Unnatural biosynthesis by an engineered microorganism with heterologously expressed natural enzymes and an artificial metalloenzyme.
Unnatural biosynthesis by an engineered microorganism with heterologously expressed natural enzymes and an artificial metalloenzyme.
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由工程微生物与异源表达的天然酶和人工金属酶进行的非自然生物合成。
DOI：
10.1038/s41557-021-00801-3
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发表时间：
2021-12
期刊：
影响因子：
21.8
通讯作者：
Hartwig JF
中科院分区：
文献类型：
--
作者：
Huang J;Liu Z;Bloomer BJ;Clark DS;Mukhopadhyay A;Keasling JD;Hartwig JF
合成生物学使微生物宿主能够产生复杂的分子，这些分子是由罕见或难以培养的生物体产生的，但这些分子的结构仅限于由天然酶催化的化学反应形成的分子。将催化非天然反应的人工金属酶（ArMs）整合到代谢网络中可以拓宽微生物合成产生的分子的该高速缓存。我们报告了一个工程微生物细胞表达异源生物合成途径，其中包含天然酶和ArMs，产生高非对映选择性的非天然产物。为了创造这种杂交生物合成生物，我们改造了大肠杆菌（E.大肠杆菌）中的一种，以及一种含有铱-卟啉复合物的ArM，所述ArM用异源转运系统转运到细胞中。我们通过进化ArM和选择合适的基因诱导和培养条件来提高非天然产物的非对映选择性和产物滴度。这项工作表明，合成生物学和合成化学可以在整个细胞中与天然和人工酶一起产生以前无法进入自然界的分子。
Synthetic biology enables microbial hosts to produce complex molecules that are otherwise produced by organisms that are rare or difficult to cultivate, but the structures of these molecules are limited to those formed by chemical reactions catalyzed by natural enzymes. The integration of artificial metalloenzymes (ArMs) that catalyze unnatural reactions into metabolic networks could broaden the cache of molecules produced biosynthetically by microorganisms. We report an engineered microbial cell expressing a heterologous biosynthetic pathway, which contains both natural enzymes and ArMs, that produces an unnatural product with high diastereoselectivity. To create this hybrid biosynthetic organism, we engineered Escherichia coli (E. coli) with a heterologous terpene biosynthetic pathway and an ArM containing an iridium-porphyrin complex that was transported into the cell with a heterologous transport system. We improved the diastereoselectivity and product titer of the unnatural product by evolving the ArM and selecting the appropriate gene induction and cultivation conditions. This work shows that synthetic biology and synthetic chemistry can produce, together with natural and artificial enzymes in whole cells, molecules that were previously inaccessible to nature.
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影响因子：
8.4
作者：
Heinisch T;Schwizer F;Garabedian B;Csibra E;Jeschek M;Vallapurackal J;Pinheiro VB;Marlière P;Panke S;Ward TR
通讯作者：
Ward TR
影响因子：
15
作者：
Dydio, Pawel;Key, Hanna M.;Hartwig, John F.
通讯作者：
Hartwig, John F.
影响因子：
12.9
作者：
Grimm, Alexander R.;Sauer, Daniel F.;Schwaneberg, Ulrich
通讯作者：
Schwaneberg, Ulrich
影响因子：
15
作者：
Lelyveld VS;Brustad E;Arnold FH;Jasanoff A
通讯作者：
Jasanoff A
影响因子：
15
作者：
Skander, M;Humbert, N;Ward, TR
通讯作者：
Ward, TR