Direct valorization of biomass into value-added chemicals is attractive yet challenging. Herein, we report a one-pot synthesis of 2,5-bis(hydroxymethyl)furan (BHMF) and its diacetate from inexpensive fructose and sugar syrup in the aqueous phase via sequential chemical dehydration, biocatalytic reduction, and esterification. A robust alcohol dehydrogenase from Escherichia coli (EcYjgB) was identified for HMF reduction, with high catalytic efficiencies toward 5-hydroxymethylfurfural (HMF) (k_cat/K_m, 1300 s^–1 mM^–1) as well as NADPH (9700 s^–1 mM^–1). Recombinant E. coli harboring EcYjgB (E. coli_EcYjgB) showed high tolerance toward highly toxic and strongly inhibitory biobased furans, enabling smooth reduction of 1 M HMF and 0.5 M furfural. Its practicality was demonstrated by gram-scale synthesis of BHMF with a 79% isolated yield at approximately 126 g/L substrate loading. Biocatalytic process intensification was realized by improving cell membrane permeability using ethyl acetate, providing up to 15 g/L h BHMF productivity. A concurrent biocatalytic cascade for producing BHMF diacetate from HMF was constructed by combining E. coli_EcYjgB and Mycobacterium smegmatis acyltransferase (MsAcT) variant N94A, affording the desired product with a 93% yield in 1 h. Starting from crude HMF obtained via chemical dehydration and organic solvent evaporation, BHMF and its diacetate were produced with >99 and 88% yields, respectively. Finally, direct synthesis of these furan-based products from sugars was performed by chemobiocatalysis, without isolation of any intermediates, resulting in yields of up to 59% (relative to sugar). This work may pave the way for scalable production of furan-based chemicals from inexpensive renewable biomass.

中文翻译：

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水介质中生物质一锅化学生物催化生产2,5-双(羟甲基)呋喃及其二酯

将生物质直接增值为增值化学品既有吸引力又具有挑战性。在此，我们报道了在水相中通过连续化学脱水、生物催化还原和酯化从廉价果糖和糖浆一锅合成2,5-双（羟甲基）呋喃（BHMF）及其二乙酸酯。一种来自大肠杆菌( Ec YjgB)的强效醇脱氢酶被鉴定用于 HMF 还原，对 5-羟甲基糠醛 (HMF) ( k _cat / K _m , 1300 s ^–1 mM ^–1 ) 以及 NADPH (9700 s ^–1 mM ^–1 ) 具有高催化效率。重组大肠杆菌窝藏Ec YjgB ( E. coli _ Ec YjgB) 对高毒性和强抑制性生物基呋喃表现出高耐受性，能够顺利还原 1 M HMF 和 0.5 M 糠醛。克级合成 BHMF 证明了其实用性，在底物负载量约为 126 g/L 时，分离产率为 79%。通过使用乙酸乙酯改善细胞膜渗透性来实现生物催化过程强化，提供高达 15 g/L h BHMF 生产率。通过结合大肠杆菌_ Ec YjgB和耻垢分枝杆菌酰基转移酶 ( MsAcT) 变体 N94A，在 1 小时内提供所需产物，收率 93%。从通过化学脱水和有机溶剂蒸发获得的粗 HMF 开始，BHMF 及其二乙酸酯的产率分别 >99% 和 88%。最后，通过化学生物催化从糖直接合成这些呋喃基产品，无需分离任何中间体，产率高达 59%（相对于糖）。这项工作可能为利用廉价的可再生生物质大规模生产呋喃类化学品铺平道路。