一种新颖深海微生物羧酸酯酶制备(R)-苯乙醇的研究

doi:10.11978/2016081

热带海洋学报 ›› 2017, Vol. 36 ›› Issue (3): 55-60.doi: 10.11978/2016081CSTR: 32234.14.2016081

一种新颖深海微生物羧酸酯酶制备(R)-苯乙醇的研究

黄锦龙^1,²(), 张云^1,³, 孙爱君^1,³, 胡云峰^1,^3,⁴()

1. 中国科学院南海海洋研究所, 中国科学院热带海洋生物资源与生态重点实验室, 广东广州 510301
2. 中国科学院大学, 北京 100049
3. 中国科学院南海海洋研究所, 广东省海洋药物重点实验室, 广东广州 510301
4. 南海生物资源开发和利用协同创新中心, 广东广州 510275

收稿日期:2016-08-31 修回日期:2016-09-22 出版日期:2017-05-20 发布日期:2017-06-01
作者简介:
作者简介：黄锦龙(1986—), 男, 广西壮族自治区东兴市人, 博士研究生, 研究方向为生物催化与酶工程。E-mail: jl_huang@163.com
基金资助:
中国科学院战略性先导科技专项(XDA11030404);广州市科技计划项目(201510010012);国家自然科学基金青年科学基金项目(21302199)

Enantioselective production of (R)-1-phenylethanol by a novel marine microbial carboxylesterase

Jinlong HUANG^1,²(), Yun ZHANG^1,³, Aijun SUN^1,³, Yunfeng HU^1,^3,⁴()

1. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, Guangdong 510301, China
2. University of Chinese Academy of Sciences, Beijing 100049, China
3. Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, Guangdong 510301, China
4. South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Guangzhou, Guangdong 510275, China

Received:2016-08-31 Revised:2016-09-22 Online:2017-05-20 Published:2017-06-01
Supported by:
Strategic Priority Research Program of the Chinese Academy of Sciences (XDA11030404);Guangzhou Science and Technology Plan Projects (201510010012);National Natural Science Foundation of China (21302199)

摘要/Abstract

摘要：

深海微生物酯酶BSE00077能够有效地水解拆分消旋乙酸苏合香酯, 制备(R)-1-苯乙醇。优化的反应条件为50mmol•L^-1底物, 10%体积分数的正庚烷作为溶剂, 在pH 7.0和30℃条件下反应2h。经过优化后, 目标产物(R)-1-苯乙醇的对映体过量值超过99%, 转化率超过30%。我们发现酯酶BSE00077也能够拆分不同酰基链长的1-苯乙醇酯, 酰基链长会影响1-苯乙醇的对映体过量值。深海来源的酯酶BSE00077作为生物催化剂在工业上制备(R)-1-苯乙醇和其他手性化工产品具有很好的应用潜力。

关键词: 深海微生物酯酶, (R)-1-苯乙醇, 水解反应, 动力学拆分

Abstract:

Deep-sea microbial esterase BSE00077 can efficiently resolve (±)-1-phenylethyl acetate and generate (R)-1- phenylethanol. The optimal working conditions for the enzymatic resolution of (±)-1-phenylethyl acetate by marine microbial esterase BSE00077 were found to be: 50mmol/L (±)-1-phenylethyl acetate, 10% n-heptane (v/v), pH 7.0, 30°C for 2h. After process optimization, desired optically pure product (R)-1-phenylethanol was obtained with an enantiomeric excess of over 99% and a conversion of over 30%. Esterase BSE00077 can also enzymatically resolve some other racemic 1-phenylethyl esters with different chain lengths. We found that the chain lengths could greatly affect the enantiomeric excess of desired product. Esterase BSE00077 is a novel marine microbial esterase with great potential in the asymmetric synthesis of (R)-1-phenylethanol as well as of other chiral chemicals in industry.

