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    • 25. 发明授权
    • Methods of producing doxorubicin
    • 生产多柔比星的方法
    • US5976830A
    • 1999-11-02
    • US653650
    • 1996-05-24
    • William R. StrohlMichael L. DickensCharles L. Desanti
    • William R. StrohlMichael L. DickensCharles L. Desanti
    • C12N9/02C12N15/52C12N15/76C12P19/56C12P1/00C07H21/04
    • C12N9/0077C12N15/52C12N15/76C12P19/56
    • The present invention provides novel methods for producing doxorubicin using daunomycin as a substrate. One method employs a genetically engineered host microorganism which is transformed with a vector, preferably a plasmid, which contains the doxA gene. Preferably, the doxA gene, also referred to herein as "doxA", is cloned into a plasmid which is then introduced into the host microorganism, preferably a bacterial host, more preferably Streptomyces, to provide a transformed host microorganism. The doxA gene, when present on a plasmid, confers on the transformed host the ability to convert daunomycin and 13-dihydrodaunomycin, to doxorubicin. The doxA gene encodes a P450-like enzyme which catalyzes the hydroxylation of daunomycin and 13-dihydrodaunomycin at C-14 to form doxorubicin; such enzyme is designated "daunomycin C-14 hydroxylase". Thus, the expression of doxA in the transformed host using a plasmid which contains doxA enables the transformed host to convert daunomycin to doxorubicin. The doxorubicin is then extracted from host microorganism cultures.
    • 本发明提供使用道诺霉素作为底物生产多柔比星的新方法。 一种方法采用遗传工程的宿主微生物,其用含有doxA基因的载体,优选质粒转化。 优选地,也在本文中称为“doxA”的doxA基因被克隆到质粒中,然后将其导入宿主微生物,优选细菌宿主,更优选链霉菌,以提供转化的宿主微生物。 当存在于质粒上时,doxA基因赋予转化的宿主将道诺霉素和13-二氢青霉素转化为多柔比星的能力。 doxA基因编码P450样酶,其催化道诺霉素和13-二氢青霉素在C-14的羟基化形成多柔比星; 这种酶被称为“道诺霉素C-14羟化酶”。 因此,使用含有doxA的质粒,转化宿主中doxA的表达使转化的宿主能够将道诺霉素转化为多柔比星。 然后从宿主微生物培养物中提取多柔比星。