Liu Ziwei, Wang Wei, Zhang Chenghua, Zhang Shiqing, Ma Lijuan, Su Hao, Sheng Xiang
College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, P. R. China.
State Key Laboratory of Engineering Biology for Low-Carbon Manufacturing, Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, P. R. China.
Chemphyschem. 2025 Jul 18;26(14):e202500224. doi: 10.1002/cphc.202500224. Epub 2025 May 21.
2,5-Furandicarboxylic acid decarboxylase (HmfF) belongs to the UbiD family, which employs the cofactor prenylated flavin mononucleotide (prFMN) for catalysis. This enzyme catalyzes the reversible decarboxylation of 2,5-furandicarboxylic acid (FDCA) to produce 2-furancarboxylic acid (F2C). In the present study, quantum chemical calculations are employed to investigate the substrate binding mode and reaction mechanism of HmfF. The calculations demonstrate that HmfF follows a nucleophilic attack mechanism, rather than the 1,3-dipolar cycloaddition mechanism, which is believed more commonly adopted by the prFMN-dependent decarboxylases. Interestingly, the five-membered heterocyclic intermediate characteristic of 1,3-dipolar cycloaddition can also be located. However, it is only a fleeting intermediate that does not contribute to the catalysis. In the proposed mechanism, the reaction initiates with a single CC bond formation between FDCA and prFMN. Then, the CC bond between the carboxylate and the furan group of FDCA breaks to release , followed by a proton transfer from Glu259 to the decarboxylated intermediate, and the subsequent CC bond cleavage to generate the F2C product. Additionally, the infeasibility of HmfF in promoting the decarboxylation of F2C is evaluated computationally, and the obtained information is helpful in designing mutations to enable this reactivity.
