Meta-omic characterization of the marine invertebrate microbial consortium that produces the chemotherapeutic natural product ET-743.
Meta-omic characterization of the marine invertebrate microbial consortium that produces the chemotherapeutic natural product ET-743.
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海洋无脊椎动物微生物联盟的荟萃分析，产生化学治疗性天然产物ET-743。
DOI：
10.1021/cb200244t
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发表时间：
2011-11-18
影响因子：
4
通讯作者：
Sherman DH
中科院分区：
文献类型：
--
作者：
Rath CM;Janto B;Earl J;Ahmed A;Hu FZ;Hiller L;Dahlgren M;Kreft R;Yu F;Wolff JJ;Kweon HK;Christiansen MA;Håkansson K;Williams RM;Ehrlich GD;Sherman DH
在许多大型生物体中，由于无法识别和培养产生共生微生物，有效的生物活性天然产物的最终来源仍然难以捉摸。作为开发元组学方法来识别和表征来自无脊椎动物来源的微生物群落的天然产物途径的模型系统，我们选择研究 ET-743 (Yondelis®) 生物合成途径。该分子是一种经批准的抗癌剂，从被囊动物 Ecteinascidia turbinata 中以低丰度 (10−4–10−5% w/w) 获得，并通过漫长的半合成过程产生适量的临床用途。基于与三种细菌次级代谢产物的结构相似性，我们假设 ET-743 是海洋细菌共生体的产物。通过对被囊类动物/微生物群落的总 DNA 进行宏基因组测序，我们靶向并组装了一个包含 25 个基因的 35 kb 重叠群，这些基因构成了这种有价值的抗癌剂的 NRPS 生物合成途径的核心。基于两个大的未连锁重叠群的密码子使用的严格序列分析表明 Candidatus Endoecteinascidia frumentensis 产生 ET-743 代谢物。随后的宏蛋白质组学分析证实了三种关键生物合成蛋白质的表达。此外，体外验证了用于组装 ET-743 四氢异喹啉核心的酶的预测活性。这项工作为通过代谢工程直接生产药物和新类似物奠定了基础。我们期望所描述的跨学科方法适用于多种宿主共生系统，为药物发现和开发产生有价值的天然产物。
In many macroorganisms, the ultimate source of potent biologically active natural products has remained elusive due to an inability to identify and culture the producing symbiotic microorganisms. As a model system for developing a meta-omic approach to identify and characterize natural product pathways from invertebrate-derived microbial consortia we chose to investigate the ET-743 (Yondelis®) biosynthetic pathway. This molecule is an approved anti-cancer agent obtained in low abundance (10−4–10−5% w/w) from the tunicate Ecteinascidia turbinata, and is generated in suitable quantities for clinical use by a lengthy semi-synthetic process. Based on structural similarities to three bacterial secondary metabolites, we hypothesized that ET-743 is the product of a marine bacterial symbiont. Using metagenomic sequencing of total DNA from the tunicate/microbial consortium we targeted and assembled a 35 kb contig containing 25 genes that comprise the core of the NRPS biosynthetic pathway for this valuable anti-cancer agent. Rigorous sequence analysis based on codon usage of two large unlinked contigs suggests that Candidatus Endoecteinascidia frumentensis produces the ET-743 metabolite. Subsequent metaproteomic analysis confirmed expression of three key biosynthetic proteins. Moreover, the predicted activity of an enzyme for assembly of the tetrahydroisoquinoline core of ET-743 was verified in vitro. This work provides a foundation for direct production of the drug and new analogs through metabolic engineering. We expect that the interdisciplinary approach described is applicable to diverse host-symbiont systems that generate valuable natural products for drug discovery and development.
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影响因子：
3.4
作者：
Deutsch, Eric W.;Mendoza, Luis;Shteynberg, David;Farrah, Terry;Lam, Henry;Tasman, Natalie;Sun, Zhi;Nilsson, Erik;Pratt, Brian;Prazen, Bryan;Eng, Jimmy K.;Martin, Daniel B.;Nesvizhskii, Alexey I.;Aebersold, Ruedi
通讯作者：
Aebersold, Ruedi
DOI：
10.1073/pnas.0806305105
发表时间：
2008-09-02
影响因子：
11.1
作者：
Calderone, Christopher T.;Bumpus, Stefanie B.;Magarvey, Nathan A.
通讯作者：
Magarvey, Nathan A.
影响因子：
14.8
作者：
Fisch, Katja M.;Gurgui, Cristian;Piel, Joern
通讯作者：
Piel, Joern
影响因子：
14.8
作者：
Donia, Mohamed S.;Ravel, Jacques;Schmidt, Eric W.
通讯作者：
Schmidt, Eric W.
DOI：
10.1007/bf01965946
发表时间：
1980-01-01
期刊：
EXPERIENTIA
影响因子：
--
作者：
ARAI, T;TAKAHASHI, K;KUBO, A
通讯作者：
KUBO, A