烯烃还原酶和有机染料协同催化邻二胺的立体选择性生物合成

doi:10.1016/S1872-2067(24)60168-3

催化学报 ›› 2025, Vol. 68: 223-229.DOI: 10.1016/S1872-2067(24)60168-3

烯烃还原酶和有机染料协同催化邻二胺的立体选择性生物合成

石丰铭^a, 陈彬^a, 余进海^a, 朱睿琦^a, 郑煜^b, 黄小强^a^,^*()

^a南京大学, 化学化工学院, 化学和生物医药创新中心（ChemBIC）, 化生医药学科交叉研究中心, 配位化学国家重点实验室, 江苏南京 210023
^b南京林业大学, 林产化学与材料国际创新高地, 江苏省林业资源高效加工利用协同创新中心, 江苏南京 210037

收稿日期:2024-08-30 接受日期:2024-10-16 出版日期:2025-01-18 发布日期:2025-01-02
通讯作者: * 电子信箱: huangx513@nju.edu.cn (黄小强).
基金资助:
国家自然科学基金(22277053);国家重点研发计划(2022YFA0913000);江苏省自然科学基金项目(BK20220760);南京大学卓越计划(ZYJH004)

Enantioselective biosynthesis of vicinal diamines enabled by synergistic photo/biocatalysis consisting of an ene-reductase and a green-light-excited organic dye

Fengming Shi^a, Bin Chen^a, Jinhai Yu^a, Ruiqi Zhu^a, Yu Zheng^b, Xiaoqiang Huang^a^,^*()

^aState Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center (ChemBIC), ChemBioMed Interdisciplinary Research Center, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, Jiangsu, China
^bJiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, International Innovation Center for Forest Chemicals and Materials, Nanjing Forestry University, Nanjing 210037, Jiangsu, China

Received:2024-08-30 Accepted:2024-10-16 Online:2025-01-18 Published:2025-01-02
Contact: * E-mail: huangx513@nju.edu.cn (X. Huang).
Supported by:
National Natural Science Foundation of China(22277053);National Key Research and Development Program of China(2022YFA0913000);Natural Science Foundation of Jiangsu Province(BK20220760);Excellent Research Program of Nanjing University(ZYJH004)

摘要/Abstract

摘要：

邻二胺在制药和材料领域展现出了广阔的应用前景, 因此, 其合成方法吸引了有机化学研究者的广泛关注. 烯胺的氢胺化反应是一种极具潜力的合成邻二胺方法, 尽管目前研究实例相对较少，且主要依赖于铜氢物种和手性磷酸催化, 但其重要性不容忽视. 生物催化因其在选择性、可进化性和可持续性方面具有显著的优势, 为这一领域提供了新的研究视角. 然而, 关于烯胺的生物催化氢胺化反应至今尚未见报道, 这无疑为该领域的研究开辟了新的机遇.
本课题组前期通过光酶催化策略, 成功利用硫胺素依赖的自由基酰基转移酶、烯烃还原酶和亚胺还原酶，实现了多类非天然生物合成转化. 在此基础上, 本文进一步发展了一种由黄素单核苷酸依赖的烯烃还原酶和有机染料罗丹明B组成的协同双催化系统, 该系统能够高效地将烯酰胺转化为邻二胺. 在加入光催化剂罗丹明B后, 反应可在530-540 nm的绿光激发下顺利进行，产生氮中心自由基. 这些自由基随后与预先固定在酶口袋中的烯胺底物发生反应，形成C-N键, 并经过立体选择性的氢原子转移，最终生成手性1,2-二胺产物. 实验结果表明, 无论是给电子基团和还是吸电子基团取代的烯酰胺都能参与反应, 显示出良好的官能团兼容性. 对于吡啶鎓盐类底物, 各种取代基的化合物均能够以较好的收率和对映选择性进行反应. 此外, 该策略成功实现了0.1 mmol规模的放大反应, 且保持了先前的收率和对映选择性. 机理研究进一步证实了反应过程中氮中心自由基和电子供体-受体(EDA)络合物的存在. 我们推测, 反应中罗丹明B被绿光激发后, 通过与EDA络合物间的能量转移或电子转移, 产生自由基并引发后续反应.
综上, 本文开发了一种光酶催化立体选择性氢胺化合成手性邻二胺的新方法, 该方法具有卓越的立体选择性(>99% e.e.), 展示了光酶催化在产生和控制高度活性和短寿命的自由基中间体方面的巨大潜力. 本文利用光引发的自由基物种开发酶的非天然催化功能, 为合成化学中长期存在的一些挑战提供新的解决方案.

