the nucleotide sequence encoding the amino acid sequences of SEQ ID NO:18 or SEQ ID NO:21, wherein the host cell is Fusarium oxysporum.