Metabolic engineering of Escherichia coli for optimized biosynthesis of nicotinamide mononucleotide, a noncanonical redox cofactor
Metabolic engineering of Escherichia coli for optimized biosynthesis of nicotinamide mononucleotide, a noncanonical redox cofactor
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DOI：
10.1186/s12934-020-01415-z
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发表时间：
2020-07-27
影响因子：
6.4
通讯作者：
Li, Han
中科院分区：
文献类型：
--
作者：
Black, William B.;Aspacio, Derek;Li, Han
背景非规范氧化还原辅因子正在成为无细胞生物合成中的重要工具，以增加经济可行性，实现精细控制，并扩大可获得的化学物质的范围。然而，这些非典型的氧化还原辅因子需要生物合成，以实现充分整合与可再生biomanufacturing processing.Results在这项工作中，我们engineeredEscherichia colicells生物合成的非典型的辅因子烟酰胺单核苷酸（NMN+），这已被有效地用于无细胞生物合成。首先，我们开发了一个基于生长的筛选平台来鉴定E中有效的NMN（+）生物合成途径。杆菌其次，我们探索了各种途径的组合和宿主基因的破坏，以达到细胞内水平类似于1.5 mM的NMN+，比细胞的基础水平增加了130倍，在最好的菌株中，其特征在于以前未表征的烟酰胺磷酸核糖基转移酶（NadV）从青枯雷尔氏菌。结论这些结果进一步加深了对NMN（+）在大肠杆菌中有效表达和整合的理解。杆菌这可以实现NMN+导向的生物催化，而不需要外源辅因子供应。
Background Noncanonical redox cofactors are emerging as important tools in cell-free biosynthesis to increase the economic viability, to enable exquisite control, and to expand the range of chemistries accessible. However, these noncanonical redox cofactors need to be biologically synthesized to achieve full integration with renewable biomanufacturing processes.Results In this work, we engineeredEscherichia colicells to biosynthesize the noncanonical cofactor nicotinamide mononucleotide (NMN+), which has been efficiently used in cell-free biosynthesis. First, we developed a growth-based screening platform to identify effective NMN(+)biosynthetic pathways inE. coli. Second, we explored various pathway combinations and host gene disruption to achieve an intracellular level of similar to 1.5 mM NMN+, a 130-fold increase over the cell's basal level, in the best strain, which features a previously uncharacterized nicotinamide phosphoribosyltransferase (NadV) fromRalstonia solanacearum.Last, we revealed mechanisms through which NMN(+)accumulation impactsE. colicell fitness, which sheds light on future work aiming to improve the production of this noncanonical redox cofactor.Conclusion These results further the understanding of effective production and integration of NMN(+)intoE. coli. This may enable the implementation of NMN+-directed biocatalysis without the need for exogenous cofactor supply.