Submerged fermentation of Streptomyces uncialis providing a biotechnology platform for uncialamycin biosynthesis, engineering, and production.
Submerged fermentation of Streptomyces uncialis providing a biotechnology platform for uncialamycin biosynthesis, engineering, and production.
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DOI：
10.1093/jimb/kuab025
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发表时间：
2021-06-04
影响因子：
3.4
通讯作者：
Shen B
中科院分区：
文献类型：
--
作者：
Hindra;Yang D;Luo J;Huang T;Yan X;Adhikari A;Teijaro CN;Ge H;Shen B
uncalamycin （UCM）属于蒽醌融合的10元烯二炔天然产物亚家族，对广泛的人类癌细胞系表现出非凡的细胞毒性。利用UCM的合成类似物作为有效载荷的抗体-药物偶联物正处于临床前开发阶段。UCM仅由unucalis Streptomyces DCA2648在低滴度（~ 0.019 mg/l）的固体琼脂培养基上产生，这限制了其微生物发酵的供应，并阻碍了其在体内途径操作的生物合成和工程研究。在这里，我们报告了培养条件，使UCM生物合成的遗传操作在体内，并允许UCM生产，提高滴度，通过工程的乌氏球菌菌株的深层发酵。具体来说，UCM的滴度提高了近58倍，达到1.1 mg/l，这是通过删除野生型的编码不相关代谢物的生物合成基因簇，化学诱变和途径特异性调节因子的操作来产生工程的unalis菌株，最后对后者进行培养基优化以生产UCM。UCM生物合成的遗传操作被证明是通过灭活工程菌株中选定的基因，其中一个突变菌株积累了天霉素B，天霉素B是一种常见的生物合成中间体，已知为蒽醌融合的enediyne天然产物亚家族。这些发现突出了UCM生物合成、工程和生产的生物技术平台，应该促进其基础研究和转化应用。
Uncialamycin (UCM) belongs to the anthraquinone-fused subfamily of 10-membered enediyne natural products that exhibits an extraordinary cytotoxicity against a wide spectrum of human cancer cell lines. Antibody-drug conjugates, utilizing synthetic analogues of UCM as payloads, are in preclinical development. UCM is exclusively produced by Streptomyces uncialis DCA2648 on solid agar medium with low titers (∼0.019 mg/l), limiting its supply by microbial fermentation and hampering its biosynthetic and engineering studies by in vivo pathway manipulation. Here, we report cultivation conditions that enable genetic manipulation of UCM biosynthesis in vivo and allow UCM production, with improved titers, by submerged fermentation of the engineered S. uncialis strains. Specifically, the titer of UCM was improved nearly 58-fold to ∼1.1 mg/l through the combination of deletion of biosynthetic gene clusters encoding unrelated metabolites from the S. uncialis wild-type, chemical mutagenesis and manipulation of pathway-specific regulators to generate the engineered S. uncialis strains, and finally medium optimization of the latter for UCM production. Genetic manipulation of UCM biosynthesis was demonstrated by inactivating selected genes in the engineered S. uncialis strains, one of which afforded a mutant strain accumulating tiancimycin B, a common biosynthetic intermediate known for the anthraquinone-fused subfamily of enediyne natural products. These findings highlight a biotechnology platform for UCM biosynthesis, engineering, and production that should facilitate both its fundamental studies and translational applications.
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影响因子：
2.7
作者：
Poudel, Yam B.;Rao, Chetana;Gangwar, Sanjeev
通讯作者：
Gangwar, Sanjeev
DOI：
10.1007/s10295-005-0076-x
发表时间：
2006-07-01
影响因子：
3.4
作者：
Dernain, Arnold L.
通讯作者：
Dernain, Arnold L.
DOI：
10.1038/ja.2010.55
发表时间：
2010-08
期刊：
The Journal of antibiotics
影响因子：
--
作者：
通讯作者：
--
影响因子：
11.5
作者：
Lamb, Yvette N.
通讯作者：
Lamb, Yvette N.
影响因子：
3.2
作者：
Hindra;Pak, Patricia;Elliot, Marie A.
通讯作者：
Elliot, Marie A.