双金属-双酶-辅因子共固定化催化剂用于连续流化学-酶级联催化的酮生成与转化并行反应

doi:10.1016/S1872-2067(26)65145-5

催化学报 ›› 2026, Vol. 88: 457-467.DOI: 10.1016/S1872-2067(26)65145-5

双金属-双酶-辅因子共固定化催化剂用于连续流化学-酶级联催化的酮生成与转化并行反应

刘鹏博^a, 周丽亚^a, 刘鑫龙^a, 付科胜^a, 郭忠旭^a, 原泉^a, 杨恒权^b^,^*(), 刘运亭^a^,^*(), 姜艳军^a^,^*()

^a 河北工业大学化工学院, 天津 300401
^b 山西大学化工学院, 山西太原030006

收稿日期:2026-03-04 接受日期:2026-04-20 出版日期:2026-09-18 发布日期:2026-09-05
通讯作者: *电子信箱: hqyang@sxu.edu.cn (杨恒权),
ytliu@hebut.edu.cn (刘运亭),
yanjunjiang@hebut.edu.cn (姜艳军).
基金资助:
国家重点研发计划(2023YFA0914500);国家自然科学基金(22378096);国家自然科学基金(22578095);河北省自然科学基金(B2023202014)

Bimetallic-bienzyme-cofactor Co-immobilized catalyst for continuous-flow concurrent chemoenzymatic ketone formation-conversion cascade

Pengbo Liu^a, Liya Zhou^a, Xinlong Liu^a, Kesheng Fu^a, Zhongxu Guo^a, Quan Yuan^a, Hengquan Yang^b^,^*(), Yunting Liu^a^,^*(), Yanjun Jiang^a^,^*()

^a School of Chemical Engineering and Technology, Hebei University of Technology, Tianjin 300401, China
^b School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006, Shanxi, China

Received:2026-03-04 Accepted:2026-04-20 Online:2026-09-18 Published:2026-09-05
Supported by:
The National Key Research and Development Program of China(2023YFA0914500);The National Natural Science Foundation of China(22378096);The National Natural Science Foundation of China(22578095);The Natural Science Foundation of Hebei province(B2023202014)

摘要/Abstract

摘要：

酮类化合物广泛存在于天然产物、药物及农用化学品中, 也是构建手性分子的关键前手性中间体, 其生成与转化在有机合成中具有重要意义. 金属催化与酶催化作为两类互补的催化体系, 虽分别在酮生成与转化方面取得显著进展, 但前者通常需苛刻条件, 后者则在酮生成方面应用有限. 将二者耦合的化学-酶级联反应为不对称合成提供了有效途径, 但仍面临反应条件不匹配、一锅体系中相互失活、催化剂难以回收及辅因子成本高等问题. 共固定化策略与连续流技术的结合为提升体系相容性、稳定性及过程强化提供了新思路. 因此, 利用共固定化与连续流技术, 构建高兼容性化学-酶级联催化的酮生成与转化并行反应体系是本文的主要研究思路.
本文将胺脱氢酶与葡萄糖脱氢酶(JaAmDH&GDH)以及辅因子(NADH)共固定于季铵配体修饰的苯基桥联介孔有机硅(PMON^)纳米载体上, 随后包覆聚多巴胺(PDA)壳层, 并在其表面负载PdFe双金属纳米合金, 成功设计了一种双金属-双酶-辅因子共固定催化剂. 其中, PDA壳层不但对内部的酶起到保护作用, 还为外部双金属纳米合金提供负载位点, 实现金属与酶的空间隔离, 有效避免了二者之间的接触失活. PdFe纳米合金在催化芳基碘化物与烯丙醇的Heck偶联反应(酮生成)中, 能以99%的产率获得苄基丙酮类化合物, 重复使用16次后仍保持97%的产率, 展现出优异的催化活性和重复使用性. 密度泛函理论计算表明, PdFe纳米合金的优异活性源于其可降低底物吸附能与关键基元步骤(氧化加成、烯烃插入)的能垒, 并促进催化剂再生. 季铵官能团所带的正电荷使得带负电的酶和辅因子能够通过静电相互作用共固定于载体表面及孔隙中, 构建了辅酶自给自足型生物催化体系. 该体系能够通过不对称还原胺化将酮中间体立体选择性转化为手性γ-芳基胺(酮转化). 与游离酶体系相比, 共固定化体系使辅因子的有效利用水平提高14.6倍, 固定化酶的热稳定性和pH稳定性均显著提高, 重复使用7次仍保持约70%的初始活性. 将该共固定化催化剂应用于连续流体系中, 通过化学-酶级联催化的酮生成与转化并行反应, 实现了手性γ-芳基胺高效合成. 其时空收率高达27.1 g L^-1 h^-1, 较间歇体系提高了21.7倍, 连续运行108 h后, 催化能力保持约87%, 固定化辅因子仍保留约60%以上, 展现出优良的连续生产能力.
综上, 本工作通过共固定化与连续流技术的协同应用, 有效解决了化学-酶级联催化体系中模块兼容性差、级联催化效率低的共性难题, 为构建高效、高兼容性的化学-酶级联催化体系以及高附加值手性化合物的绿色合成提供了新的技术路径与理论依据.

