酶催化制备吡咯啉-5-羧酸和哌啶-6-羧酸的工艺  

Preparation of pyrroline-5-carboxylate and piperidine-6-carboxylate by enzyme catalysis

作　　者：沈忱 黄爱敏 顾正华 辛瑜 张梁 Chen SHEN;Aimin HUANG;Zhenghua GU;Yu XIN;Liang ZHANG(National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi,Jiangsu 214122,China)

机构地区：[1]江南大学粮食发酵工艺与技术国家工程实验室,江苏无锡214122

出　　处：《过程工程学报》2022年第11期1593-1600,共8页The Chinese Journal of Process Engineering

基　　金：江苏省自然科学基金资助项目(编号:BK20160053)。

摘　　要：吡咯啉-5-羧酸(P5C)、哌啶-6-羧酸(P6C)是一类同代谢反应和疾病诊疗密切相关的环形亚胺化合物,在机体内氨基酸代谢中发挥重要作用。本研究利用Thermomicrobium roseum肌氨酸氧化酶对-C-N-键的催化能力建立了一种酶法制备工艺并进行了条件优化。通过薄层层析(TLC)、液质联用(LC-MS)和~1H-NMR核磁共振鉴别出化合物结构,并建立了含量测定方法。单因素实验分析反应温度、pH和底物浓度对产量的影响,并通过响应面得到P5C最佳反应条件:温度为79.33℃、pH为8.54、底物浓度为112.82 mmol/L,P6C最佳反应条件:温度80.19℃、pH为8.31、底物浓度83.58 mmol/L。在此条件下反应30 min,P5C和P6C的产量分别为340.15±9.12, 450.87±8.54 mg/L。反应48 h后,转化率分别达到81.94%和99.99%。研究结果为酶法制备P5C和P6C奠定了理论基础,为吡咯啉、哌啶类化合物的合成提供了新的思路。1-Pyrroline-5-carboxylate(P5C) and 1-piperidine-6-carboxylate(P6C) are cyclic imine compounds closely related to metabolic reactions and disease diagnosis and treatment. They are respectively involved in the metabolism of L-proline and Llysine and play an essential role in maintaining the balance of cell oxidation, regulating the concentration of NADP(H)/NADP,and inducing cell apoptosis. Therefore, it is necessary to develop a green, efficient and sustainable preparation method.Biocatalysis is widely used in various industries such as medicine, chemicals, energy, and materials for its advantages of environment friendliness, economy, mildness, and specificity. In this work, an effective enzymatic method for the preparation of P5C and P6C is established and optimized based on the catalytic ability of the sarcosine oxidase(SOX) against the-C-N-bond. TrSOX is derived from Thermomicrobium roseum, which possesses excellent stability and tolerance. The target compounds are separated and identified by thin-layer chromatography(TLC),liquid chromatography-mass spectrometry(LC-MS), and~1H-NMR, and the concentration determination method was created. The single-factor experiment is conducted to analyze the influence of reaction temperature, pH, and substrate concentration on the product yield, and it is discovered that the optimum reaction temperature, pH and substrate concentration was respective 80℃, 9.5, and 100 mmol/L. Furthermore, reliable models were obtained through response surface design by taking the product yield as the response value. It is found that the best reaction conditions of P5C are 79.33℃, pH=8.54, substrate concentration 112.82 mmol/L, and that of P6C are 80.19℃, pH=8.31, and substrate concentration 83.58 mmol/L. After a 30 min reaction under the above conditions, the yield of P5C and P6C is 340.15±9.12 and 450.87±8.54 mg/L, respectively, which is 1.35% and 1.47% different from the theoretical value. Finally, after 48 h of reaction, the conversion rate of L-proline and L-homoproline reached81.94%

关 键 词：吡咯啉-5-羧酸 哌啶-6-羧酸 薄层层析 产物分析 工艺优化 

分 类 号：O626[理学—有机化学]