深海来源微生物乙酰酯酶的酶学性质鉴定及拆分制备D-乳酸甲酯

doi:10.11978/2017106

Abstract

Abstract:

D-lactic acid and its esters are important chiral drug intermediates and chemicals. An acetyl esterase, Bae02030, from Bacillus sp. SCSIO15029 isolated from the deep sea of the South China Sea was cloned, expressed and functionally characterized. The optimum pH and temperature of acetyl esterase Bae02030 were 8.5 and 35°C, respectively. Bae02030 exhibited excellent resistance to most organic solvents and surfactants tested. Bae02030 could generate optically pure D-methyl lactate through resolution of racemic methyl lactate. After process optimization of the enzymatic resolution reactions, the addition of 60% heptane improved the enantio-selectivity of Bae02030. The enantiomeric excess of prepared D-methyl lactate was over 99% and conversion rate reached 56%. Acetyl esterase Bae02030 identified from deep-sea microorganism, as a biocatalyst, possesses great potential in the production of chiral drug intermediates in industry.

Key words: deep-sea microorganisms, acetyl esterase, biocatalysis, D-methyl lactate, chiral resolution

CLC Number:

P735.51

Jinlong HUANG, Jifu ZHANG, Jieying HU, Jianliu GUAN, Yun ZHANG, Aijun SUN, Yunfeng HU. Characterization of one deep-sea derived microbial acetyl esterase and its utilization in the preparation of D-methyl lactate through kinetic resolution[J].Journal of Tropical Oceanography, 2018, 37(4): 38-44.

Figures/Tables 8

Fig. 1

Fig. 2

Fig. 3

Tab. 1

Tab. 2

Fig. 4

Fig. 5

Tab. 3

References 19

1	周明芳, 2014. (R)-2-[4-(6-氯-2-苯并噁唑氧基)苯氧基丙酸的合成工艺研究[D]. 南京: 南京理工大学.
	ZHOU MINGFANG, 2014. Study on synthesis of (R)-2-[4-(6-Chlor-2- benzoxazolyloxy)-phenoxy]-propionic acid[D]. Nanjing: Nanjing University of Science & Technology (in Chinese with English abstract).
2	AQAR D Y, RAHMANIAN N, MUJTABA I M, 2016. Methyl lactate synthesis using batch reactive distillation: Operational challenges and strategy for enhanced performance[J]. Separation and Purification Technology, 158: 193-203.
3	CHANG P C, TSAI W S, TSAI C L, et al, 2004. Cloning and characterization of two thermostable xylanases from an alkaliphilic Bacillus firmus[J]. Biochemical and Biophysical Research Communications, 319(3): 1017-1025.
4	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.
5	FAULDS C B, PÉREZ-BOADA M, MARTÍNEZ Á T, 2011. Influence of organic co-solvents on the activity and substrate specificity of feruloyl esterases[J]. Bioresource Technology, 102(8): 4962-4967.
6	HUANG JINLONG, ZHANG YUN, HU YUNFENG, 2016. Functional characterization of a marine Bacillus esterase and its utilization in the stereo-selective production of D-methyl lactate[J]. Applied Biochemistry and Biotechnology, 180(8): 1467-1481.
7	HUY N C, THIYAGARAJAN S, KIM D H, et al, 2013. Cloning and characterization of a novel bifunctional acetyl xylan esterase with carbohydrate binding module from Phanerochaete chrysosporium[J]. Journal of Bioscience and Bioengineering, 115(5): 507-513.
8	KWON M, SONG J, PARK H S, et al, 2016. Characterization of heterologously expressed acetyl xylan esterase1 isolated from the anaerobic rumen fungus Neocallimastix frontalis PMA02[J]. Asian-Australas Journal of Animal Sciences, 29(11): 1576-1584.
9	MILLAR R, RAHMANPOUR R, YUAN E W J, et al, 2016. Esterase EstK from Pseudomonas putida mt-2: An enantioselective acetylesterase with activity for deacetylation of xylan and poly (vinylacetate)[J]. Biotechnology and Applied Biochemistry, 64(6): 803-809.
10	MØLGAARD A, KAUPPINEN S, LARSEN S, 2000. Rhamnogalacturonan acetylesterase elucidates the structure and function of a new family of hydrolases[J]. Structure, 8(4): 373-383.
11	NAVARRO-FERNÁNDEZ J, MARTÍNEZ-MARTÍNEZ I, MONTORO-GARCÍA S, et al, 2008. Characterization of a new rhamnogalacturonan acetyl esterase from Bacillus halodurans C-125 with a new putative carbohydrate binding domain[J]. Journal of Bacteriology, 190(4): 1375-1382.
12	REDDY C R, LATHA B, WARUDIKAR K, et al, 2016. Total synthesis of a piperidine alkaloid, microcosamine A[J]. Organic & Biomolecular Chemistry, 14(1): 251-258.
13	REMOROZA C, WAGENKNECHT M, GU F, et al, 2014. A Bacillus licheniformis pectin acetylesterase is specific for homogalacturonans acetylated at O-3[J]. Carbohydrate Polymers, 107: 85-93.
14	RICHARD G, NOTT K, NICKS F, et al, 2013. Use of lipases for the kinetic resolution of lactic acid esters in heptane or in a solvent free system[J]. Journal of Molecular Catalysis B: Enzymatic, 97: 289-296.
15	TIAN QIANQIAN, SONG PING, JIANG LING, et al, 2014. A novel cephalosporin deacetylating acetyl xylan esterase from Bacillus subtilis with high activity toward cephalosporin C and 7-aminocephalosporanic acid[J]. Applied Microbiology and Biotechnology, 98(5): 2081-2089.
16	TONG XIAOXUE, LANGE L, GRELL M N, et al, 2015. Hydrolysis of wheat arabinoxylan by two acetyl xylan esterases from Chaetomium thermophilum[J]. Applied Biochemistry and Biotechnology, 175(2): 1139-1152.
17	WANG YILONG, ZHANG YUN, HU YUNFENG, 2016. Functional characterization of a robust marine microbial esterase and its utilization in the stereo-selective preparation of ethyl (S)-3-hydroxybutyrate[J]. Applied Biochemistry and Biotechnology, 180(6): 1196-1212.
18	YAMANE H, SASAI K, 2003. Effect of the addition of poly (D-lactic acid) on the thermal property of poly (L-lactic acid)[J]. Polymer, 44(8): 2569-2575.
19	ZHOU WEI, LIU XIAOHUI, YE LI, et al, 2014. The biotransformation of astragalosides by a novel acetyl esterase from Absidia corymbifera AS2[J]. Process Biochemistry, 49(9): 1464-1471.

