STREPTOMYCES PEUCETIUS DAUNORUBICIN BIOSYNTHESIS GENE, DNRF - SEQUENCE AND HETEROLOGOUS EXPRESSION
STREPTOMYCES PEUCETIUS DAUNORUBICIN BIOSYNTHESIS GENE, DNRF - SEQUENCE AND HETEROLOGOUS EXPRESSION
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DOI：
10.1099/13500872-141-4-1007
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发表时间：
1995-04-01
期刊：
影响因子：
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通讯作者：
GAROFANO, L
中科院分区：
文献类型：
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作者：
FILIPPINI, S;SOLINAS, MM;GAROFANO, L
在peucetius链霉菌（Streptomyces peucetius） ATCC 29050的柔红霉素（daunorubicin, Dnr）基因簇中发现了dnrF基因，该基因通过C-11羟基化将aklavinone转化为epsilon-rhodomycinone，与其中一个蒽环类耐药基因drrAB接近。dnrF基因被测序，编码一个489个氨基酸的蛋白，分子量为52 kDa，推断的dnrF蛋白与细菌FAD和nadph依赖的羟化酶有显著的相似性，这些羟化酶需要将羟基引入多环芳香族聚酮抗生素或参与芳香族化合物的分解代谢。dnrF在lividans链霉菌TK23和大肠杆菌中的异源表达表明，该基因编码nadph依赖的羟化酶，催化阿克拉维酮羟基化生成epsilon-rhodomycinone。该酶对C-7位糖基化的蒽环类药物无活性，并且随着其他底物修饰，其活性有不同程度的降低，表明DnrF具有显著的底物特异性。
The dnrF gene, responsible for conversion of aklavinone to epsilon-rhodomycinone via C-11 hydroxylation, was mapped in the daunorubicin (Dnr) gene cluster of Streptomyces peucetius ATCC 29050, close to drrAB, one of the anthracycline-resistance genes. The dnrF gene was sequenced and should encode a protein of 489 amino acids with a molecular mass of 52 kDa, The deduced DnrF protein shows significant similarities with bacterial FAD- and NADPH-dependent hydroxylases either required to introduce hydroxyl groups into polycyclic aromatic polyketide antibiotics or involved in catabolism of aromatic compounds. Heterologous expression of dnrF in Streptomyces lividans TK23 and in Escherichia coli demonstrated that the gene encodes a NADPH-dependent hydroxylase catalysing the hydroxylation of aklavinone to yield epsilon-rhodomycinone. The enzyme is inactive on anthracyclines glycosylated at position C-7 and its activity decreases to a different extent with other substrate modifications, indicating that DnrF has a significant substrate specificity.