关键词: 光酶催化, 邻二胺, 不对称合成, 氢胺化, 绿光激发反应

Abstract:

Vicinal diamines are key motifs widely-found in many pharmaceuticals and biologically active molecules. An appealing approach for synthesizing these molecules is the amination of enamines, but few examples have been explored. With the utilization of nitrogen-centered radicals (NCRs), here we present the development of a dual bio-/photo-catalytic system for achieving enantioselective hydroamination of enamides, which can give easy access to diverse enantioenriched vicinal diamines. These reactions progress efficiently under green light excitation and exhibit excellent enantioselectivities (up to >99% enantiomeric excess). Mechanistic studies uncovered the synergistic effect of the enzyme and the externally added organophotoredox catalyst Rhodamine B (RhB). This work demonstrates the effectiveness of photobiocatalysis to generate and control high-energy radical intermediates, addressing a long-standing challenge in chemical synthesis.

Key words: Photobiocatalysis, Vicinal diamines, Asymmetric synthesis, Hydroamination, Green light excitation

石丰铭, 陈彬, 余进海, 朱睿琦, 郑煜, 黄小强. 烯烃还原酶和有机染料协同催化邻二胺的立体选择性生物合成[J]. 催化学报, 2025, 68: 223-229.

Fengming Shi, Bin Chen, Jinhai Yu, Ruiqi Zhu, Yu Zheng, Xiaoqiang Huang. Enantioselective biosynthesis of vicinal diamines enabled by synergistic photo/biocatalysis consisting of an ene-reductase and a green-light-excited organic dye[J]. Chinese Journal of Catalysis, 2025, 68: 223-229.

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链接本文: https://www.cjcatal.com/CN/10.1016/S1872-2067(24)60168-3

https://www.cjcatal.com/CN/Y2025/V68/I1/223

图/表 4

Fig. 1. Photodriven ERED-enabled modular access to chiral vicinal diamines. (A) Two pathways for amination of enamines. (B) Previously reported photoenzymatic hydroaminations of styrenes. (C) This work: hydroaminations of enamides through a non-natural green-light-tri- ggered N-N cleavage radical mechanism. NCRs, nitrogen-centered radicals; GDH, glucose dehydrogenase; NADP+, the oxidized form of nicotinamide adenine dinucleotide phosphate; Glu, glucose; FMNsq, flavin semiquinone; HAT, hydrogen atom transfer.

Table 1 Development of photoenzymatic hydroamination.

Entry	EREDs	Deviation from the above conditions ^a	Yield ^b (%)	e.e. ^c (%)
1	GluER-M5 ^d	w/o RhB, 420−430 nm LEDs	26	98
2	GluER-M5	none	72	98
3	GluER	none	45	99
4	YersER	none	46	96
5	XenA	none	trace	n.d.
6	OYE1-F296A	none	19	68
7	NostocER	none	65	72
8	CsER	none	42	71
9	GluER-M5	1 mol% enzyme loading	48	98
10	GluER-M5	w/o GDH / NADP⁺/ Glu	n.d.	n.a.
11	none	None	n.d.	n.a.
12	none	5 mol% FMN-Na added	n.d.	n.a.
13	GluER-M5	Dark	trace	n.a.
14	GluER-M5	w/o RhB, 300-320 nm LEDs	8	n.d.

Fig. 2. Scope Investigation. Standard conditions: enamide 1 (0.008 mmol), N-amidopyridinium salt 2 (0.032 mmol), GluER-M5 (2 mol%), GDH (4 U), NADP+ (1.5 mol%), glucose (0.024 mmol), 20% v/v DMSO in KPi buffer (100 mmol L-1, pH = 6.0) were stirred for 16 h at room temperature and illuminated with 530?540 nm LEDs under a N2 atmosphere, total volume of the reaction is 2.0 mL. The yields were determined by LC-MS and reported as the average of duplicates. The e.e. was determined by HPLC analysis on a chiral stationary phase. a Isolated yields were based on 0.10-mmol-scale reactions. b Mutant GluER-Y177F-F269V was used as the enzyme.

Fig. 3. Mechanism studies. (A) Radical trapping experiment with 1,1-diphenylethene. (B) UV-vis absorption spectra. 1a (0.4 mmol L-1), 2a (0.4 mmol L-1), GluER-M5 (0.05 mmol L-1), sodium dithionite (0.15 mmol L-1). (C) Emission spectrum of green LEDs and absorption spectra of key components. RhB (0.01 mmol L-1), GluER-M5 (0.05 mmol L-1), sodium dithionite (0.15 mmol L-1), 2a (0.4 mmol L-1). (D) Stern-Volmer quenching study. RhB (0.01 mM). Samples were irradiated at 530 nm and emission was measured at 582 nm. (E) Proposed catalytic cycle, where the dot curvature represents the chiral enzyme active site. EDA, electron donor-acceptor; ET, energy transfer; FMNox, flavin mononucleotide; FMNhq, flavin hydroquinone.

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烯烃还原酶和有机染料协同催化邻二胺的立体选择性生物合成

Enantioselective biosynthesis of vicinal diamines enabled by synergistic photo/biocatalysis consisting of an ene-reductase and a green-light-excited organic dye

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