基于cMOFs的固定化脂肪酶微反应器的构筑及其扁桃酸催化应用

doi:10.11949/0438-1157.20240698

摘要/Abstract

摘要：

通过高温碳化制备不同系列碳化金属有机骨架（carbonized metal-organic frameworks，cMOFs），应用于制备洋葱伯克霍尔德菌脂解酶（BCL）微反应器，研究了孔结构和孔道微环境对BCL固定化性能的影响，探索了这些微反应器催化拆分扁桃酸对映体反应的性能。结果表明：cMOFs中的cMIL-53（Al）材料对BCL固定性能最佳，且在循环使用九次后，仍保持（R）-扁桃酸转化率（C）为50%，产物的对映体过剩量（ee_p）超过99%，酶选择因子（E）大于200。通过一系列表征发现，经高温碳化处理的cMIL-53（Al）孔间结构增大，有利于BCL进入cMOF微孔结构，增加BCL空间活性位点；同时煅烧使其表面羧酸官能团含量增加，有利于BCL通过氢键与其表面进行吸附固定，最终明显提高拆分催化性能。相较于传统的有机碱催化方法，本研究提出的酶固定化微反应器具有可重复使用、催化时间短、产物易于纯化分离以及环境污染较低等优点，为新型酶固定功能材料构建提供了新的策略。

关键词: 金属有机骨架材料, 碳化, 复合材料, 动力学, 微反应器, 固定化酶

Abstract:

Different series of carbonized metal-organic frameworks (cMOFs) were prepared through high-temperature carbonization and applied for the fabrication of microreactors with Burkholderia cepacia lipase (BCL). The effects of pore structure and pore microenvironment on the immobilization performance of BCL were studied, and the performance of these microreactors in catalytic resolution of mandelic acid enantiomers was explored. The results indicated that the cMIL-53(Al) material from cMOFs exhibited the best immobilization performance for BCL, maintaining a (R)-mandelic acid conversion rate of 50% and an enantiomeric excess (ee_p) value over 99% in nine consecutive catalytic cycles, with an enzyme selectivity (E) value greater than 200. The high-temperature carbonized cMIL-53(Al) showed increased inter-pore structure, allowing BCL to effectively enter the cMOF structure and increase the spatial activity sites of BCL. Additionally, its surface is rich in carboxyl functional groups, enabling BCL to adsorb and immobilize through hydrogen bonding, significantly enhancing the catalytic efficiency. Compared to traditional organic base catalysis methods, the enzyme-immobilized microreactors proposed in this study have the advantages of reusability, short catalytic time, ease of product purification and separation, and lower environmental pollution. This provides a new strategy for the construction of novel enzyme-immobilized functional materials.

Key words: metal-organic framework, carbonization, composites, kinetics, microreactor, immobilized enzyme

中图分类号:

TB 333

李远华, 凌思棋, 封科军, 冯颖, 郭于菁, 谢世桓. 基于cMOFs的固定化脂肪酶微反应器的构筑及其扁桃酸催化应用[J]. 化工学报, 2025, 76(3): 1170-1179.

Yuanhua LI, Siqi LING, Kejun FENG, Ying FENG, Yuching KUO, Shihhuan HSIEH. Construction and catalytic application of immobilized lipase microreactors based on cMOFs for the synthesis of mandelic acid[J]. CIESC Journal, 2025, 76(3): 1170-1179.

图/表 12

图1 BCL催化拆分扁桃酸对映体反应示意图

Fig.1 Resolution of (R,S)-mandelic acid catalyzed by BLC enzyme

图2 四种MOFs的XRD、SEM-EDS和FTIR表征

Fig.2 Charactrization of MIL-53(Al), Al 1,4-NDC, MIL- 100(Al) and MIL-100(Cr) by XRD, SEM and FTIR

图3 四种cMOFs的Raman和SEM图

Fig.3 The Raman and SEM images of cMIL-53(Al), cAl- 1,4-NDC, cMIL-100(Al) and cMIL-100(Cr)

