2025-12-11 マックス・プランク研究所
High-throughput devices can dramatically accelerate research. Here, 96 samples are tested at once for the enzymatic conversion of formate to formaldehyde—recognizable by the yellow color change.© MPI f. Terrestrial Microbiology/ Franka Eiche
<関連情報>
- https://www.mpg.de/25866898/raw-materials-from-co2
- https://pubs.acs.org/doi/10.1021/acscatal.5c04079
- https://www.sciencedirect.com/science/article/pii/S1096717625001600?via%3Dihub
ギ酸還元酵素のエンジニアリング Engineering a Formic Acid Reductase
Philipp Wichmann,Amelia Cox-Fermandois,Andreas M. Küffner,Uwe Linne,Tobias J. Erb,and Maren Nattermann
ACS Catalysis Published: November 26, 2025
DOI:https://doi.org/10.1021/acscatal.5c04079
Abstract
The formate bioeconomy envisions production of formic acid from CO2 via (electro-)chemical conversion, followed by conversion to the product by engineered microbes or cell-free systems. One prominent way of expanding formate valorization is its reduction to formaldehyde, making highly efficient assimilation cascades accessible. This thermodynamically challenging reaction can be catalyzed by ATP-dependent activation followed by NAD(P)H-dependent reduction. Existing solutions rely on two-step cascades, or fusion enzymes thereof, and are limited by the fast hydrolysis of their formylated intermediates. Here, we show that carboxylic acid reductase can be engineered toward formate reduction, resulting in a single-enzyme solution that does not release intermediates. In addition, we discovered that this enzyme tolerates high formate concentrations when used in Escherichia coli whole-cell conversion, conditions that strongly inhibit existing formate reduction cascades. We therefore provide a valuable addition to the toolbox of synthetic formate reduction, providing an enzyme compatible with applications amenable to high formate titers, such as whole-cell bioconversion or electrobiochemical cascades.
大腸菌におけるホルミルリン酸経路を介したギ酸同化の進化支援工学 Evolution-assisted engineering of formate assimilation via the formyl phosphate route in Escherichia coli
Jenny Bakker, Maximilian Boinot, Karin Schann, Jörg Kahnt, Timo Glatter, Tobias J. Erb, Maren Nattermann, Sebastian Wenk
Metabolic Engineering Available online: 15 October 2025
DOI:https://doi.org/10.1016/j.ymben.2025.10.004
Highlights
- In vivo establishment of the formyl phosphate route for formate reduction to formaldehyde.
- Growth-coupled selection links formate reduction to Thr/Met biosynthesis.
- Adaptive laboratory evolution increases ACK levels, enabling robust formate-dependent growth.
- Evolved E. coli grow with 20–100 mM formate alongside a glucose co-feed.
- Evolution-guided retro-engineering creates strains with balanced ACK/FPR expression and stable formate-dependent growth.
Abstract
The transition towards a sustainable bioeconomy requires the use of alternative feedstocks, with CO2-derived formate emerging as a promising candidate for industrial biotechnology. Despite its beneficial characteristics as a feedstock, microbial assimilation of formate is limited by the inefficiency of naturally evolved formate-fixing pathways. To overcome this limitation, synthetic formate reduction cascades could enable formate assimilation via formaldehyde, a key intermediate of several existing one carbon assimilation pathways. Recently, the formyl phosphate route, combining ATP-dependent activation of formate to formyl phosphate, followed by its reduction to formaldehyde, was developed through enzyme engineering and characterized in vitro. In this work, we successfully established the formyl phosphate route in vivo by developing a selection strategy that couples formate reduction to growth in a threonine/methionine auxotrophic Escherichia coli. Through adaptive laboratory evolution, we achieved formate-dependent growth via this novel pathway. Evolved strains were capable of growing robustly with formate concentrations between 20 mM and 100 mM with glucose in the co-feed. Genomic and proteomic analyses together with activity assays uncovered that formate activation was limiting in vivo. This discovery guided the rational engineering of a strain capable of efficient formate assimilation through the formyl phosphate route. By demonstrating that novel enzyme activities can link formate reduction to cell growth, our study shows how synthetic metabolic routes can be functionally established inside the cell, paving the way for the engineering of more complex synthetic pathways.
