Long-chain N-acyltyrosine synthases from environmental DNA
Long-chain N-acyltyrosine synthases from environmental DNA
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DOI：
10.1128/aem.70.11.6865-6870.2004
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发表时间：
2004-11-01
影响因子：
4.4
通讯作者：
Clardy, J
中科院分区：
文献类型：
--
作者：
Brady, SF;Chao, CJ;Clardy, J
直接从环境样品中提取的DNA（环境DNA [eDNA]）在容易培养的宿主中的异源表达提供了获得由先前不可接近的微生物产生的天然产物的途径。当在大肠杆菌中筛选cDNA粘粒文库中的抗菌活性克隆时，在每个cDNA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没有从筛选培养细菌的提取物中描述过生物活性天然产物。在已表征的11种长链N-酰基氨基酸合酶（NAS）中，有10种是独特序列。一种来自欧洲亚硝化单胞菌（一种革兰氏阴性硝化β-变形杆菌）的先前未知功能的预测蛋白质与eDNA NAS具有14至37%的相同性。克隆到E. coli中，该开放阅读框架赋予宿主长链N-酰基酪氨酸的产生，因此是待功能表征的来自培养细菌的第一个NAS。了解长链N-酰基氨基酸在土壤微生物群落中发挥的作用，现在应该是可行的，具有产生这些化合物的遗传能力的培养生物体的鉴定。
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.