Identification and Characterization of the Anti-Methicillin-Resistant Staphylococcus aureus WAP-8294A2 Biosynthetic Gene Cluster from Lysobacter enzymogenes OH11
Identification and Characterization of the Anti-Methicillin-Resistant Staphylococcus aureus WAP-8294A2 Biosynthetic Gene Cluster from Lysobacter enzymogenes OH11
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DOI：
10.1128/aac.05370-11
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发表时间：
2011-12-01
影响因子：
4.9
通讯作者：
Du, Liangcheng
中科院分区：
文献类型：
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作者：
Zhang, Wei;Li, Yaoyao;Du, Liangcheng
产酶溶杆菌菌株 OH11 是一种新兴的真菌和细菌性疾病生物防治剂。我们最近完成了它的基因组序列，发现它包含大量推测负责非核糖体肽和聚酮化合物生物合成的基因簇，包括先前鉴定的抗真菌二氢嗜麦芽素（HSAF）。其中一个基因簇包含两个巨大的开放阅读框，共同编码 12 个非核糖体肽合成酶 (NRPS) 模块。其中一种 NRPS 的基因破坏导致野生型中产生的代谢物消失并消除其抗菌活性。一些侧翼基因的破坏也影响了代谢物和抗菌活性。我们随后分离了这种代谢物并对其进行光谱分析。质谱和核磁共振数据显示，其化学结构与WAP-8294A2相同，WAP-8294A2是一种环状脂缩肽，具有有效的抗甲氧西林耐药金黄色葡萄球菌（MRSA）活性，目前正在进行I/II期临床试验。 WAP-8294A2 生物合成基因以前未曾描述过。迄今为止，革兰氏阳性链霉菌一直是抗感染药物的主要来源。溶杆菌是革兰氏阴性土壤/水细菌，可通过基因改造，但尚未得到充分利用。 WAP-8294A2 合成酶代表最大的 NRPS 复合物之一，由 45 个功能域组成。这些基因的鉴定为WAP-8294A2生物合成机制的研究奠定了基础，并为通过生物合成工程在这种新的溶杆菌来源中生产新型抗MRSA抗生素打开了大门。
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.