A Recyclable Poly(ionic liquid)s Enzyme Reactor for Highly Efficient Protein Digestion

Document Type: Research Article

Authors

College of Chemical Engineering and Materials Science, Tianjin University of Science and Technology, Tianjin 300457, P.R. CHINA

Abstract

One of the most significant tasks for proteomic research and industrial applications, is the preparation of recyclable enzyme reactor. Herein, a novel recyclable enzyme reactor has been developed based on monodispersed spherical poly(quaternary ammonium ionic liquid)s particles immobilized trypsin. A new quaternary ammonium ionic liquids functional monomer was first synthesized. The ionic liquids functional monomer was then copolymerized with ethylene glycol dimethacrylate by precipitation polymerization. The resultant monodispersed spherical particle showed a large surface area (231 m2/g) and high binding capacity for trypsin (200 mg/g) due to the large surface area and strong interaction. The polymer microsphere loaded trypsin was used as an enzyme reactor for the digestion of standard protein, semi-complex samples and skim milk, respectively. The results indicated that the enzyme reactor exhibited highly efficient protein digestion and excellent stability. The digestion time of the present ionic liquids enzyme reactor for the digestion of protein, the solution could be reduced to even 5 min. The ionic liquids enzyme reactor showed excellent reusability and could be reused for more than four times. When it was kept at 4 °C for 12 d, and used for skim milk digestion, the obtained MALDI-TOF score could also reach 88 with 29 matched peptides.

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Main Subjects


[1] Rogers R.D., Voth G.A., Ionic liquids, Acc. Chem. Res., 40:1077-1078 (2007).

[2] Wang J.L., Yao H.W., Nie Y., Zhang X.P., Li J.W., Synthesis and Characterization of the Iron-Containing Magnetic Ionic Liquids, J. Mol. Liq., 169: 152-155 (2012).

[3] Mao G.X., Zhu A.F., The Aggregation Behavior of Short Chain Hydrophilic Ionic Liquids in Aqueous Solutions, Iran. J. Chem. & Chem. Eng. (IJCCE), 32: 77-82 (2013).

[4] Hallett J.P., Welton T., Room-Temperature Ionic Liquids: Solvents for Synthesis and Catalysis 2, Chem. Rev., 111:3508–3576 (2011).

[5] Wasserscheid P., Keim W., Ionic Liquids-New Solutions for Transition Metal Catalysis, Angew Chem. Int. Edit., 39:3772–3789 (2000).

[6] Wang H., Gurau G., Rogers R.D., Ionic Liquid Processing of Cellulose, Chem. Soc. Rev., 41:1519-1537 (2012).

[7] Walsh D.A., Lovelock K.R., Licence P., Ultramicroelectrode Voltammetry and Scanning Electrochemical Microscopy in Room-Temperature Ionic Liquid Electrolytes, Chem. Soc. Rev., 39: 4185–4194 (2010).

[8] Dupont J., Scholten J.D., On the Structural and Surface Properties of Transition-Metal Nanoparticles in Ionic Liquids, Chem. Soc. Rev., 39: 1780–1804 (2010).

[9] Bideau J.L., Viau L., Vioux A., Ionogels, Ionic Liquid Based Hybrid Materials, Chem. Soc. Rev., 40: 907–925 (2011).

[10] Kragl U., Eckstein M., Kaftzik N., Enzyme Catalysis in Ionic Liquids, Curr. Opin. Chem. Biol., 13: 565-571 (2002).

[11] Gao W.W., Zhang F. X., Zhang G.X., Zhou C.H., Key Factors Affecting the Activity and Stability of Enzymes in Ionic Liquids and Novel Applications in Biocatalysis, Biochem. Eng. J., 99: 67-84 (2015).

[12] Barron C.C., Sponagle B.J.D., Arivalagan P., Cunha G.B.D., Optimization of Oligomeric Enzyme Activity in Ionic Liquids Using Rhodotorula Glutinis Yeast Phenylalanine Ammonia Lyase, Enzyme Microbial Tech., 96: 151-156 (2017). 

[13] Ou G.N., He B.Y., Halling P., Ionization Basis for Activation of Enzymes Soluble in Ionic Liquids, Biochim. Biophys. Acta, 1860: 1404-1408 (2016).

[14] Solhtalab M., Karbalaei-Heidari H. R., Absalan G., Tuning of Hydrophilic Ionic Liquids Concentration: A Way to Prevent Enzyme Instability, J. Molecular Catalysis B: Enzymatic, 122: 125-130 (2015).

[15] Misuk V., Breuch D., Löwe H., Paramagnetic Ionic Liquids as “Liquid Fixed-Bed” Catalysts in Flow Application, Chem. Eng. J., 173: 536-540 (2011).

[17] Sahiner N., Demir S., Yildiz S., Magnetic Colloidal Polymeric Ionic Liquid Synthesis and Use in Hydrogen Production, Colloid. Surface. A, 449: 87-95 (2014).

[18] Eftekhari A., Saito T., Synthesis and Properties of Polymerized Ionic Liquids, Eur. Poly. J., 90:245-272 (2017).

