Long-chain N-acyltyrosine synthases from environmental DNA.
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.
Brady
Sean F
SF
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Chao
Carol J
CJ
Clardy
Jon
J
D000215:Acylation; D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D003001:Cloning, Molecular; D003360:Cosmids; D004247:DNA / Q000032:analysis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D015723:Gene Library; D008190:Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009606:Nitrosomonas / Q000378:metabolism; D012987:Soil / Q000032:analysis*; D012988:Soil Microbiology; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis*
D000900:Anti-Bacterial Agents; D012987:Soil; D014443:Tyrosine; D004247:DNA; D008190:Lyases