An anaerobic bacterium host system for heterologous expression of natural product biosynthetic gene clusters
An anaerobic bacterium host system for heterologous expression of natural product biosynthetic gene clusters
复制标题
用于异源表达天然产物生物合成基因簇的厌氧细菌宿主系统
DOI：
10.1038/s41467-019-11673-0
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发表时间：
2019-08-14
影响因子：
16.6
通讯作者：
Chen, Yihua
中科院分区：
文献类型：
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作者：
Hao, Tingting;Xie, Zhoujie;Chen, Yihua
厌氧细菌是生物和化学多样性的一个被忽视的丰富来源。由于培养的挑战和遗传的棘手性，评估其生物合成基因簇（BGC）的次级代谢产物生产的能力需要一个有效的异源表达系统。然而，这种主机系统仍然不可用。在这里，我们使用兼性厌氧菌变形链球菌UA 159作为异源宿主，用于从厌氧菌表达BGC。建立了一种基于天然感受态的大片段DNA克隆（NabLC）技术，该技术可将DNA片段直接移动到40-kb，并将一个73.7-kb的BGC整合到S. mutansUA 159通过三轮NabLC克隆。使用这个系统，我们确定了一种抗浸润化合物，变环素，从未定义的BGC从人类口腔细菌。我们预期该宿主系统将用于从厌氧细菌异源表达BGC。
Anaerobic bacteria represent an overlooked rich source of biological and chemical diversity. Due to the challenge of cultivation and genetic intractability, assessing the capability of their biosynthetic gene clusters (BGCs) for secondary metabolite production requires an efficient heterologous expression system. However, this kind of host system is still unavailable. Here, we use the facultative anaerobeStreptococcus mutansUA159 as a heterologous host for the expression of BGCs from anaerobic bacteria. A natural competence based large DNA fragment cloning (NabLC) technique was developed, which can move DNA fragments up to 40-kb directly and integrate a 73.7-kb BGC to the genome ofS. mutansUA159 via three rounds of NabLC cloning. Using this system, we identify an anti-infiltration compound, mutanocyclin, from undefined BGCs from human oral bacteria. We anticipate this host system will be useful for heterologous expression of BGCs from anaerobic bacteria.