[19] Men Y.J., Kuzmicz D., Yuan JY., Poly(ionic liquid) Colloidal Particles, Current Opinion Colloid & Inter. Sci., 19: 76-83 (2014).

[20] Paino M.Á., Bonilla A.M., Fabal F.L., Garcés J.L., Heuts J.P.A., García M.F., Effect of Glycounits on the Antimicrobial Properties and Toxicity behavior of Polymers Based on Quaternized DMAEMA, Biomacromolecules, 16: 295-303 (2015).

[21] Rantwijk F.V., Sheldon R. A., Biocatalysis in Ionic Liquids, Chem. Rev., 107:2757-2785 (2007).

[23] Qiao J., Kim J.Y., Wang Y.Y., Qi L., Wang F.Y., Moon M.H., Trypsin Immobilization in Ordered Porous Polymer Membranes for Effective Protein Digestion, Anal. Chim. Acta., 906: 56-64 (2016).

[24] Switzar L., Giera M., Niessen W.M.A., Protein Digestion: an Overview of the Available Techniques and Recent Developments, J. Proteome. Res., 12: 1067-1077 (2013).

[27] Wang H.P., Jiao F.L., Gao F.Y., Zhao X.Y., Zhao Y., Shen Y.H., Covalent Organic Framework-Coated Magnetic Graphene as a Novel Support for Trypsin Immobilization, Anal. Bioanal. Chem., 409: 2179-2187 (2017).

[28] Fan C., Shi Z., Pan Y., Song Z., Zhang W., Zhao X., Dual Matrix-Based Immobilized Trypsin for Complementary Proteolytic Digestion and Fast Proteomics Analysis with Higher Protein Sequence Coverage, Anal Chem., 86(3): 1452-1458 (2014).

[29] Karamatollah R., Feral T., Changes in Enzyme Efficiency During Lipase-Catalyzed Hydrolysis of Canola Oil in a Supercritical Bioreactor, Iran. J. Chem. Chem. Eng. (IJCCE), 25(4): 25-35 (2006).

[30] Jiang B., Yang K., Zhang L., Liang Z., Peng X., Zhang Y., Dendrimer-Grafted Graphene Oxide Nanosheets as Novel Support for Trypsin Immobilization to Achieve Fast on-Plate Digestion of Proteins, Talanta, 122: 278-284 (2014).

[31] Cheng G., Zheng S.Y., Construction of a High-Performance Magnetic Enzyme Nanosystem for Rapid Tryptic Digestion, Scientific Reports, 4:6947 (2014).

DOI: 10.1038/srep06947

[32] Shi C., Deng C., Li Y., Zhang X., Yang P., Hydrophilic Polydopamine-Coated Magnetic Graphene Nanocomposites for Highly Efficient Tryptic Immobilization, Proteomics, 14: 1457-1463 (2014).

[33] Cao Y., Wen L.Y., Svec F., Tan T.W., Lv Y.Q., Magnetic AuNP@Fe3O4 Nanoparticles as Reusable Carriers for Reversible Enzyme Immobilization, Chem. Eng. J., 286: 272-281 (2016).

[34] Lin Z.A., Xiao Y., Wang L., Yin Y.Q., Zheng J.N., Yang H.H., Chen, G.N., Facile Synthesis of Enzyme–Inorganic Hybrid Nanoflowers and Their Application as an Immobilized Trypsin Reactor for Highly Efficient Protein Digestion, RSC Adv., 4: 13888-13891 (2014).

[35] Haupt K., Bueno S.M.A., Vijayalakshmi M.A., Interaction of Human Immunoglobulin G with l-Histidine Immobilized onto Poly(ethylene vinyl alcohol) Hollow-Fiber Membranes, J. Chromatogr. B, 674: 13-21 (1995).

[36] Jin L., He D., Li Z., Wei M., Protein Adsorption on Gold Nanoparticles Supported by a Layered Double Hydroxide, Mat. Lett., 77: 67-70 (2012).

[37] Bellezza F., Alberani A., Posati T., Tarpani L., Latterini L., Cipiciani A., Protein Interactions with Nanosized Hydrotalcites of Different Composition, J. Inorg. Biochem., 106: 134-142 (2012).

[38] Shi C., Deng C., Li Y., Zhang X., Yang P., Hydrophilic Polydopamine-Coated Magnetic Graphene Nanocomposites for Highly Efficient Tryptic Immobilization, Proteomics, 14: 1457-1463 (2014).

[39] Cao Y., Wen, L.Y., Svec F., Tan T.W., Lv Y.Q., Magnetic AuNP@Fe3O4 Nanoparticles as Reusable Carriers for Reversible Enzyme Immobilization, Chem. Eng. J., 285(15): 272-281 (2016).

[40] Ruan G.H., Wei M.P., Chen Z.Y., Su R.H., Du F.Y., Zheng Y.J., Novel Regenerative Large-Volume Immobilized Enzyme Reactor:Preparation, Characterization and Application, J. Chromatogr. B, 967: 13-20 (2014).