关键词: 化学-酶级联催化, 连续流, 有机硅纳米花, 共固定化, 手性胺

Abstract:

Chemoenzymatic ketone formation-conversion cascades represent a powerful strategy in asymmetric synthesis, yet their efficiency is often limited by catalytic incompatibility and poor operational sustainability. Here, we develop a continuous-flow concurrent chemoenzymatic cascade that integrates a metal-catalyzed Heck coupling of aryl iodides with allylic alcohols (for ketone formation) and an enzymatic asymmetric reductive amination of the resulting ketones, enabling efficient one-pot access to chiral γ-arylamines. The cascade is powered by a bimetallic-bienzyme-cofactor co-immobilized catalyst, which is constructed by spatially incorporating palladium-iron nanoalloy catalysts and a cofactor-self-sufficient biocatalytic system consisting of an amine dehydrogenase, a glucose dehydrogenase and phosphorylated NADH on polydopamine-coated, quaternary-ammonium-functionalized mesoporous organosilica nanoflowers. Implementing this integrated catalyst in a continuous-flow system offers a cost-effective and sustainable platform for asymmetric amine synthesis, delivering markedly enhanced overall catalytic efficiency, high space-time yield, and excellent operational stability.

Key words: Chemoenzymatic cascade, Continuous flow, Organosilica nanoflowers, Co-immobilization, Chiral amines

刘鹏博, 周丽亚, 刘鑫龙, 付科胜, 郭忠旭, 原泉, 杨恒权, 刘运亭, 姜艳军. 双金属-双酶-辅因子共固定化催化剂用于连续流化学-酶级联催化的酮生成与转化并行反应[J]. 催化学报, 2026, 88: 457-467.

Pengbo Liu, Liya Zhou, Xinlong Liu, Kesheng Fu, Zhongxu Guo, Quan Yuan, Hengquan Yang, Yunting Liu, Yanjun Jiang. Bimetallic-bienzyme-cofactor Co-immobilized catalyst for continuous-flow concurrent chemoenzymatic ketone formation-conversion cascade[J]. Chinese Journal of Catalysis, 2026, 88: 457-467.

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链接本文: https://www.cjcatal.com/CN/10.1016/S1872-2067(26)65145-5

https://www.cjcatal.com/CN/Y2026/V88/I9/457

图/表 8

Scheme 1. Sequential (a-c) and concurrent (d,e) chemoenzymatic ketone formation-conversion cascades for the synthesis of chiral alcohols and amines.