表1 反应温度对BCL催化扁桃酸的影响

Table 1 The effect of temperature on catalyzing mandelic acid by BCL

反应温度/℃	C/%	ee_p/%	ee_s/%	E
25	38.7	>99	74.2	>200
50	50.0	>99	>99	>200

表2 BCL@MIL-53（Al）中BCL浓度对催化拆分扁桃酸对映体反应的影响

Table 2 The effect of the amount of BCL on catalyzing mandelic acid by BCL@MIL-53（Al）

BCL/mg	C/%	ee_p/%	ee_s/%	E
2	15.7	>99	21.4	>200
5	32.0	>99	54.2	>200
10	50.0	>99	>99	>200
15	40.3	>99	67.3	>200

图4 BCL固定前后MIL-53(Al)的XRD、FTIR图和孔径分布

Fig.4 The characterization of XRD, FTIR and pore width of MIL-53(Al) and BCL@MIL-53(Al)

表3 不同反应时间对BCL@MIL-53（Al）催化效率影响

Table 3 The effect of reaction time on catalyzing mandelic acid by BCL@MIL-53（Al）

反应时间/h	C/%	ee_p/%	ee_s/%	E
0.5	50.0	>99	>99	>200
2	50.0	>99	>99	>200
5	50.0	>99	>99	>200

表4 不同反应时间下BCL@MIL-53（Al）重复使用的转化率

Table 4 The effect of reaction time on catalyzing cycles of mandelic acid by BCL@MIL-53（Al）

循环次数	C/%
循环次数	0.5 h	2 h	5 h
1	50.0	50.0	50.0
2	50.0	46.1	42.5
3	50.0	41.6	38.1
4	50.0	32.0	28.4
5	50.0	28.1	21.3

图5 8种BCL微反应器对扁桃酸循环催化的转化率

Fig.5 The conversion yield of (R)-mandelic acid by eight BCL microreactors within nine catalytic cycles

表5 MOFs和cMOFs孔径性质、BCL负载量和羧酸含量

Table 5 The properties of pore structure， loading efficiency of BCL and the amount of COOH group in MOFs and cMOFs

材料	比表面积/ (m²/g)	孔径/nm	BCL 负载量/%	COOH含量/(mmol/g)
MIL-53(Al)	1333.8	0.9	14.5	0.57
cMIL-53(Al)	195.7	4~37	24.5	2.60
Al-1,4-NDC	472.1	0.58	10.6	0.38
cAl-1,4-NDC	292.9	3.9~37	22.3	2.30
MIL-100(Al)	1481.9	1.0~2.2	17.4	0.050
cMIL-100(Al)	17.9	8~37	13.3	2.20
MIL-100(Cr)	2249.6	2.0~5.1	17.6	0.033
cMIL-100(Cr)	759.8	3.4~39	17.8	1.90

图6 四种cMOFs对BCL固定前后的孔径分布对比

Fig. 6 The pore size distribution of four cMOFs immobilized with and without BCL

表6 不同酶对扁桃酸催化性能比较

Table 6 The comparison of catalytical efficiency of mandelic acid in different lipase sources

脂肪酶种类	反应条件	C/%	ee_p/%	E	循环次数	文献
游离脂肪酶	游离的BCL与乙酸乙烯酯溶液在25°C下反应5 h	38.7	99	>200	—	[28]
番木瓜脂肪酶	扁桃酸和NH₃以1∶8摩尔比在25℃下DIPE中反应12 h	49	99	>200	—	[24]
Stutzeri假单胞菌LC2-8脂肪酶	扁桃酸和乙酸乙烯酯以1∶8摩尔比在30℃下DIPE中反应15 h	49.15	99			[17]
Novozym 435	扁桃酸和乙醇以1∶2摩尔比在40℃下1,2-二氯乙烷中反应24 h	30	92	35.2		[18]
双歧伯克霍尔德菌YCJ01	扁桃酸和乙酸乙烯酯以1∶7摩尔比在50℃下DIPE中反应45 min	50	99			[19]
洋葱伯克霍尔德菌脂肪酶@cMIL-53(Al)	扁桃酸和乙酸乙烯酯以1∶5质量比在50℃下DIPE中反应30 min	50	99	>200	9	本文

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