Identification and Characterization of the Anti-Methicillin-Resistant Staphylococcus aureus WAP-8294A2 Biosynthetic Gene Cluster from Lysobacter enzymogenes OH11

Authors

Wei Zhang, University of Nebraska - Lincoln
Yaoyao Li, Shandong University
Guoliang Qian, Nanjing Agricultural UniversityFollow
Yan Wang, University of Nebraska - Lincoln
Haotong Chen, University of Nebraska - Lincoln
Yue-Zhong Li, Shandong University
Fengquan Liu, Nanjing Agricultural University
Yuemao Shen, Shandong UniversityFollow
Liangcheng Du, University of Nebraska - LincolnFollow

Document Type

Article

Date of this Version

2011

Citation

ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, Dec. 2011, p. 5581–5589

Comments

Copyright © 2011.

doi:10.1128/AAC.05370-11

Abstract

Lysobactor enzymogenes strain OH11 is an emerging biological control agent of fungal and bacterial diseases. We recently completed its genome sequence and found it contains a large number of gene clusters putatively responsible for the biosynthesis of nonribosomal peptides and polyketides, including the previously identified antifungal dihydromaltophilin (HSAF). One of the gene clusters contains two huge open reading frames, together encoding 12 modules of nonribosomal peptide synthetases (NRPS). Gene disruption of one of the NRPS led to the disappearance of a metabolite produced in the wild type and the elimination of its antibacterial activity. The metabolite and antibacterial activity were also affected by the disruption of some of the flanking genes. We subsequently isolated this metabolite and subjected it to spectroscopic analysis. The mass spectrometry and nuclear magnetic resonance data showed that its chemical structure is identical to WAP-8294A2, a cyclic lipodepsipeptide with potent antimethicillin-resistant Staphylococcus aureus (MRSA) activity and currently in phase I/II clinical trials. The WAP- 8294A2 biosynthetic genes had not been described previously. So far, the Gram-positive Streptomyces have been the primary source of anti-infectives. Lysobacter are Gram-negative soil/water bacteria that are genetically amendable and have not been well exploited. The WAP-8294A2 synthetase represents one of the largest NRPS complexes, consisting of 45 functional domains. The identification of these genes sets the foundation for the study of the WAP-8294A2 biosynthetic mechanism and opens the door for producing new anti-MRSA antibiotics through biosynthetic engineering in this new source of Lysobacter.