pH	对映体过量值/%	转化率/%	温度/°C	对映体过量值/%	转化率/%
7.5	35.85 ± 0.31	53.73 ± 0.25	30	71.17 ± 0.44	55.29 ± 0.40
8.0	48.13 ± 0.38	55.0 ± 0.30	35	76.33 ± 0.28	56.42 ± 0.18
8.5	66.05 ± 0.62	56.23 ± 0.29	37	81.17 ± 0.35	57.86 ± 0.51
9.0	76.45 ± 0.53	56.96 ± 0.52	40	83.29 ± 0.53	58.47 ± 0.32
9.5	65.21 ± 0.33	58.21 ± 0.41	45	79.87 ± 0.71	60.08 ± 0.33
10.0	63.79 ± 0.19	59.31 ± 0.38	50	64.79 ± 0.36	59.57 ± 0.59

	lgP	对映体过量值/%	转化率/%
对照	0	89.23 ± 0.42	66.60 ± 0.55
正己烷	3.94	89.15 ± 0.27	66.76 ± 0.32
环己烷	3.39	90.27 ± 0.46	66.32 ± 0.42
正庚烷	4.47	90.53 ± 0.52	61.34 ± 0.20
甲醇	-0.72	52.89 ± 0.63	54.97 ± 0.46
甲苯	2.68	84.23 ± 0.35	64.60 ± 0.34
丙酮	-0.16	50.19 ± 0.30	45.88 ± 0.09
DMSO	-1.35	88.89 ± 0.73	61.94 ± 0.24
曲拉通	0	88.23 ± 0.33	67.30 ± 0.22
吐温-20	0	90.53 ± 0.25	69.80 ± 0.88
吐温-80	0	91.33 ± 0.41	66.42 ± 0.27
STPP	0	90.15 ± 0.95	67.57 ± 1.04

底物浓度 /(mmol·L^-1)	时间/h	对映体过量值/%	转化率/%	D-乳酸甲酯产率 /(mmol·L^-1·h^-1)
12.5	2	> 99.5	56.03 ± 0.43	2.75 ± 0.03
25.0	4	> 99.5	61.43 ± 0.26	2.41 ± 0.03
37.5	6	> 99.5	63.35 ± 0.35	2.29 ± 0.04
50.0	6	87.07 ± 0.31	54.58 ± 0.17	1.78 ± 0.01

