当前位置：
X-MOL 学术
›
Microb. Biotechnol.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Functional replacement of isoprenoid pathways in Rhodobacter sphaeroides.
Microbial Biotechnology ( IF 4.8 ) Pub Date : 2020-03-24 , DOI: 10.1111/1751-7915.13562 Enrico Orsi 1 , Jules Beekwilder 2 , Dewi van Gelder 1 , Adèle van Houwelingen 2 , Gerrit Eggink 1, 3 , Servé W M Kengen 4 , Ruud A Weusthuis 1
Microbial Biotechnology ( IF 4.8 ) Pub Date : 2020-03-24 , DOI: 10.1111/1751-7915.13562 Enrico Orsi 1 , Jules Beekwilder 2 , Dewi van Gelder 1 , Adèle van Houwelingen 2 , Gerrit Eggink 1, 3 , Servé W M Kengen 4 , Ruud A Weusthuis 1
Affiliation
|
Advances in synthetic biology and metabolic engineering have proven the potential of introducing metabolic by‐passes within cell factories. These pathways can provide a more efficient alternative to endogenous counterparts due to their insensitivity to host's regulatory mechanisms. In this work, we replaced the endogenous essential 2‐C‐methyl‐D‐erythritol 4‐phosphate (MEP) pathway for isoprenoid biosynthesis in the industrially relevant bacterium Rhodobacter sphaeroides by an orthogonal metabolic route. The native 2‐C‐methyl‐D‐erythritol 4‐phosphate (MEP) pathway was successfully replaced by a heterologous mevalonate (MVA) pathway from a related bacterium. The functional replacement was confirmed by analysis of the reporter molecule amorpha‐4,11‐diene after cultivation with [4‐13C]glucose. The engineered R. sphaeroides strain relying exclusively on the MVA pathway was completely functional in conditions for sesquiterpene production and, upon increased expression of the MVA enzymes, it reached even higher sesquiterpene yields than the control strain coexpressing both MEP and MVA modules. This work represents an example where substitution of an essential biochemical pathway by an alternative, heterologous pathway leads to enhanced biosynthetic performance.
中文翻译：
球形红杆菌中类异戊二烯途径的功能替代。
合成生物学和代谢工程的进步已经证明了在细胞工厂内引入代谢旁路的潜力。由于这些途径对宿主的调节机制不敏感，因此可以提供比内源性对应物更有效的替代方案。在这项工作中，我们通过正交代谢途径取代了工业相关细菌球形红杆菌中类异戊二烯生物合成的内源必需2-C-甲基-D-赤藓糖醇4-磷酸（MEP）途径。天然的 2-C-甲基-D-赤藓糖醇 4-磷酸 (MEP) 途径成功地被来自相关细菌的异源甲羟戊酸 (MVA) 途径取代。用 [4- 13 C] 葡萄糖培养后，通过分析报告分子 amorpha-4,11-diene 证实了功能替代。完全依赖于 MVA 途径的工程化R. sphaeroides菌株在倍半萜生产条件下完全发挥功能，并且在 MVA 酶表达增加时，它达到了比共表达 MEP 和 MVA 模块的对照菌株更高的倍半萜产量。这项工作代表了一个例子，其中用替代的异源途径替代重要的生化途径可以增强生物合成性能。
更新日期：2020-03-24
中文翻译：
球形红杆菌中类异戊二烯途径的功能替代。
合成生物学和代谢工程的进步已经证明了在细胞工厂内引入代谢旁路的潜力。由于这些途径对宿主的调节机制不敏感，因此可以提供比内源性对应物更有效的替代方案。在这项工作中，我们通过正交代谢途径取代了工业相关细菌球形红杆菌中类异戊二烯生物合成的内源必需2-C-甲基-D-赤藓糖醇4-磷酸（MEP）途径。天然的 2-C-甲基-D-赤藓糖醇 4-磷酸 (MEP) 途径成功地被来自相关细菌的异源甲羟戊酸 (MVA) 途径取代。用 [4- 13 C] 葡萄糖培养后，通过分析报告分子 amorpha-4,11-diene 证实了功能替代。完全依赖于 MVA 途径的工程化R. sphaeroides菌株在倍半萜生产条件下完全发挥功能，并且在 MVA 酶表达增加时，它达到了比共表达 MEP 和 MVA 模块的对照菌株更高的倍半萜产量。这项工作代表了一个例子，其中用替代的异源途径替代重要的生化途径可以增强生物合成性能。
京公网安备 11010802027423号