Multiomics Analysis Provides Insight into the Laboratory Evolution of Escherichia coli toward the Metabolic Usage of Fluorinated Indoles.
Multiomics Analysis Provides Insight into the Laboratory Evolution of Escherichia coli toward the Metabolic Usage of Fluorinated Indoles.
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DOI：
10.1021/acscentsci.0c00679
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发表时间：
2021-01-27
影响因子：
18.2
通讯作者：
Koksch B
中科院分区：
文献类型：
--
作者：
Agostini F;Sinn L;Petras D;Schipp CJ;Kubyshkin V;Berger AA;Dorrestein PC;Rappsilber J;Budisa N;Koksch B
众所周知，有机氟化合物对不同生境中的各种生物都有毒性，人类历史上曾利用化学氟化来开发治疗药物或农用杀虫剂。另一方面，到目前为止，一些研究已经证明，在适当的条件下，生命系统(特别是细菌)可以容忍代谢中存在的氟化分子(例如氨基酸类似物)，甚至将它们重新用作合成蛋白质等细胞大分子的替代构件。了解这些现象背后的分子机制将极大地促进重组蛋白质和多肽药物的生物技术合成。然而，关于活细胞长期暴露于含氟氨基酸的代谢影响的信息仍然很少。在此，我们报道了大肠杆菌在人工加氟生境中的长期繁殖，该生境产生了两个自然适应于含氟氨基酸的菌株。特别是，我们施加了选择性压力，迫使色氨酸(Trp)营养缺陷型菌株使用4-或5-氟吲哚作为必要的前体来原位合成Trp类似物，然后将它们整合到细胞蛋白质组中。我们发现，完全适应这两种氟化色氨酸类似物需要少量的基因突变，但伴随着调控网络、膜完整性和蛋白质折叠的质量控制的大量重排。这些发现突出了非天然氨基酸适应背后的细胞机制，并为合成生物学和生物技术中新的微生物菌株的生物工程提供了分子基础。我们消除了大肠杆菌产生典型氨基酸色氨酸的代谢能力，并介绍了以4-和5-氟吲哚为前体在体内合成两个含氟类似物。
Organofluorine compounds are known to be toxic to a broad variety of living beings in different habitats, and chemical fluorination has been historically exploited by mankind for the development of therapeutic drugs or agricultural pesticides. On the other hand, several studies so far have demonstrated that, under appropriate conditions, living systems (in particular bacteria) can tolerate the presence of fluorinated molecules (e.g., amino acids analogues) within their metabolism and even repurpose them as alternative building blocks for the synthesis of cellular macromolecules such as proteins. Understanding the molecular mechanism behind these phenomena would greatly advance approaches to the biotechnological synthesis of recombinant proteins and peptide drugs. However, information about the metabolic effects of long-term exposure of living cells to fluorinated amino acids remains scarce. Hereby, we report the long-term propagation of Escherichia coli (E. coli) in an artificially fluorinated habitat that yielded two strains naturally adapted to live on fluorinated amino acids. In particular, we applied selective pressure to force a tryptophan (Trp)-auxotrophic strain to use either 4- or 5-fluoroindole as essential precursors for the in situ synthesis of Trp analogues, followed by their incorporation in the cellular proteome. We found that full adaptation to both fluorinated Trp analogues requires a low number of genetic mutations but is accompanied by large rearrangements in regulatory networks, membrane integrity, and quality control of protein folding. These findings highlight the cellular mechanisms behind the adaptation to unnatural amino acids and provide the molecular foundation for bioengineering of novel microbial strains for synthetic biology and biotechnology. We eliminated the metabolic ability of E. coli to produce the canonical amino acid tryptophan and introduced synthesis of two fluorinated analogues in vivo using 4- and 5-fluoroindole as precursors.
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DOI：
10.1073/pnas.0805309105
发表时间：
2008-12-09
影响因子：
11.1
作者：
Huang, Kerwyn Casey;Mukhopadhyay, Ranjan;Wingreen, Ned S.
通讯作者：
Wingreen, Ned S.
影响因子：
16.6
作者：
Choe, Donghui;Lee, Jun Hyoung;Cho, Byung-Kwan
通讯作者：
Cho, Byung-Kwan
影响因子：
3.2
作者：
Batchelor, E;Walthers, D;Goulian, M
通讯作者：
Goulian, M
影响因子：
3
作者：
Lu, Na;Zhang, Bin;Liu, Hongyan
通讯作者：
Liu, Hongyan
影响因子：
5.8
作者：
Choi, Yongwook;Chan, Agnes P.
通讯作者：
Chan, Agnes P.