The (Z)-2-methoxyimino-2-(furyl-2-yl) acetic acid ammonium salt (SMIA) is a key intermediate used for the synthesis of cefuroxime. The current conventional chemical synthesis of SMIA shows some disadvantages, such as high cost and environmental pollution. In this study, we have proposed an alternative route to prepare SMIA from 2-furylhydroxymethylketone (2-FHMK), in which a biocatalyst was used to replace the traditional chemical synthesis of 2-acetylfuran utilizing furfural and formaldehyde as raw materials. Thiamine diphosphate (ThDP)-dependent enzymes were initially screened and assembled in Escherichia coli. Pyruvate decarboxylase from Sulfobacillus sp. hq2 (SsPDC) was identified as the optimal enzyme among the 21 enzymes screened. Furthermore, the enzyme-catalyzed synthesis of 2-FHMK was optimized and an MBP tag was used to increase its solubility and improve the catalytic efficiency of SsPDC. Finally, 63 g L⁻¹ 2-FHMK was produced in a 1-L reactor (96.2% yield, 5.25 g L⁻¹ h⁻¹ space-time yield). This route provides a promising alternative for the low-cost and green production of SMIA on an industrial scale and provides another strategy for the reuse of formaldehyde and furfural.