Fig. 1. (a) Schematic illustration of the preparation of the bimetal-bienzyme-cofactor co-immobilized catalyst. Representative structural and compositional characterization of M2@PDA@E2C@PMON? at x = 0.3: SEM image (b), TEM image (c), HAADF-STEM image (d), and corresponding elemental mapping for C (e), N (f), Si (g), Pd (h), and Fe (i). (j) High-resolution TEM image of M2@PDA@E2C@PMON?. Compositional and chemical state analysis of the catalysts with varying Fe/Pd ratios (x = 0.2, 0.3, 0.5, 1.0, 2.0, and 3.0): (k) XRD patterns; XPS spectra of the Pd 3d (l) and Fe 2p (m) regions.

Fig. 2. (a) The catalytic performance of PdMx@PDA@E2C@PMON catalysts with different bimetallic ratios (PdMx) in Heck coupling reaction. (b) Reusability of PdFe0.3@PDA@E2C@PMON (0.3 mol% with respect to Pd) in Heck coupling reaction. Reaction conditions: [1a] = 200 mmol/L, [but-3-en-2-ol] = 300 mmol/L, [TEA] = 300 mmol/L, iPrOH/H2O (v:v, 1:1, 4 mL), 80 °C, 12 h. Yields were determined by GC.

Fig. 3. (a) Potential energy profile for the Heck coupling of 1a by PdFe0.3 (111) and Pd (111). (b) Potential energy profile for the product desorption. (c) Structures of the adsorption states and the intermediates in the Heck coupling of 1a by PdFe0.3 (111) and Pd (111).

Fig. 4. (a) Immobilized content of cofactors on PMON? under different conditions: (I) Immobilization of phosphorylated cofactors on PMON? (the initial amount of NADH added is 25-150 μmol). (II) Residual NADH to PMON? after washing treatments with 50 mmol/L PBS buffer at pH 8. (III) Residual NADH to PMON? after washing treatments with 2 mol/L NH4Cl/NH4OH buffer at pH 10.5. The pH-stability (b) and the thermal-stability (c) of M2@PDA@E2C@PMON? compared with free CalAmDH. (d) The yield of 2-Amino-4-phenylbutane (3a) using M2@PDA@E2C@PMON? with different NADH loadings. (e) The yield of AmDH-catalyzed ARA as a function of time. (f) Reusability of M2@PDA@E2C@PMON?. Reaction conditions: [2a] = 100 mmol/L, [glucose] = 150 mmol/L, [M2@PDA@E2C@PMON?] = 25 mg, and NH4Cl/NH4OH buffer (2 mol/L, pH = 10.5)/DMSO (v/v, 2:1, 3 mL).

Table 2 Cofactor catalytic availability of soluble and immobilized CalAmDH with free (248 µmol/L in bulk) and immobilized NADH (36 µmol/gsupport = 248 µmol/L in bulk).

Enzyme form	Cofactor form	Specific activity (U/mg)	NADH available (μmol/L)	NADH available (%)
Soluble	soluble	1.32^a	248	100
Soluble	immobilized	0.17^b	17.5^c	7.1
Immobilized	soluble	0.84^a	248	100
Immobilized	immobilized	0.86^b	257.4^c	103.8

Scheme 2. The substrate scope of the chemoenzymatic enantioselective synthesis for chiral γ-arylamines. A feed solution containing the substrate (100 mmol/L), but-3-en-2-ol (150 mmol/L) and TEA (250 mmol/L) in NH4Cl/NH4OH buffer (2 mol/L, pH 10.5)/DMSO (v/v, 2:1) was pumped into the PBR (immersed in a 50 °C water bath) at a flow rate of 0.2 mL/min.

Fig. 5. (a) The remaining content of immobilized cofactor in the flow system as a function of time. (b) The operational stability of the flow system in the continuous synthesis of 3a.

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双金属-双酶-辅因子共固定化催化剂用于连续流化学-酶级联催化的酮生成与转化并行反应

Bimetallic-bienzyme-cofactor Co-immobilized catalyst for continuous-flow concurrent chemoenzymatic ketone formation-conversion cascade

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