Synthesis of (R)-epichlorohydrin catalyzed by cross-linked cell aggregates of epoxide hydrolase

ZOU Shuping1 JIANG Zhentao1 WANG Zhicai1 LIU Zhiqiang1 ZHENG Yuguo1

(1.National and Local Joint Engineering Research Center for Chiral Biomanufacturing, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang, China 310014)

【Abstract】Polyethyleneimine (PEI) flocculation and glutaraldehyde (GA) cross-linking were used to prepare cross-linked cell aggregates (CLCAs) of whole cells of epoxide hydrolase. The effects of PEI concentration, GA concentration, and diatomite carrier dosage on the activity recovery of CLCAs were investigated. The results show that the optimal values of PEI concentration, GA concentration, and diatomite carrier dosage are 3% (vol), 1% (vol), and 6 g/L, respectively and the activity recovery reaches 88.4%. Using CLCAs as catalysts, racemic epichlorohydrin [(R,S)-ECH] as substrate, the synthesis of (R)-epichlorohydrin [(R)-ECH] in isooctane/phosphate buffer biphase system was investigated. The results show that under the condition of 3:7 volume ratio of isooctane to buffer, substrate concentration of 800 mmol/L, addition of 18 g/L CLCAs, buffer pH 8.0, temperature at 35 °C, the molar yield of (R)-ECH reaches 45.2%, with 99.1% ee. The operational stability of CLCAs in the biphase system was investigated, and the viability of nine batches reused remains basically unchanged, showing good operation stability.

【Keywords】 epoxide hydrolase; cross-linked cell aggregates; immobilization; biocatalysis; multiphase reaction; chiral epichlorohydrin;

【DOI】

【Funds】 National Natural Science Foundation of China (21406205, 21176224)

Download this article

(Translated by WANG YX)

    References

    [1] Wan N W, Liu Z Q, Xue F, et al. A one-step biocatalytic process for (S)-4-chloro-3-hydroxybutyronitrile using halohydrin dehalogenase: a chiral building block for atorvastatin [J].Chemcatchem, 2015, 7 (16): 2446–2450.

    [2] Kabat M M, Daniewski A R, Burger W. A convenient synthesis of R-(−)-carnitine from R-(−)-epichlorohydrin [J]. Tetrahedron: Asymmetry, 1997, 8 (16): 2663–2665.

    [3] van Leeuwen J G E, Wijma H J, Floor R J, et al. Directed evolution strategies for enantiocomplementary haloalkane dehalogenases: from chemical waste to enantiopure building blocks [J]. ChemBioChem, 2012, 13 (1): 137–148.

    [4] Qu Y Q. Production process of epoxy chloropropane and analysis of its domestic and overseas market [J]. Technology & Economics in Petrochemicals, 2015, 31 (4): 26–30 (in Chinese).

    [5] Zhang X J, Zheng Y G. Research and development on key enzymes for biosynthesis of chiral epichlorohydrin using glycerol [J]. Biotechnology & Business, 2017, 6: 65–72 (in Chinese).

    [6] Zhao Z P, Li M S, Zhang J Y, et al. New chiral catalytic membranes created by coupling UV-photografting with covalent immobilization of Salen-Co (Ⅲ) for hydrolytic kinetic resolution of racemic epichlorohydrin [J]. Ind. Eng. Chem. Res., 2012, 51 (28): 9531–9539.

    [7] Xue F, Liu Z Q, Zou S P, et al. A novel enantioselective epoxide hydrolase from Agromyces mediolanus ZJB120203: cloning, characterization and application [J]. Process Biochem., 2014, 49: 409–417.

    [8] Bala N, Chimni S S. Recent developments in the asymmetric hydrolytic ring opening of epoxides catalysed by microbial epoxide hydrolase [J]. Tetrahedron: Asymmetry, 2010, 21 (24): 2879–2898.

    [9] Kim H S, Lee J H, Park S, et al. Biocatalytic preparation of chiral epichlorohydrins using recombinant Pichia pastoris expressing epoxide hydrolase of Rhodotorula glutinis [J]. Biotechnol. Bioprocess Eng., 2004, 9 (1): 62–64.

    [10] Woo J H, Hwang Y O, Kang J H, et al. Enantioselective hydrolysis of racemic epichlorohydrin using an epoxide hydrolase from Novosphingobium aromaticivorans [J]. J. Biosci. Bioeng., 2010, 110 (3): 295–297.

    [11] Xu H H, Chen Y Y, Hu Z C, et al. Screening, identification, and enzyme-producing conditions of chiral resolution of epichlorohydrins [J]. J. Microbiol., 2009, 29 (2): 33–38 (in Chinese).

    [12] Zajkoska P, Rebros M, Rosenberg M. Biocatalysis with immobilized Escherichia coli [J]. Appl. Microbiol. Biotechnol., 2013, 97 (4): 1441–1455.

    [13] Dicosimo R, Mcauliffe J, Poulose A J, et al. Industrial use of immobilized enzymes [J]. Chem. Soc. Rev., 2013, 42 (15): 6437–6474.

    [14] Maritz J, Krieg H M, Yeates C A, et al. Calcium alginate entrapment of the yeast Rhodosporidium toruloides for the kinetic resolution of 1,2-epoxyoctane [J]. Biotechnol. Lett., 2003, 25 (20): 1775–1781.

    [15] Yildirim D, Tukel S S, Alptekin O, et al. Immobilized Aspergillus niger epoxide hydrolases: cost-effective biocatalysts for the preparation of enantiopure styrene oxide, propylene oxide and epichlorohydrin [J]. J. Mol. Catal. B—Enzym., 2013, 88: 84–90.

