卤醇脱卤酶催化合成(R)-1-氯-3-苯氧基-2-丙醇的工艺优化

doi:10.16085/j.issn.1000-6613.2018-2413

摘要/Abstract

摘要：

以卤醇脱卤酶重组湿菌体E. coli BL21（pET28a-HHDH）为催化剂，催化外消旋的1-氯-3-苯氧基-2-丙醇的动力学拆分可以获得光学纯的(R)-1-氯-3-苯氧基-2-丙醇。本文系统地研究了卤醇脱卤酶催化合成光学纯(R)-1-氯-3-苯氧基-2-丙醇的影响因素，对反应pH、反应温度、菌体浓度、亲核试剂 $N 3 -$ 浓度和底物浓度进行了探究。结果表明，卤醇脱卤酶催化合成(R)-1-氯-3-苯氧基-2-丙醇的最佳工艺条件为：pH为7.0，反应温度为28℃，菌体浓度为22.5g/L，亲核试剂NaN₃的浓度为50mmol/L，底物外消旋1-氯-3-苯氧基-2-丙醇的浓度为10mmol/L。在此工艺条件下，(R)-1-氯-3-苯氧基-2-丙醇的ee值和收率分别为100%和16.97%。

关键词: 卤醇脱卤酶, 1-氯-3-苯氧基-2-丙醇, 催化, 工艺

Abstract:

Halohydrin dehalogenase (HHDH) was produced by recombinant strain E. coli BL21 (pET28a-HHDH). This recombinant E. coli was used as whole-cell catalysis to synthesize enantiopure (R)-1-chloro-3-phenoxy-2-propanol. The influencing factors of HHDH catalyzed synthesis of (R)-1-chloro-3-phenoxy-2-propanol were studied in the experiment. The reaction pH, reaction temperature, catalyst concentration, nucleophilic concentration (NaN₃) and substrate concentration were investigated. The results showed that the optimal conditions were as follows: pH was 7.0, reaction temperature was 28℃, the concentration of catalyst was 22.5g/L, the concentration of sodium azide was 50mmol/L, the concentration of racemic 1-chloro-3-phenoxy-2-propanol was 10mmol/L. Under these conditions, the ee and yield of (R)-1-chloro-3-phenoxy-2-propanol was 100% and 16.97%, respectively.

Key words: halohydrin dehalogenase, 1-chloro-3-phenoxy-2-propanol, catalysis, synthesis technique

中图分类号:

TQ032.4

亚香菊,王于齐,朱鑫海,经玉洁,林茜,薛锋,高健. 卤醇脱卤酶催化合成(R)-1-氯-3-苯氧基-2-丙醇的工艺优化[J]. 化工进展, 2019, 38(08): 3823-3828.

Xiangju YA,Yuqi WANG,Xinhai ZHU,Yujie JING,Xi LIN,Feng XUE,Jian GAO. Process optimization for synthesis of (R)-1-chloro-3-phenoxy-2-propanol by halohydrin dehalogenase[J]. Chemical Industry and Engineering Progress, 2019, 38(08): 3823-3828.

图/表 8

表1 不同pH的Tris-SO4缓冲液

pH	加入酸量/mL
pH	0.2mol/L Tris	0.05mol/L H₂SO₄
7.0	50	45.7
7.5	50	40.3
8.0	50	29.2
8.5	50	14.7

图1 卤醇脱卤酶催化从β-氯代醇生成β-叠氮醇的生物催化拆分反应

图2 不同pH对催化拆分的影响

图3 不同温度对催化拆分的影响

图4 菌体浓度对动力学拆分的影响

图5 亲核试剂浓度对催化拆分的影响

图6 底物浓度对催化拆分的影响

图7 卤醇脱卤酶催化合成(R)-1-氯-3-苯氧基-2-丙醇

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