Identification of the biosynthetic gene cluster of A-500359s in Streptomyces griseus SANK60196
Identification of the biosynthetic gene cluster of A-500359s in Streptomyces griseus SANK60196
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DOI：
10.1038/ja.2009.38
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发表时间：
2009-06-01
影响因子：
3.3
通讯作者：
Van Lanen, Steven G.
中科院分区：
文献类型：
--
作者：
Funabashi, Masanori;Nonaka, Koichi;Van Lanen, Steven G.
A-500359是由灰色链霉菌SANK 60196产生的细菌磷酸-N-乙酰胞壁酰-五肽转位酶抑制剂。它们由三个不同的部分组成：5 '-氨基甲酰基尿苷、不饱和己糖醛酸和氨基己内酰胺。鉴定了两个连续的互补序列，覆盖65-kb的DNA区域，编码38个开放阅读框（ORF），参与A-500359的生物合成。逆转录PCR结果显示，38个ORF中的大多数在A-500359 s产生过程中高度表达，但不产生A-500359 s的突变体不表达这些相同的ORF。此外，编码推定的氨基糖苷3 '-磷酸转移酶的orf 21在大肠杆菌和白色链霉菌中异源表达，产生对A-500359 B具有选择性抗性的菌株，表明ORF 21磷酸化不饱和己糖醛酸作为对A-500359 s的自身抗性的机制。总之，数据表明克隆的区域参与A-500359 s的抗性、调节和生物合成。抗生素杂志（2009）62，325-332; doi：10.1038/ja.2009.38; 2009年5月29日在线发表
A-500359s, produced by Streptomyces griseus SANK60196, are inhibitors of bacterial phospho-N-acetylmuramyl-pentapeptide translocase. They are composed of three distinct moieties: a 5'-carbamoyl uridine, an unsaturated hexuronic acid and an aminocaprolactam. Two contiguous cosmids covering a 65-kb region of DNA and encoding 38 open reading frames (ORFs) putatively involved in the biosynthesis of A-500359s were identified. Reverse transcriptase PCR showed that most of the 38 ORFs are highly expressed during A-500359s production, but mutants that do not produce A-500359s did not express these same ORFs. Furthermore, orf21, encoding a putative aminoglycoside 3'-phosphotransferase, was heterologously expressed in Escherichia coli and Streptomyces albus, yielding strains having selective resistance against A-500359B, suggesting that ORF21 phosphorylates the unsaturated hexuronic acid as a mechanism of self-resistance to A-500359s. In total, the data suggest that the cloned region is involved in the resistance, regulation and biosynthesis of A-500359s. The Journal of Antibiotics (2009) 62, 325-332; doi: 10.1038/ja.2009.38; published online 29 May 2009