    [16] Kroutil W, Orru R V A, Faber K. Stabilization of Nocardia EH1 epoxide hydrolase by immobilization [J]. Biotechnol. Lett., 1998, 20 (4): 373–377.

    [17] Kim Y H, Lee I, Choi S H, et al. Nanoimmobilization of marine epoxide hydrolase of Mugil cephalus for repetitive enantioselective resolution of racemic styrene oxide in aqueous buffer [J]. J.Nanosci. Nanotechnol., 2013, 13 (3): 2266–2271.

    [18] Becka S, Skrob F, Plhackova K, et al. Cross-linked cell aggregates of Trigonopsis variabilis: D-amino acid oxidase catalyst for oxidation of cephalosporin C [J]. Biotechnol. Lett., 2003, 25 (3): 227–233.

    [19] Zou S P, Huang J W, Xue Y P, et al. Highly efficient production of 1-cyanocyclohexaneacetic acid by cross-linked cell aggregates (CLCAs) of recombinant E. coli harboring nitrilase gene [J].Process Biochem., 2018, 65: 93–99.

    [20] Jin H X, Liu Z Q, Hu Z C, et al. Production of (R)-epichlorohydrin from 1,3-dichloro-2-propanol by two-step biocatalysis using haloalcohol dehalogenase and epoxide hydrolase in two-phase system [J]. Biochem. Eng. J., 2013, 74: 1–7.

    [21] Zou S P, Zheng Y G, Wu Q, et al. Enhanced catalytic efficiency and enantioselectivity of epoxide hydrolase from Agrobacterium radiobacter AD1 by iterative saturation mutagenesis for (R)-epichlorohydrin synthesis [J]. Appl. Microbiol. Biotechnol., 2018, 102 (2): 733–742.

    [22] Chen J, Zheng Y G, Shen Y C. Biosynthesis of p-methoxyphenylacetic acid from p-methoxyphenylacetonitrile by immobilized Bacillus subtilis ZJB-063 [J]. Process Biochem., 2008, 43 (9): 978–983.

    [23] Shen M, Zheng Y G, Liu Z Q, et al. Production of acrylic acid from acrylonitrile by immobilization of Arthrobacter nitroguajacolicus ZJUTB06-99 [J]. J. Microbiol. Biotechnol., 2009, 19 (6): 582–587.

    [24] Zou S P, Yan H W, Hu Z C, et al. Synthesis of (R)-epichlorohydrin catalyzed by immobilized recombinant Escherichia coli cells [J]. Modern Chemical Industry, 2013, 33 (7): 55–59 (in Chinese).

    [25] Wang Y J, Chen X P, Shen W, et al. Chiral diol t-butyl 6-cyano-(3R,5R)-dihydroxylhexanoate synthesis catalyzed by immobilized cells of carbonyl reductase and glucose dehydrogenase coexpression E. coli [J]. Biochem. Eng. J., 2017, 128: 54–62.

    [26] Zhang Z J, Pan J, Li C X, et al. Efficient production of (R)-(−)-mandelic acid using glutaraldehyde cross-linked Escherichia coli cells expressing Alcaligenes sp. nitrilase [J]. Bioprocess. Biosyst.Eng., 2014, 37 (7): 1241–1248.

    [27] Deng H, Chen S, Chen J, et al. Immobilization of cells producing glucose isomerases [J]. Food Science, 2013, 34 (9): 164–169 (in Chinese).

    [28] Chen W, Lou W, Wang X, et al. Asymmetric hydrolysis of styrene oxide catalyzed by mung bean epoxide hydrolase in organic solvent/buffer biphasic system [J]. Chin. J. Catal., 2011, 32 (9): 1557–1563.

    [29] Fuhshuku K, Asano Y. Synthesis of (R)-beta-nitro alcohols catalyzed by R-selective hydroxynitrile lyase from Arabidopsis thaliana in the aqueous-organic biphasic system [J]. J.Biotechnol., 2011, 153 (3/4): 153–159.

    [30] Li A T, Zhang J D, Yu H L, et al. Significantly improved asymmetric oxidation of sulfide with resting cells of Rhodococcus sp in a biphasic system [J]. Process Biochem., 2011, 46 (3): 689–694.

    [31] Zou S, Yan H, Hu Z, et al. Enzymatic resolution of epichlorohydrin catalyzed by whole cells in an organic solvent/buffer biphasic system [J]. Chin. J. Catal., 2013, 34 (7): 1339–1347.

    [32] Gong P F, Xu J H. Bio-resolution of a chiral epoxide using whole cells of Bacillus megaterium ECU1001 in a biphasic system [J].Enzyme Microb. Technol., 2005, 36 (2/3): 252–257.

    [33] Xue Y P, Zhong H J, Zou S P, et al. Efficient chemoenzymatic synthesis of gabapentin by control of immobilized biocatalyst activity in a stirred bioreactor [J]. Biochem. Eng. J., 2017, 125: 190–195.

    [34] Liang Y, Jiao S, Wang M, et al. Overexpression of epoxide hydrolase in Rhodococcus ruber with high robustness for the synthesis of chiral epichlorohydrin [J]. Process Biochem., 2019, 79: 49–56.

    [35] Zou S P, Wang Z C, Qin C, et al. Covalent immobilization of Agrobacterium radiobacter epoxide hydrolase on ethylenediamine functionalised epoxy supports for biocatalytical synthesis of (R)-epichlorohydrin [J]. Biotechnol. Lett., 2016, 38 (9): 1579–1585.

This Article

ISSN:0438-1157

CN: 11-1946/TQ

Vol 71, No. 09, Pages 4238-4245

September 2020

Downloads:0

Share
Article Outline

Abstract

  • Introduction
  • 1 Experimental materials and methods
  • 2 Results and discussion
  • 3 Conclusion
  • References