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首页> 外文期刊>Biotechnology Letters >An approach for cloning biosynthetic genes of 2-deoxystreptamine-containing aminocyclitol antibiotics: Isolation of a biosynthetic gene cluster of tobramycin from Streptomyces tenebrarius
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An approach for cloning biosynthetic genes of 2-deoxystreptamine-containing aminocyclitol antibiotics: Isolation of a biosynthetic gene cluster of tobramycin from Streptomyces tenebrarius

机译：一种克隆含2-脱氧链胺胺的氨基环醇抗生素的生物合成基因的方法：从黄链霉菌中分离妥布霉素的生物合成基因簇

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摘要

Genes homologous to 2-deoxystreptamine (DOS) biosynthetic genes were isolated from aminoglycoside producers, Micromonospora and Streptomyces spp., using PCR primers based on the core sequences of 2-deoxyscylloinosose (DOI) synthase and L-glutamine: scyllo-inosose aminotransferase (GIA). Identities of 40 - 45% were observed for DOI synthases, and 65 - 75% were observed for GIAs. The gene cluster of tobramycin biosynthesis was isolated from the genomic library of Streptomyces tenebrarius using DOI synthase as a probe. Sequencing of 33.9 kb revealed 24 putative open reading frames including the tobramycin biosynthetic gene cluster (13.8 kb) and a transport protein. This cluster encodes proteins homologous to 2-deoxystreptamine biosynthetic enzymes, glycosyltransferase and other aminocyclitols biosynthetic enzymes. Sequence analysis revealed the evolution of DOI synthases from 3-dehydroquinate (DHQ) synthases in actinomycetes. DOI synthases and GIA are therefore useful for cloning biosynthetic genes of DOS-containing aminocyclitol antibiotics or for screening such metabolites producers.
机译：使用基于2-脱氧鞘氨醇（DOI）合酶和L-谷氨酰胺：鞘氨醇氨基转移酶（GIA）核心序列的PCR引物，从氨基糖苷生产商Micromonospora和Streptomyces spp。分离与2-脱氧链胺胺（DOS）生物合成基因同源的基因。 ）。对于DOI合酶，观察到40-45％，对于GIA，观察到65-75％。以DOI合酶为探针，从黄链霉菌基因组文库中分离了妥布霉素生物合成基因簇。 33.9 kb的测序揭示了24个推定的开放阅读框，包括妥布霉素生物合成基因簇（13.8 kb）和转运蛋白。该簇编码与2-脱氧链胺生物合成酶，糖基转移酶和其他氨基环糖醇生物合成酶同源的蛋白质。序列分析揭示了放线菌中DOI合酶从3-脱氢奎宁（DHQ）合酶的进化。因此，DOI合成酶和GIA可用于克隆含DOS的氨基环糖醇抗生素的生物合成基因或用于筛选此类代谢物的产生者。

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