[1]	LI Cun, CUI Linqing, YANG Hongqiang, LONG Lijuan, TIAN Xinpeng. Diversity of cultured bacteria isolated from three coral reef sediments in South China Sea [J]. Journal of Tropical Oceanography, 2022, 41(2): 149-158.
[2]	YU Fei, JIN Xingkun, LEI Tianying, CAO Haihang, CHEN Qianghui, YANG Yaofan, LI Jiahang, ZHAO Zhe. Heterologous expression and enzymatic characterization of marine Vibrio astriarenae-derived β-Agarase gene vas1-1339 [J]. Journal of Tropical Oceanography, 2022, 41(2): 170-176.
[3]	ZHANG Lele, ZOU Qiang, TIAN Yanan, LÜ Chunhui, ZHENG Shiyi, JIANG Guangjun, GAO Longkun, HOU Yuping, WANG Kai. Dysbiosis of both structure and function of intestinal microbiota in lined seahorses (Hippocampus erectus) as response to Edwardsiella tarda infection [J]. Journal of Tropical Oceanography, 2022, 41(2): 177-188.
[4]	LIU Wei, GUO Haipeng, DONG Pengsheng, YAN Mengchen, ZHANG Demin. Draft genome sequence and comparative genome analysis of Alliroseovarius sp. Z3 [J]. Journal of Tropical Oceanography, 2022, 41(1): 52-61.
[5]	MAO Yingjin, GAO Beile. Comparative genomic analysis of the distribution and evolution of quorum sensing pathways in the Vibrio genus [J]. Journal of Tropical Oceanography, 2022, 41(1): 28-41.
[6]	LU Chunju, LU Meilin, LIU Xinming, LIU Yonghong, GAO Chenghai, XU Xinya. Diversity and anti-bacteria activity of the gorgonian derived fungi from Weizhou Island of Guangxi Province^* [J]. Journal of Tropical Oceanography, 2021, 40(5): 45-52.
[7]	LIU Bingxin, WEI Xia, XIAO Xiji, ZHANG Qi, ZHANG Cuixian. Research on benzaldehydes from the soft coral-associated symbiotic fungus Aspergillus sp. EGF15-0-3 [J]. Journal of Tropical Oceanography, 2021, 40(4): 63-69.
[8]	LI Yanqun, CHEN Rouwen, LIN Zonghao, TIAN Xinpeng, YIN Hao. Screening and identification of a quorum sensing inhibitory actinomycetes derived from marine sediments [J]. Journal of Tropical Oceanography, 2021, 40(1): 75-81.
[9]	HAN Minmin, LI Mi, LIU Xinming, LIU Yonghong, LONG Chao, ZHONG Zhenguo, YI Xiangxi, GAO Chenghai. Studies on bacterial diversity in coral reef sediments in Khai Island and Pathiu Island and bacterial crude extract retards aging activity of Caenorhabditis elegans* [J]. Journal of Tropical Oceanography, 2020, 39(5): 19-29.
[10]	Xiangfu LI, Jie XU, Zhen SHI, Ruihuan LI. Spatial and temporal variation in heterotrophic bacteria and their regulators in the Pearl River Estuary [J]. Journal of Tropical Oceanography, 2018, 37(1): 27-36.
[11]	WANG Zhao-kai, YI Zhi-wei, LIU Yang, CHAN Zhu-hua, ZENG Run-ying. Isolation, identification and characterization of an esterase-producing marine moderately halophilic strain Halomonas sp.LYG1-1 [J]. Journal of Tropical Oceanography, 2014, 33(1): 69-73.
[12]	ZHANG Ling, ZHANG Qing-bo, CHEN Yu-chan, LI Hao-hua, WANG Lei, ZHANG Wei-min. Preliminary identification of 18 marine fungal strains from the South China Sea and screening of their fermentation products for cytotoxic and antibacterial activities [J]. Journal of Tropical Oceanography, 2013, 32(3): 47-51.

Characterization of one deep-sea derived microbial acetyl esterase and its utilization in the preparation of D-methyl lactate through kinetic resolution

RichHTML

PDF (PC)

Like

Knowledge

Cited

Abstract

Cite this article

share this article

Figures/Tables 8

References 19

Related Articles 12

Metrics

Comments

Recommended 0