World Journal of Microbiology and Biotechnology ( IF 4.0 ) Pub Date : 2023-10-21 , DOI: 10.1007/s11274-023-03739-5 Hong-Ling Shi 1, 2 , Shu-Wei Yuan 3 , Xiao-Qi Xi 2 , Yu-Li Xie 2 , Chao Yue 2 , Ying-Jun Zhang 4 , Lun-Guang Yao 2 , Chuang Xue 1 , Cun-Duo Tang 2
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Formate dehydrogenase (FDH) is a D-2-hydroxy acid dehydrogenase, which can reversibly reduce CO2 to formate and thus act as non-photosynthetic CO2 reductase. In order to increase catalytic efficiency of formate dehydrogenase for CO2 reduction, two mutants V328I/F285W and V354G/F285W were obtained of which reduction activity was about two times more than the parent CbFDHM2, and the formate production from CO2 catalyzed by mutants were 2.9 and 2.7-fold higher than that of the parent CbFDHM2. The mutants had greater potential in CO2 reduction. The optimal temperature for V328I/F285W and V354G/F285W was 55 °C, and they showed increasement of relative activity under 45 °C to 55 °C compared with parent. The optimal pH for the mutants was 9.0, and they showed excellent stability in pH 4.0–11.5. The kcat/Km values of mutants were 1.75 times higher than that of the parent. Then the molecular basis for its improvement of biochemical characteristics were preliminarily elucidated by computer-aided methods. All of these results further established a solid foundation for molecular modification of formate dehydrogenase and CO2 reduction.
Graphical abstract
中文翻译：
用于提高二氧化碳转化为甲酸盐潜力的甲酸脱氢酶工程
甲酸脱氢酶(FDH)是一种D -2-羟基酸脱氢酶，它可以可逆地将CO 2还原为甲酸，从而充当非光合CO 2还原酶。为了提高甲酸脱氢酶对CO 2还原的催化效率，获得了两个突变体V328I/F285W和V354G/F285W，其还原活性约为亲本Cb FDH M2的两倍，并催化CO 2生产甲酸。突变体比亲本Cb FDH M2高 2.9 倍和 2.7 倍。突变体在CO 2还原方面具有更大的潜力。V328I/F285W和V354G/F285W的最适温度为55℃，与亲本相比，它们在45℃至55℃之间表现出相对活性的增加。突变体的最适pH为9.0，并且在pH 4.0-11.5范围内表现出优异的稳定性。突变体的k cat / K m值比亲本高1.75倍。然后通过计算机辅助方法初步阐明了其改善生化特性的分子基础。这些结果进一步为甲酸脱氢酶的分子修饰和CO 2还原奠定了坚实的基础。
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