Key words: deep-sea microbial esterase, (R)-1-phenylethanol, hydrolysis, kinetic resolution

黄锦龙, 张云, 孙爱君, 胡云峰. 一种新颖深海微生物羧酸酯酶制备(R)-苯乙醇的研究[J]. 热带海洋学报, 2017, 36(3): 55-60.

Jinlong HUANG, Yun ZHANG, Aijun SUN, Yunfeng HU. Enantioselective production of (R)-1-phenylethanol by a novel marine microbial carboxylesterase[J]. Journal of Tropical Oceanography, 2017, 36(3): 55-60.

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参考文献 24

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温度/℃	产物对映体过量值/%	底物转化率/%	pH	产物对映体过量值/%	底物转化率/%
25	96.17 ± 0.47	37.12 ± 1.32	6	99.50 ± 0	9.20 ± 1.72
30	96.21 ± 0.88	42.51 ± 1.45	7	93.27 ± 1.08	41.64 ± 2.44
35	95.53 ± 0.35	44.54 ± 1.74	8	91.94 ± 0.64	45.32 ± 1.69
40	95.25 ± 0.95	37.28 ± 0.77	9	91.23 ± 0.52	47.10 ± 3.07
45	85.30 ± 0.81	9.96 ± 1.31

有机溶剂	lg P^*	产物对映体过量值/%	底物转化率/%	对映体比值E
对照		94.35 ± 1.05	27.90 ± 0.79	49.38 ± 4.27
二甲基亚砜	-1.35	93.75 ± 0.67	31.76 ± 1.44	47.97 ± 4.38
甲醇	-0.72	93.63 ± 0.45	7.33 ± 1.38	32.80 ± 1.95
甲苯	2.68	>99.00	5.04 ± 0.69	209.65 ± 1.56
二甲苯	3.14	94.14 ± 0.88	14.37 ± 1.05	39.24 ± 3.68
正己烷	3.94	94.49 ± 0.95	31.73 ± 3.41	55.36 ± 6.25
正庚烷	4.47	94.81 ± 0.68	31.72 ± 2.44	58.53 ± 5.12
正辛烷	5.01	94.23 ± 0.79	32.03 ± 1.75	52.77 ± 5.76

时间/h	产物对映体过量值/%	底物转化率/%
0.5	99.19 ± 0.14	23.79 ± 1.03
1	99.12 ± 0.12	25.68 ± 0.92
2	99.03 ± 0.09	30.71 ± 0.67
3	98.14 ± 0.48	36.22 ± 2.31
4	97.00 ± 0.47	40.60 ± 1.94
5	96.45 ± 1.01	42.33 ± 2.04
6	94.48 ± 0.89	43.57 ± 1.87
7	94.53 ± 0.36	43.53 ± 1.44
8	94.43 ± 0.42	43.61 ± 1.37

底物	反应时间/h	起始反应速率/(mmol·L^-1·h^-1)	产物对映体过量值/%	底物转化率/%
乙酸苏合香酯	0.5	23.79	99.19 ± 0.14	23.79 ± 1.03
丙酸苏合香酯	0.5	23.84	90.22 ± 0.51	23.84 ± 1.55
丁酸苏合香酯	0.5	10.40	91.67 ± 0.73	10.40 ± 0.28

酶(来源)	反应方式	R/S- 苯乙醇	对映体过量值/%	参考文献
脂肪酶Novozym 435(商业酶)	转酯化	S	87.5	(严祥辉等, 2009)
C. antarctica 脂肪酶(商业酶)	水解	S	99.5	(Fan et al, 2011)
C. lipolytica 脂肪酶(商业酶)	转酯化	S	99.5	(范卫东等, 2012)
酯酶BSE00077 (本研究)	水解	R	99.0

一种新颖深海微生物羧酸酯酶制备(R)-苯乙醇的研究

Enantioselective production of (R)-1-phenylethanol by a novel marine microbial carboxylesterase

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