Heterologous Production and Yield Improvement of Epothilones in Burkholderiales Strain DSM 7029
Heterologous Production and Yield Improvement of Epothilones in Burkholderiales Strain DSM 7029
复制标题
伯克霍尔德氏菌菌株 DSM 7029 中埃坡霉素的异源生产和产量提高。
DOI：
10.1021/acschembio.7b00097
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发表时间：
2017-07-01
影响因子：
4
通讯作者：
Zhang, Youming
中科院分区：
文献类型：
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作者：
Bian, Xiaoying;Tang, Biao;Zhang, Youming
微生物天然产物生物合成基因簇的克隆及其在合适宿主中的异源表达已被证明是提高有价值天然产物产量和开始挖掘微生物中潜在天然产物的可行途径。粘细菌是一种多产的新型生物活性天然产物的来源，只有有限的选择，已被开发的异源宿主。在这里，我们描述了使用伯克霍尔德菌菌株DSM 7029作为一个潜在的异源宿主的粘细菌次级代谢产物的功能性表达。利用一种新建立的电穿孔方法，将来自纤维堆囊粘杆菌的56 kb埃博霉素生物合成基因簇通过转座导入菌株DSM 7029的染色体中。检测到埃坡霉素A、B、C和D的产生，尽管它们的产率低。培养基的优化，引入外源甲基丙二酰辅酶A生物合成途径，以及稀有tRNA基因的过表达导致埃博霉素的总产量增加约75倍，达到307 μg L-1。这些结果表明，菌株DSM 7029具有产生具有合理滴度的埃博霉素的潜力，并且可能是其他粘细菌聚酮酶和非核糖体肽合成酶的异源表达的广泛适用的宿主，从而加快基因组挖掘的过程。
The cloning of microbial natural product biosynthetic gene clusters and their heterologous expression in a suitable host have proven to be a feasible approach to improve the yield of valuable natural products and to begin mining cryptic natural products in microorganisms. Myxobacteria are a prolific source of novel bioactive natural products with only limited choices of heterologous hosts that have been exploited. Here, we describe the use of Burkholderiales strain DSM 7029 as a potential heterologous host for the functional expression of myxobacterial secondary metabolites. Using a newly established electroporation procedure, the 56 kb epothilone biosynthetic gene cluster from the myxobacterium Sorangium cellulosum was introduced into the chromosome of strain DSM 7029 by transposition. Production of epothilones A, B, C, and D was detected despite their yields being low. Optimization of the medium, introduction of the exogenous methylmalonyl-CoA biosynthetic pathway, and overexpression of rare tRNA genes resulted in an approximately 75-fold increase in the total yields of epothilones to 307 μg L-1. These results show that strain DSM 7029 has the potential to produce epothilones with reasonable titers and might be a broadly applicable host for the heterologous expression of other myxobacterial polyketide synthases and nonribosomal peptide synthetases, expediting the process of genome mining.