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

Abstract

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

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.

Figures/Tables 7

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References 24

[1]	范卫东, 范永仙, 陈小龙, 2012. 用Candida lipolytica脂肪酶手性转酯拆分农药中间体rac-1-苯乙醇[J]. 浙江农业科学, (12): 1689-1692.
[2]	严祥辉, 薛屏, 2009. 用壳聚糖修饰的MCM-48固定化脂肪酶拆分(R, S)-1-苯乙醇[J]. 宁夏大学学报(自然科学版), 30(1): 50-54.
	YAN XIANHUI, XUE PING, 2009. Chiral resolution of (R, S)-1-phenylethanol catalyzed by lipase immobilized on MCM-48 coated chitosan[J]. Journal of Ningxia University (Natural Science Edition), 30(1): 50-54 (in Chinses).
[3]	BLASCO M A, GRÖGER H, 2014. Enzymatic resolution of racemates with a ‘remote’ stereogenic center as an efficient tool in drug, flavor and vitamin synthesis[J]. Bioorganic & Medicinal Chemistry, 22(20): 5539-5546.
[4]	BORNSCHEUER U T, 2014. Enzymes in lipid modification: past achievements and current trends[J]. European Journal of Lipid Science and Technology, 116(10): 1322-1331.
[5]	CAMBOU B, KLIBANOV A M, 1984. Preparative production of optically active esters and alcohols using esterase-catalyzed stereospecific transesterification in organic media[J]. Journal of the American Chemical Society, 106(9): 2687-2692.
[6]	CHANG LEI, OUYANG LIMING, XU YI, et al, 2010. Highly enantioselective hydrolysis of phenyl-1,2-ethanediol cyclic carbonates by newly isolated Bacillus sp. ECU0015[J]. Journal of Molecular Catalysis B: Enzymatic, 66(1/2): 95-100.
[7]	CHEN C S, FUJIMOTO Y, GIRDAUKAS G, et al, 1982. Quantitative analyses of biochemical kinetic resolutions of enantiomers[J]. Journal of the American Chemical Society, 104(25): 7294-7299.
[8]	FAN YONGXIAN, XIE ZHANGMING, ZHANG HUAWEI, et al, 2011. Kinetic resolution of both 1-phenylethanol enantiomers produced by hydrolysis of 1-phenylethyl acetate with Candida antarctica lipase B in different solvent systems[J]. Kinetics and Catalysis, 52(5): 686-690.
[9]	FERNÁNDEZ-ÁLVARO E, KOURIST R, WINTER J, et al, 2010. Enantioselective kinetic resolution of phenylalkyl carboxylic acids using metagenome-derived esterases[J]. Microbial Biotechnology, 3(1): 59-64.
[10]	FILLAT A, ROMEA P, PASTOR F I J, et al, 2015. Kinetic resolution of esters from secondary and tertiary benzylic propargylic alcohols by an improved esterase-variant from Bacillus sp. BP-7[J]. Catalysis Today, 255: 16-20.
[11]	GIRI A, DHINGRA V, GIRI C C, et al, 2001. Biotransformations using plant cells, organ cultures and enzyme systems: current trends and future prospects[J]. Biotechnology Advances, 19(3): 175-199.
[12]	KITAGUCHI H, FITZPATRICK P A, HUBER J E, et al, 1989. Enzymic resolution of racemic amines: crucial role of the solvent[J]. Journal of the American Chemical Society, 111(8): 3094-3095.
[13]	LAANE C, BOEREN S, VOS K, et al, 1987. Rules for optimization of biocatalysis in organic solvents[J]. Biotechnology Bioengineering, 30(1): 81-87.
[14]	NOJIRI M, TAOKA N, YASOHARA Y, 2014. Characterization of an enantioselective amidase from Cupriavidus sp. KNK-J915 (FERM BP-10739) useful for enzymatic resolution of racemic 3-piperidinecarboxamide[J]. Journal of Molecular Catalysis B: Enzymatic, 109: 136-142.
[15]	PANDA T, GOWRISHANKAR B S, 2005. Production and applications of esterases[J]. Applied Microbiology and Biotechnology, 67(2): 160-169.
[16]	SALAMEH M, WIEGEL J, 2007. Lipases from extremophiles and potential for industrial applications[J]. Advances in Applied Microbiology, 61: 253-283.
[17]	SMITH M E, BANERJEE S, SHI YONGLIANG, et al, 2012. Investigation of the cosolvent effect on six isoenzymes of PLE in the enantioselective hydrolysis of selected α,α-disubstituted malonate esters[J]. Chemcatchem, 4(4): 472-475.
[18]	TANG XIAOLING, LIU JI, WANG BO, et al, 2011. Cloning, screening and characterization of enantioselective ester hydrolases from Escherichia coli K-12[J]. World Journal of Microbiology & Biotechnology, 27(1): 129-136.
[19]	THÖRN C, GUSTAFSSON H, OLSSON L, 2011. Immobilization of feruloyl esterases in mesoporous materials leads to improved transesterification yield[J]. Journal of Molecular Catalysis B: Enzymatic, 72(1/2): 57-64.
[20]	TIAN XIN, ZHENG GAOWEI, LI CHUNXIU, et al, 2011. Enantioselective production of (S)-1-phenyl-1,2-ethanediol from dicarboxyesters by recombinant Bacillus subtilis esterase[J]. Journal of Molecular Catalysis B: Enzymatic, 73(1/2/3/4): 80-84.
[21]	WEBER N, GORWA-GRAUSLUND M, CARLQUIST M, 2014. Engineered baker’s yeast as whole-cell biocatalyst for one-pot stereo-selective conversion of amines to alcohols[J]. Microbial Cell Factories, 13(1): 118-127.
[22]	ZHENG JIANYONG, WANG YUQIANG, LUO WEIFENG, et al, 2015. Biocatalytic resolution of Boc-DL-alanine methyl ester by a newly isolated Bacillus amyloliquefaciens WZZ002[J]. Catalysis Communications, 60: 134-137.
[23]	ZHENG RENCHAO, ZHENG YUGUO, LI AIPENG, et al, 2014. Enantioselective synthesis of (S)-3-cyano-5-methylhexanoic acid by a high DMSO concentration tolerable Arthrobacter sp. ZJB-09277[J]. Biochemical Engineering Journal, 83: 97-103.
[24]	ZOU SHUPING, ZHENG YUGAO, DU ERHONG, et al, 2014. Enhancement of (S)-2,3-dichloro-1-propanol production by recombinant whole-cell biocatalyst in n-heptane-aqueous biphasic system[J]. Journal of Biotechnology, 188: 42-47.

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

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

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