Brady Sean F, Chao Carol J, Clardy Jon
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Appl Environ Microbiol. 2004 Nov;70(11):6865-70. doi: 10.1128/AEM.70.11.6865-6870.2004.
The heterologous expression of DNA extracted directly from environmental samples (environmental DNA [eDNA]) in easily cultured hosts provides access to natural products produced by previously inaccessible microorganisms. When eDNA cosmid libraries were screened in Escherichia coli for antibacterially active clones, long-chain N-acyltyrosine-producing clones were found in every eDNA library. These apparently common natural products have not been previously described from screening extracts of cultured bacteria for biologically active natural products. Of the 11 long-chain N-acyl amino acid synthases (NASs) that were characterized, 10 are unique sequences. A predicted protein of previously unknown function from Nitrosomonas europaea, a gram-negative nitrifying beta-proteobacterium, is 14 to 37% identical to eDNA NASs. When cloned into E. coli, this open reading frame confers the production of long-chain N-acyltyrosines to the host and is therefore the first NAS from a cultured bacterium to be functionally characterized. Understanding the role that long-chain N-acyl amino acids play in soil microbial communities should now be feasible with the identification of a cultured organism that has the genetic capacity to produce these compounds.
直接从环境样品中提取的DNA(环境DNA [eDNA])在易于培养的宿主中的异源表达,为获取以前难以获得的微生物产生的天然产物提供了途径。当在大肠杆菌中筛选eDNA粘粒文库以寻找具有抗菌活性的克隆时,在每个eDNA文库中都发现了产生长链N-酰基酪氨酸的克隆。这些明显常见的天然产物以前从未在培养细菌的提取物中筛选生物活性天然产物时被描述过。在已鉴定的11种长链N-酰基氨基酸合成酶(NAS)中,有10种是独特的序列。来自欧洲亚硝化单胞菌(一种革兰氏阴性硝化β-变形菌)的一种功能未知的预测蛋白与eDNA NAS的同源性为14%至37%。当克隆到大肠杆菌中时,这个开放阅读框赋予宿主产生长链N-酰基酪氨酸的能力,因此它是第一个从培养细菌中功能鉴定的NAS。随着一种具有产生这些化合物遗传能力的培养生物的鉴定,现在了解长链N-酰基氨基酸在土壤微生物群落中所起的作用应该是可行的。