酶促反应结晶研究进展

doi:10.11949/0438-1157.20221223

摘要/Abstract

摘要：

在“双碳”背景下，绿色可持续的酶促反应正受到工业界的广泛关注，但在实际应用中仍面临着诸多挑战，如反应平衡的限制、不稳定产物的分解、酶的产物抑制等。结晶作为一种高效成熟的分离技术，可通过移除液相产物的方式有效解决上述问题。同时，结晶也是晶体产品的“生成”过程，其与酶促反应耦合可一步实现晶体产品的高效、绿色、可控制备。综述了近年来酶促反应结晶的研究进展，介绍了原位产物结晶（ISPC）技术的发展历程，并讨论了结晶与酶促反应耦合时的相互影响关系；从结晶方式和过程控制角度阐述了酶促反应结晶的实现形式和连续化过程；最后，对酶促反应结晶这一耦合过程的发展和应用进行了总结和展望。

关键词: 酶促反应结晶, 原位产物移除, 结晶过程调控

Abstract:

Under the background of“carbon peaking and carbon neutrality”, green and sustainable enzymatic reaction is receiving extensive attention. However, it still faces many challenges in practical application, such as the limitation of reaction equilibrium, the decomposition of unstable products, and the product inhibition of enzyme. As an efficient and mature separation technology, crystallization can effectively solve the above problems by removing liquid phase products. Moreover, crystallization is also the “generation”process of crystals, and the combination of crystallization and enzymatic reaction can realize the efficient, green and controllable preparation of crystal products in one step. This paper reviews the research progress of enzymatic reactive crystallization in recent years. Specifically, the development of in situ product crystallization (ISPC) was introduced, and the interplay between crystallization and enzymatic reaction was discussed. From the perspective of crystallization mode and process control, the realization form and continuous process of enzymatic reactive crystallization were described. Finally, the development and application of this coupled process were summarized and prospected.

Key words: enzymatic reactive crystallization, in situ product removal, crystallization process control

中图分类号:

TQ 026.5

苏伟怡, 丁佳慧, 李春利, 王洪海, 姜艳军. 酶促反应结晶研究进展[J]. 化工学报, 2023, 74(2): 617-629.

Weiyi SU, Jiahui DING, Chunli LI, Honghai WANG, Yanjun JIANG. Research progress of enzymatic reactive crystallization[J]. CIESC Journal, 2023, 74(2): 617-629.

图/表 9

表1 酶促反应结晶实现形式和调控手段汇总

Table 1 Summary of the realization and regulation of enzymatic reaction crystallization

酶	目标产物	结晶方式	调控参数	产率	文献
转氨酶	1-苯基乙胺	加成盐剂		75%	[18]
	1-苯基乙胺	加成盐剂	pH、温度	91.5%	[19]
	(S)-1-(3-甲氧基苯基)乙胺	加成盐剂	晶种		[20]
	(S)-1-(3-甲氧基苯基)乙胺	加成盐剂	晶种		[21]
	3-(2-萘)- l -丙氨酸	自发结晶	温度	93%	[22]
	L-邻苯丙氨酸	自发结晶	pH		[23]
青霉素G酰化酶	氨苄西林	自发结晶		93%	[24]
		自发结晶		97%	[25]
		自发结晶	pH	96%	[26]
		加入晶种	晶种		[27]
		自发结晶	晶种、pH		[28]
		自发结晶	pH	98%	[29]
	阿莫西林	自发结晶	pH	98%	[30]
	头孢氨苄	加络合剂	温度、pH		[31]
	头孢克洛	加络合剂	温度、pH	80%	[32]
	头孢拉定	加络合剂			[33]
葡萄糖氧化酶	葡萄糖酸钙	自发结晶	温度、pH		[34]
氨基酸消旋酶	苏氨酸	自发结晶			[35]
芳香酸脱羧酶	2,6-二羟基苯甲酸	加成盐剂		97%	[36]
嗜热菌蛋白酶	Z-阿斯巴甜	自发结晶	pH	88%	[37]
延胡索酸酶	L-苹果酸钙	自发结晶	温度		[38]

图1 转氨酶催化反应通式

Fig.1 General formula of the transaminase catalyzed reaction

图2 酶促反应结晶用于转氨酶催化的反应，生成一种难溶的1-苯基乙胺-3DPPA盐[18]

Fig.2 In situ product crystallization (ISPC) was combined with the reaction catalyzed by aminotransferase to form insoluble 1-phenylethylamine salt[18]

图3 转氨酶催化连续反应过程示意图[包含供体盐在饱和器的溶解（saturator，左）与产物盐在结晶器的结晶（crystallizer，右）][20]

Fig.3 Schematic diagram of a transaminases-catalyzed continuous reaction process [containing the dissolution of donor salts (left) and crystallization of the product salts (right)][20]

图4 PGA催化合成氨苄西林

Fig.4 Enzymatic synthesis of ampicillin by PGA

图5 过饱和体系构建示意图（实线表示6-APA溶解度，点3对应过饱和溶液中6-APA的浓度）[48]

Fig.5 The schematic pathway for the creation of supersaturated solution(solid line is the thermodynamic solubility of 6-APA, and point 3 corresponds to 6-APA concentration in a supersaturated solution)[48]

图6 泰勒涡旋流反应器示意图[62]

Fig.6 Sketch map of a Taylor-vortex flow reactor[62]

图7 头孢克洛酶促反应结晶（络合结晶）流程图（左为复合反应器，右为酶反应器）[32]

Fig.7 Flow chart of cefaclor enzymatic reactive crystallization (complexation crystallization) (left is the composite reactor, and right is the enzyme reactor)[32]

图8 生物反应结晶器流程图[38]1—生物反应器;2—结晶器;3—缓冲罐;4—泵;5—计时器;6—电磁阀

Fig.8 Flow diagram of the bioreactor-crystallizer[38]1—bioreactor; 2—crystallizer; 3—buffer tank; 4—pump; 5—timer; 6—magnetic valve

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