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.

中文翻译：

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使用 ThDP 依赖酶从糠醛和甲醛生物合成头孢呋辛中间体 2-呋喃基羟甲基酮

( Z )-2-methoxyimino-2-(furyl-2-yl) acetic acid ammonium salt (SMIA) 是合成头孢呋辛的关键中间体。目前SMIA的常规化学合成存在一些缺点，例如成本高和环境污染。在这项研究中，我们提出了一种从 2-呋喃基羟甲基酮 (2-FHMK) 制备 SMIA 的替代路线，其中使用生物催化剂代替以糠醛和甲醛为原料的传统化学合成 2-乙酰呋喃。硫胺素二磷酸 (ThDP) 依赖性酶最初是在大肠杆菌中筛选和组装的。来自硫酸杆菌的丙酮酸脱羧酶sp。hq2 (SsPDC) 被确定为筛选的 21 种酶中的最佳酶。此外，优化了 2-FHMK 的酶催化合成，并使用 MBP 标签来增加其溶解度并提高 SsPDC 的催化效率。最后，63 g L ⁻¹ 2-FHMK 在 1 L 反应器中产生（96.2% 产率，5.25 g L ⁻¹ h ⁻¹时空产率）。该路线为工业规模的低成本和绿色生产SMIA提供了一种有前途的替代方案，并为甲醛和糠醛的再利用提供了另一种策略。