Advances on Molecular Modifications of Formate Dehydrogenase for CO₂ Reduction

doi:10.13560/j.cnki.biotech.bull.1985.2024-1038

Abstract

Abstract:

With the urgent global demand for sustainable energy transformation and greenhouse gas emission reduction, the efficient green conversion of CO₂ has become a research hotspot in various fields such as energy, environmental science, and chemical engineering. Especially in the context of combating climate change, CO₂ capture and utilization is seen as one of the key strategies to achieve carbon neutrality. Formate dehydrogenase (FDH) as an important biocatalyst for reducing CO₂ to formate, has shown significant potential in green chemistry and bioenergy conversion. However, it still has certain limitations regarding catalytic activity, thermal stability, and coenzyme specificity. In recent years, with the continuous development of protein engineering technology and molecular biology, researchers have proposed a variety of strategies to modify the performance of FDH, significantly enhancing its application prospects. For example, enzyme engineering means such as directed mutation and structure optimization can improve the substrate affinity of the enzyme, enhance the rigidity and conformational stability of the enzyme, and change the coenzyme binding site to broaden its application prospects in the process of green transformation. In this paper, the research progress of FDH catalytic CO₂ reduction in recent years will be comprehensively summarized, focusing on the improvement measures of FDH in terms of catalytic efficiency, thermal stability, coenzyme specificity, etc., the specific strategies adopted in the molecular transformation process will be discussed, and the rules of these strategies will be summarized, in order to provide new ideas and methods for the molecular transformation of FDH in the future and promote the development of its application in CO₂ reduction reaction. In addition, with the development of artificial intelligence, machine learning, gene editing and other technologies, the molecular modification of FDH will be more efficient and precise in the future, and these emerging technologies are expected to screen out FDH mutants with excellent performance in a short time, providing a feasible green solution in solving the global climate change and energy crisis.

Key words: CO? reduction, formate dehydrogenase, molecular modification, catalytic efficiency, thermal stability, cofactor specificity, formate, NAD?/NADH

LU Feng, HUANG Yu-hong, LIN Yan-na, MA Fu-qiang. Advances on Molecular Modifications of Formate Dehydrogenase for CO₂ Reduction[J]. Biotechnology Bulletin, 2025, 41(3): 14-24.

Figures/Tables 6

Fig. 1 Flowchart of multi-enzyme cascade catalysis for CO2 to methanol synthesisFDH: Formate dehydrogenase; FaldDH: formaldehyde dehydrogenase; ADH: alcohol dehydrogenase; NADH: reduced nicotinamide adenine dinucleotide; NAD+: oxidized nicotinamide adenine dinucleotide; W/Mo: tungsten/molybdenum

Table 1 Types and kinetic parameters of FDH for CO2 reduction

类型 Types	物种 Species	FDH	分子量 Mw /kD	k_cat /（s^-1）	K_M /（mmol·L^-1）	k_cat/K_M / （mmol·L^-1·s^-1）	参考文献 Reference
NAD⁺依赖型FDH	Candida boidinii	CbFDH	41.0	0.015 0	31.3	0.000 400	［20］
	Thiobacillus sp. KNK65MA	TsFDH	45.0	0.318	9.23	0.034 0	［20］
	Thermochaetoides thermophila DSM 1495	CtFDH	45.0	0.023 0	0.320	0.069 0	［21］
	Candida methylica	CmFDH	42.0	0.008 00	0.780	0.010 0	［21］
	Thermothelomyces thermophilus ATCC 42464	MtFDH	42.0	0.100	0.400	0.250	［22］
	Paracoccus sp. MKU1	PsFDH	44.0	0.073 0	0.928	0.079 0	［23］
金属依赖型FDH	Desulfovibrio desulfuricans	DdFDH	135	46.6	0.015 7	2.97×10³	［24］
	Desulfovibrio vulgaris Hildenborough	DvFdhAB	97.4	315	0.420	750	［25］
	Cupriavidus necator	FdsABG	178	11.0	2.70	4.07	［26］
	Acetobacterium woodii	FdhF1/2	169	372	3.80	97.9	［27］
	Escherichia coli	EcFDH-H	79.0	1.00	8.30	0.120	［28］
	Clostridium Ijungdahlii	ClFDH	80.0	0.012 0	7.27	0.001 65	［29］
	Clostridium ljungdahlii	ClFDH	75.0	5.66	66.2	0.085 5	［30］
	Clostridium autoethanogenum	CaFDH	75.0	4.00	23.2	0.170	［31］
	Clostridium coskatii	CcFDH	75.0	5.62	59.7	0.094 0	［31］
	Clostridium ragsdalei	CrFDH	75.0	3.28	31.2	0.110	［31］
	Clostridium carboxidivorans P7T	FDH_H_CloCa	80.7	0.080 0	0.050 0	1.60	［32］
	Pseudomonas oxalaticus	Formate Dehydrogenase	315	3.00	40.0	0.075 0	［33-34］
	Desulfosporosinus acididurans	DaFDH	93.0	4.09	34.9	0.117	［18］
	Paraclostridium bifermentans	PbFDH	93.0	4.45	30.6	0.145	［18］
	Clostridium scatologenes	CsFDH	79.0	1.55	48.9	0.031 7	［18］

Fig. 2 Structure and catalytic mechanism of CbFDHA: Three-dimensional structure of CbFDH (PDB: 6D4C). The two dimers are colored in red and green, while the NAD+ cofactor is colored in orange. B: Active center of CbFDH and amino acid residues closed to NAD+. C: Mechanism of action of NAD+-dependent FDH for formate oxidation (green arrows) or CO2 reduction (inverse sense, pink arrows)

Fig. 3 Three-dimensional structure and cofactor diagram of E. coli FDH NA: Three-dimensional structure of Escherichia coli FDH N (PDB: 1KQF). The three polypeptide chains are indicated by blue (chain A), pink (chain B) and green (chain C). The molybdopterin (in chain A), the five [4Fe-4S] clusters (in chain A and chain B) and the two heme (chain C) are displayed in pink, green and red, respectively. B: Graphic of the molybdenum cofactor and its chemical formula structure in E. coli FDH N

Fig. 4 Feasible mechanism of CO2 reduction to formic acid with metal-dependent FDHA: CO2 is electrostatically captured by positively charged arginine residues of FDH. B: Hydrogen mercaptan bound to metal Mo/W attacks CO2 captured by arginine residues, and the valence state is oxidized from four to six. C: Under the action of accepting proton electrons and releasing formic acid, the valence state of metal Mo/W is reduced from six to four, and the initial state is restored

Fig. 5 Schematic diagram of catalytic activity, thermal stability and coenzyme preference of FDH enhanced by molecular modification

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