Mutation and cloning of eryG, the structural gene for erythromycin O-methyltransferase from Saccharopolyspora erythraea, and expression of eryG in Escherichia coli
| Autor: | V E Luebke, J S Tuan, T. J. Paulus, L Katz, G. T. Maine, J. P. Dewitt |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
DNA
Bacterial Restriction Mapping Mutant Gene Expression Erythromycin Molecular cloning medicine.disease_cause Microbiology Streptomyces Plasmid Escherichia coli medicine Cloning Molecular Molecular Biology Chromatography High Pressure Liquid biology Structural gene Methyltransferases biochemical phenomena metabolism and nutrition biology.organism_classification Molecular biology Recombinant Proteins Genes Bacterial Mutation Saccharopolyspora erythraea Plasmids Research Article medicine.drug |
| Zdroj: | Journal of Bacteriology. 172:2541-2546 |
| ISSN: | 1098-5530 0021-9193 |
| DOI: | 10.1128/jb.172.5.2541-2546.1990 |
| Popis: | A mutant strain derived by chemical mutagenesis of Saccharopolyspora erythraea (formerly known as Streptomyces erythreus) was isolated that accumulated erythromycin C and, to a lesser extent, its precursor, erythromycin D, with little or no production of erythromycin A or erythromycin B (the 3"-O-methylation products of erythromycin C and D, respectively). This mutant lacked detectable erythromycin O-methyltransferase activity with erythromycin C, erythromycin D, or the analogs 2-norerythromycin C and 2-norerythromycin D as substrates. A 4.5-kilobase DNA fragment from S. erythraea originating approximately 5 kilobases from the erythromycin resistance gene ermE was identified that regenerated the parental phenotype and restored erythromycin O-methyltransferase activity when transformed into the erythromycin O-methyltransferase-negative mutant. Erythromycin O-methyltransferase activity was detected when the 4.5-kilobase fragment was fused to the lacZ promoter and introduced into Escherichia coli. The activity was dependent on the orientation of the DNA relative to lacZ. We have designated this genotype eryG in agreement with Weber et al. (J.M. Weber, B. Schoner, and R. Losick, Gene 75:235-241, 1989). It thus appears that a single enzyme catalyzes all of the 3"-O-methylation reactions of the erythromycin biosynthetic pathway in S. erythraea and that eryG codes for the structural gene of this enzyme. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|