Naturally-occurring environmental microorganisms may provide ‘green’ andeffective biocontrol tools for disease management in agricultural crops. Due tothe constant threat of resistant pathogens there is a pressing a...Naturally-occurring environmental microorganisms may provide ‘green’ andeffective biocontrol tools for disease management in agricultural crops. Due tothe constant threat of resistant pathogens there is a pressing and continualneed to search for new biocontrol tools. This study investigated the productionof new analogs of WAP-8294A compounds by the biocontrol agent Lysobacterenzymogenes OH11 through biosynthetic engineering. WAP-8294As are afamily of natural cyclic lipodepsipeptides with potent activity against Grampositive bacteria. A series of genetic manipulations was therefore conductedon the accessory genes in the WAP biosynthetic gene cluster. The resultingstrains containing a single-point mutation in ORF4, which was predicted toencode a 2-ketoglutarate dependent dioxygenase, produced deoxy-WAP8294As. This result provides evidence for the function of ORF4 in catalyzing βhydroxylation of the D-asparagine residue in WAP-8294As. In addition, six newanalogs of WAP-8294As were identified by UHPLC-HR-MS/MS. This is the firstattempt to produce new WAP-8294As in Lysobacter and shows that thespectrum of the biocontrol compounds may be expanded through themanipulation of biosynthetic genes.展开更多
文摘Naturally-occurring environmental microorganisms may provide ‘green’ andeffective biocontrol tools for disease management in agricultural crops. Due tothe constant threat of resistant pathogens there is a pressing and continualneed to search for new biocontrol tools. This study investigated the productionof new analogs of WAP-8294A compounds by the biocontrol agent Lysobacterenzymogenes OH11 through biosynthetic engineering. WAP-8294As are afamily of natural cyclic lipodepsipeptides with potent activity against Grampositive bacteria. A series of genetic manipulations was therefore conductedon the accessory genes in the WAP biosynthetic gene cluster. The resultingstrains containing a single-point mutation in ORF4, which was predicted toencode a 2-ketoglutarate dependent dioxygenase, produced deoxy-WAP8294As. This result provides evidence for the function of ORF4 in catalyzing βhydroxylation of the D-asparagine residue in WAP-8294As. In addition, six newanalogs of WAP-8294As were identified by UHPLC-HR-MS/MS. This is the firstattempt to produce new WAP-8294As in Lysobacter and shows that thespectrum of the biocontrol compounds may be expanded through themanipulation of biosynthetic genes.
